Volume XIX Issue No. 01 01 June 2020 Pages: 148 - Agrobios ...

9 779727 027003

ISSN 972-7027X

Volume XIX Issue No. 01 01 June 2020 Pages: 148

AGROBIOS NEWSLETTER Publishing Date || 01 June 2020

VOL. NO. XIX, ISSUE NO. 01 1

CONTENTS

BIOTECHNOLOGY

1. A New Pillar for Cancer Treatment: Reverse Immunology (Nobel Prize Winning Topic-2018) 5Ramachandra Anantapur and Roshni M

2. Micropropagation: A Technique for Rapid Propagation of Disease-Free Plants 7Minakshi Onkar Birajdar

3. Recombineering: A Lambda Red Based Tool for Genetic Engineering 8*Suresh H. Antre and Sunil Subramanya A. E.

4. Transcriptomics: What and What for? 10Nusrat Perveen and Hidayatullah Mir

5. Combined Approach of Morphological and Molecular Diagnosis of Diseases in Plants 11Bansuli

6. Abiotic and Biotic Stress Signal Cross Talking in Plants 12Jyoti Prakash Sahoo and Upasana Mohapatra

7. Cisgenesis: Can it be an Alternative to Transgenesis? 14Upasana Mohapatra and Jyoti Prakash Sahoo

8. IdentificationandDifferentiationofCropsthroughDNA Barcoding 16S. G. Magar and V. G. Magar

MICROBIOLOGY

9. Plastic Lovers: A Green thought for Plastics 17Ms. Aswathy, J. C. and Dr. Shalini Pillai, P.

BIOCHEMISTRY

10. First and Second Genome of DNA Sequencing 19Payal Chakraborty and Binny Sharma

NANOTECHNOLOGY

11. Nanobiosensors: An Emerging Technology in Agriculture 20Mohammed Nisab C. P.

12. Role of Myconanoparticles in Phytopathogens Management 22Arjunsinh Rathava, Puja Pandey and Ranganath Swamy

AGRONOMY

13. Bio-Fortification:ANovelApproachtoTackleHidden Hunger and Malnutrition 23Manjanagouda S. Sannagoudar, Avijit Ghosh, Keerthi, M. C. and Hanamant M Halli

14. Controlled Release Fertilizer (CRF): New Approach for Nutrient Management 25Dr. Vijay Kumar Didal, Dr. Brijbhooshan, Dr. Shalini, Dr. Krishna Chaitanya and Dr. Rajendragouda Patil1

JUNE, 2020 / VOLUME XIX / ISSUE NO. 01

CHIEF EDITORDr. S. S. Purohit

ASSOCIATE EDITORDr. P. Balasubramaniyan (Madurai)

Dr. Tanuja Singh (Patna) Dr. Ashok Agrawal (Mathura)

Dr. H. P. Sharma (Ranchi) Dr. N. Kachhawha (Jaipur)

Dr. Anil Kumar (Patna)

EDITORIAL OFFICEAgro House, Behind Nasrani CinemaChopasani Road, Jodhpur - 342 003

Phone: +91-291-2643993E-mail: [email protected];

[email protected]: www.agrobiosonline.com

TYPESETTINGYashee Computers, Jodhpur

PRINTED BYManish Kumar, Chopra Offset, Jodhpur

PUBLISHED BYDr. Updesh Purohit, for Agrobios (India),

Behind Nasrani, Cinema, Chopasani Road, Jodhpur

RNI NO.: RAJENG/2002/8649ISSN:0972-7027

DISCLAIMER: The views expressed by the authors do not necessarily represent those of editorial board or publishers.

Although every care has been taken to avoid errors or omission, this magazine is being published on the condition

and undertaking that all the information given in this magazine is merely for reference and must not be taken as having authority of or binding in any way on the authors, editors and publishers who do not owe any responsibility

for any damage or loss to any person, for the result of any action taken on the basis of this work. The Publishers

shall be obliged if mistakes brought to their notice.

SUBSCRIPTION RATESSINGLE COPY: ` 85.00

ANNUAL INDIVIDUAL: ` 1000.00ANNUAL INSTITUTIONAL: ` 2000.00

© The articles published in Agrobios Newsletter is subject to copy right of the publisher. No material can be reproduced without prior permission of the publisher.Issues of “Agrobios Newsletter” are mailed by ordinary post at Subscriber’s risk and our responsibility ceases once we hand over the magazine to post office.

NOTE: “Agrobios Newsletter” does not accept unsolicited manuscripts and material and does not assume responsibility for them.

DATE OF PUBLISHING: 01 JUNE, 2020DATE OF POSTING

07-08 OF EVERY MONTH AT RMS POST OFFICE

Publishing Date || 01 June 2020 AGROBIOS NEWSLETTER

2 VOL. NO. XIX, ISSUE NO. 01

15. Organic Farming: A Review in Indian Context 26Ajit U. Masurkar and Akash D. Lewade

16. Silicon: An Element to Recover Stress Tolerance in Plants 28Mahesh Gurav and Rushikesh Pawar

17. Direct Seeded Rice: A Resource-Saving Technology forProfitableRiceProductioninIndia 29Manoj K. N.

18. Importance of Panchgavya in Crop Production 30Sunil and Seema Dahiya

19. Major Constrain in Pulses Production and Productivity in India 31Gaurav Shukla and Durgesh Kumar Maurya

20. Agronomic Weed Management: An Approach for Sustainable Crop Production 33Harsita Nayak

21. Physiological and Biological Aspects of Herbicide Activity and Selectivity 34Jagadish Jena, Twinkle Jena and Sameer Ranjan Misra

AGROMETEOROLOGY, REMOTE SENSING & GIS

22. Crop Cutting Experiments (CCE) 36G. Srinivasan, A. Karthikkumar and B. Sabarinathan

WEED SCIENCE

23. Cropping System: A Component in Cultural Method of Weed Management 38Varshini S. V.

ORGANIC FARMING

24. Potentialities of Use of Liquid Manures in Agriculture 39Avijit Ghosh, Manjanagouda S. Sannagoudar, Hanamant M Halli and Keerthi, M. C.

CROP ECOLOGY AND ENVIRONMENT

25. Carbon Sequestration 40Amrutlal R. Khaire, Prasantakumar Mazhi and Sonali V. Habde

CROP PHYSIOLOGY

26. Physiological Role of Chitosan in Plants 41Binny Sharma and Payal Chakraborty

27. Yellowing of Soybean and its Management from Farmer’s Perspective 43L. S. Rajput, Sanjeev K and V. Nataraj

28. Plant Phenomics: An Emerging Contemporary Approach to Combat Abiotic Stress in Crops 44Akankhya Guru, Soumya Kumar Sahoo, and Selukash Parida

29. Salinity: Key Hurdle in Crop Production 46Megha

CLIMATE CHANGE

30. Technologies for Carbon Sequestration in Agricultural Ecosystem 47Hemant Saini and Poonam Saini

31. Impact of Climate Change on Insect Population 48M. Thiyagarajan and J. Kousika

BIODIVERSITY

32. Importance of Traditional Knowledge in Maintaining Biodiversity 50Deepika Pandey

SOIL SCIENCE

33. Decomposition of Root Residues and Nutrient Release 50Biswabara Sahu, Arnab Kundu and Siddhartha Mukherjee

34. Soil Health Card: A Nuclear Mission Towards Sustainability in Agriculture 52Sudip Sengupta and Parijat Bhattacharya

35. Precision Nutrient Management 54Durgesh Kumar Maurya, Gaurav Shukla

36. Hyperaccumulators in Phytoremediation of Heavy Metals 56Pallavi. T. and Shwethakumari. U.

37. Bioavailability of Nutrients 57Poojitha K., Prashanth D. V., and Harsha B. R.

38. Humus and its Relavences on Soil, Plant and Environment 59Prashanth D. V., Harsha B. R. and Poojitha K.

39. Role of Sulphur in Indian Soils 60Ambika Prasad Mishra

40. Soil Sampling and Testing: A Tool for Soil Management 62Roohi and Hardeep Singh Sheoran

HORTICULTURE

41. Salicylic Acid: A Potential Phytochemical for Sustainable Fruit Production 63Farhana Khatoon, Dr Hidayatullah Mir and Khushboo Azam

42. Seed Production Technical Process in Special Reference Cucurbits 65Dr. Rakesh Kumar Meena and Tarun Nagar

43. Grafting in Solanaceous and Cucurbitaceous Vegetable Crops 66Umesh, B. C. and Jeevitha, D.

44. Hydroponics: The Future of Fruit Farming 68Hidayatullah Mir and Preeti Singh

45. Hydroponics: Advanced Technique of Vegetable Crop Production 69Khyati Singh and Mukesh Kumar Mehla



MEDICINAL AND AROMATIC PLANTS

46. Amazing Amla Juice 71Dr. Harpreet B. Sodhi

FORESTRY

47. Calliandra (Calliandra calothyrsus) Fodder: A Protein Supplement 72Chichaghare AR

PLANT BREEDING AND GEENTICS

48. Apomixis In Crop Improvement 73Tabinda Ali and Vartika Budhlakoti

49. RNA Interference: A Novel Approach for Crop Improvement 75Pawan Kumar Manoj Kumar and Sandhya Kulhari

50. Model Organisms for Genetic Studies 78Prasanta K. Majhi, Mounika Korada and Sonali V. Habde

51. Molecular Cross-Talk between Ethylene and Erfs for Submergence Tolerance in Rice 79Korada Mounika, Prasanta Kumar Majhi and Rashmi K

52. Crop Wild Relatives of Maize, Rice and Cotton for Tolerance to Abiotic Stress 80Basavaraj P S

53. Transgenic Male Sterility in Crop Plants: A Review 82Dr. G. B. Sawant, Dr. G. R. Gopal and Dr. S. G. More

54. AnInnovativeTechnique:ArtificialSeed 85Tamilzharasi, M.

55. Optical Mapping 87Soham Hazra and Shouvik Gorai

56. Brown Planthopper: A Serious Threat to Rice Production 88Ishwarya Lakshmi VG and Basavaraj PS

57. Lathyrus a Crop with its Nutritious and Toxic Effect 90Amrita Giri and Minu Mohan

58. Distant Hybridization in Crop Plants 91Versha

59. Role of Cell Signaling in Crop Improvement 92Meenakshi Rathi

SEED SCIENCE AND TECHNOLOGY

60. Homa Farming and its Uses in Agriculture 94Islavath Suresh Naik

61. Diagnosis of Seed Borne Pathogens 96Sunil Kumar

62. X-Rays in Seed Science and Technology 97C. Tamilarasan and L. Anilkumar

63. Genetic Use Restriction Technology 98Sridevi Ramamurthy

64. Halogen Dry Seed Treatment 100N. Vinothini, Poovarasan T.,

Bhavyasree R. K., and M. Sakila

PLANT PATHOLOGY

65. Fusarium Wilt (Tropical Race 4): A Destructive Disease of Banana in India 101Ajit Kumar Savani, K. Dinesh and Puli. Shashank Roy

66. Life Cycle of Pythium 102Ashutosh C. Patil and Ananta G. Mahale

67. Metabolome of Trichoderma: A Novel Strategy for Plant Defense and Growth 104Puli Sasanka Roy, Ajit Kumar. Savani and K. Dinesh

68. Plant Metabolite Engineering / Manipulation for Plant Defence 106Bandana Saikia, S. Ajit Kumar1 and K. Dinesh

69. Ontogenic Resistance 107P. Avinash

70. Post-Harvest Diseases Caused by Abiotic Factors 108P. Valarmathi

DISEASE MANAGEMENT

71. Management of Rice Diseases 109Prince Kumar Gupta, Sadhna Chauhan, Manoj Kumar Chitara and Sneha Shikha

NEMATOLOGY

72. Horizontal Gene Transfer is a Blessing for Nematode Parasitism Towards Plants 111Pranaya Pradhan and Jyoti Prakash Sahoo

ENTOMOLOGY

73. A Novel War Dance of the Honeybee 113R E Karthick, M. S. Sai Reddy, Somala Karthik and Manoj Kumar

74. Weather based Pest Forecasting 114Neeru Dumra

75. Ecological Threats for Pollinators and their Conservation 116Tara Yadav and Richa Banshiwal

76. Biology and Management of Cigarette Beetle, Lasioderma serricorne (F.) 117Jai Hind Sharma and Gaurava Kumar

77. Beeswax, a Miracle of Bee Hive: Its Production, Compositions and Uses 119Saswati Premkumari, Sabuj Ganguly and Rashmi Manohar Mahalle

78. Lesser Grain Borer, Rhyzopertha Dominica: A Major Threat to Storage Grains in India 120R. Tamilselvan, Banka Kanda Kishore Reddy, J. Kousika and S. Karthikeyan

79. EffectsofPesticidesonWildlife 122Banka Kanda Kishore Reddy, N. S. A. Devi, J. Kousika, S. Karthikeyan and R. Tamilselvan



80. Method Validation for Pesticide Residues Analysis 124S. Karthikeyan, Banka Kanda Kishore Reddy, J. Kousika and R. Tamilselvan

81. The Sterile Insect Technique 125Harpreet Sekhon and Ravinder Nath

82. The Story of Monolake and Alkali Fly 127Safeena Majeed, A. A.

INTEGRATED INSECT PEST MANAGEMENT

83. Endophytes in Plants Defence 128Bhagyasree S N, Suresh M Nebapure and Sagar D

POST-HARVESTMANAGEMENT

84. EffectofProcessingTechnologiesonFingerMillet 130Susrita Sahu and Rukeiya Begum

ECONOMICS

85. Land Holding in India: An Insight 131Shubhi Patel and Anju Yadav

86. Impact of Corona on Farm Produce 133Shwethakumari U and Kiran S C

COMPUTER ADDED TECHNOLOGY

87. Mobile Apps in Agriculture 133Dr. M. Kalpana and Dr. R. Parimalarangan

NATURAL RESOURCE MANAGEMENT

88. Bioremediation Techniques for use of Sewage Water 135Veeresh Hatti, Ashok K. Saini, Sanjay, M. T., Narayan Hebbal

EXTENSION EDUCATION AND RURAL DEVELOPMENT

89. Stubble Burning: Causes, Consequences and Alternatives 137Pawan Kumar Gautam

90. Agricultural Crisis and Lessons Amidst Corona Pandemic in India during Initial Lockdown 138Alok K Sahoo and Tarak C Panda

91. Role of Eco-Friendly Agricultural Practices used by Tribal Farmers in Crop Production for Sustainable Development 140Sonam Upadhyay

HOME SCIENCE

92. Sleeping Ergonomics: What Sleeping Position are You?? 142Divya Martolia

FOODS AND NUTRITION

93. Value Added Dairy Options 144Stephy Das and Dr. Manju K. P.



BIOTECHNOLOGY

20113

1. A New Pillar for Cancer Treatment: Reverse Immunology (Nobel Prize Winning Topic-2018)RAMACHANDRA ANANTAPUR* AND ROSHNI M

Ph.D. Department of Plant Biotechnology, UASB, GKVK, Bengaluru-65. *Corresponding Author Email: [email protected]

CancerA group of diseases caused by uncontrolled cell proliferation and migration is known as cancer.

More than 18 million people in the world are estimated to be diagnosed with cancer during the year 2018 (Global Cancer Observatory, 2018). Cancers are divided into different forms according to the cell type which include- Carcinomas, Lymphomas, Leukemias, Sarcomas and Brain tumors.

As mentioned above different types of cancer cause show different symptoms like rapid or slow rate of cell growth, tumors (visible growths) and some are asymtomatic. All the body cells follow the phenomenon of apoptosis, where cells old cells are killed and replaced by new cells, maintained by a regular signaling instructions by the body, cancerous cells lack this property. As a result, they continue to divide, multiply and affect growth of other normal cells.

TreatmentsInnovative research has fueled the development of new medications and treatment technologies. They are as

following- � Chemotherapy � Hormone therapy � Immunotherapy � Precision medicine � Radiation therapy � Stem cell transplant � Surgery � Targeted therapies

Checkpoint PathwaysCheckpoint pathways act as checks and balances that allow the immune cells to evaluate the attack. The pathways essentially function as the “brakes” when the body determines the response is no longer needed. The cancer cells will trick the immune system into turning on “checkpoint pathways”. The cancer can shut down the attack by utilizing these brakes and continue to grow. Blocking the effect of these checkpoint pathways can restore the normal function of the immune cells.

T cell activation and the concept of costimulationT cells have been at the center stage in immunology, but until the 1980s their antigen recognizing receptor remained unknown. Whereas the antigen-specific receptor on B cells was already well characterized. In the 1980s, T cell receptor (TCR) was identified and its interaction with MHC-associated peptides on antigen-presenting cells was revealed. (Linsley et al., 1992) suggested a role for costimulationin tumor immunology by demonstrating that the transfer of the B7 gene to tumor cells caused rejection. The first cell surface molecule identified in TCR costimulation was CD28. CD28 was found to synergize with the TCR complex in the activation of T cells (Martin et al., 1986). Four years later a ligand for CD28 was identified as the B7 molecule (now known as CD80), which is expressed on antigen-presenting cells.

Accelerators and brakes in our immune systemOur immune system functions in such a way that it discriminates between self and nonself entities in the body, to avoid and eliminate the externally invading bacteria, viruses and others harmful antigens. Among all the immune cells T cells play a vital role in in the defense, these T cells also need the accelerator proteins for complete defense action. T cells have receptors



which will bind to nonself type cells, hence trigger the immune system and engaging in defense mechanisms.

Many discoveries over this aspect contribute for the basic research of T cells, in identification of proteins that function as brakes, inhibiting immune activation. The balance between accelerators and brakes is very crucial for the control of immune system attack on invading antigens instead leading to autoimmune destruction of healthy cells and tissues.

A new principle for immune therapyDuring 1990s, James P. Allison studied the T-cell protein CTLA-4, in his laboratory at the University of California, Berkeley. He was one of several scientists who had made the observation that CTLA-4 functions as a brake on T cells with an entirely different idea on his mind, where he developed an antibody that could bind to CTLA-4 and block its function. He also observed the effect CTLA-4 blockade the T-cell brake which would stimulate immune system to attack cancer cells. Allison et al., performed a first experiment at the end of 1994.

By conducting the animal trails on mice, cancer was cured in antibody treated mice leading to inhibition of the brake and unlocking antitumor T cell activity. This research was advanced for human trails by the help of pharmaceutical industry. In 2010, a clinical study was conducted and results were astonishing, where the signs of cancer was disappeared in the patients with advanced melanoma.

The new form of immunotherapy i.e., inhibition of negative immune regulation unleashes a vigorous and often durable immune response directed against essentially any tumor already recognized by the immune system. This demonstrates how influential discoveries and findings have conferred great benefit on mankind and adds a new pillar to the existing cancer treatments.

Identification of CTLA-4 as a Negative Regulator

Incidentally the same year as the CD28 gene was isolated, the cDNA for a T cell expressed CD28-related molecule, CTLA-4. CTLA-4(Cytotoxic T Lymphocyte

Antigen. “CD152”CD28 and CTLA-4 belong to the immunoglobulin superfamily. It is now known that CTLA-4 protein resides intracellularly in resting T cells, but translocates rapidly to the membrane after activation (Lindsten et al., 1993; Linsley et al., 1996). In contrast to other T cells, it is constitutively expressed as a membrane protein in regulatory T cells (Takahashi et al., 2000).

� The hypothesis that blocking of CTLA-4 would strengthen the T cell anti-tumor response. Even pre-established tumors were found to be sensitive. Rejection was followed by durable tumor immunity. Two types of tumors responded for treatment

Discovery of the PD-1 receptor and its role in immune responses

� Honjo and colleagues assumed that PD-1, identified by subtractive hybridization to isolate mRNAs overexpressed in dying mouse cells, would be involved in pathways regulating apoptosis. Hence, they used the acronym PD (for Programmed Cell Death). ORF predicted protein with a transmembrane region, was distantly related to the immunoglobulin gene superfamily.

� PD-1 belongs to the extended CD28/CTLA-4 family of receptors, and that the cytoplasmic tail contains an essential “immunoreceptor tyrosine-based switch motif (ITSM)”. The ligand of PD-1 was discovered and named as PD-L1. Induce immune responses, also to tumors that did not express detectable PD-L1 or PD-L2. Even more



efficient as CTLA-4 therapy to treat tumors. Lead to less severe autoimmune side effects than CTLA-4 (Iwai et al, 2005).

PD-1 Blockade as a Treatment of Cancer

MEDAREX: An anti-CTLA-4 IgG1 monoclonal antibody named MDX-010 was developed in later named ipilimumab.

� The first marketing and manufacturing approval was granted 2014 in Japan. Pembrolizumab and Nivolumab, for the treatment of unresectable or metastatic melanoma and also extended for squamous non-small cell lung cancer in 2015.

� Even approved in Europe for treatment of melanoma.This new form of immunotherapy unleashes a

vigorous and often durable immune response directed against essentially any tumor. A crucial aspect in the future development is to improve understanding of these mechanisms, as well as the ones leading to adverse events. There is intensive research going on for identification of key mechanisms and biomarkers which will improve clinical efficacy and safety. These findings have conferred great benefit on mankind and add a new pillar to the existing cancer treatments.

20128

2. Micropropagation: A Technique for Rapid Propagation of Disease-Free PlantsMINAKSHI ONKAR BIRAJDAR

Department of Plant Tissue Culture, HiMedia Laboratories Pvt. Ltd. Mumbai, India *Corresponding Author Email: [email protected]

Micropropagation is clonal in vitro propagation of plant using a small portion of the plant tissues like shoot tip, anther, ovary, leaf, germinating seeds, etc. under the aseptic condition on a sterile nutrient medium. It is the asexual method of plant propagation that does not rely on the seed. The principle behind the micropropagation is the presence of totipotency in the plant cells, which provides an ability to regenerate single cell into a whole plant. Nowadays, it is the most acceptable and widely used method of plant propagation in economically important crop plants like banana, date palm, oil palm, pineapple, rubber tree, sugarcane, strawberry, bamboo, rose, carnation, gerbera, etc., due to its potential to regenerate true to type, disease, and pest-free plantlets.

Steps Involved in Micropropagation

Stage 0: Selection of mother plantsIt is an essential step for producing good quality plantlets. The mother plant is plant whose tissues like meristem/shoot tip, seeds, leaf, rhizome were used as explants in tissue culture. It should be an excellent high yielding variety and maintained at the good condition with no contamination of disease-causing agents.

Stage 1: Initiation of shoot culturesThe plant tissues excised from the selected mother

plants. A thoroughly washing with water and sterilizing agents like 70% ethanol, HgCl2 under an aseptic environment of laminar airflow. The tissues then transferred to MS media and kept in the culture room at 25± 2 °C in the dark for 3-4 weeks.

Stage 2: Multiplication of shoot-tip culturesThe regenerated callus was transferred to the multiplication medium, which is composed of MS media with different concentrations of IAA and BAP, and kept at 25oC temperature, relative humidity 50 - 80 % and 2-kilo lux intensity. The obtained shoots were sub-cultured for multiple rounds (usually seven to eight cycles) to produce multiple copies.

Stage 3: Root formationAfter getting an abundant number of shoot or shoot clumps, the single shoot should transfer to the rooting media for the promotion of root development. For rhizogenesis, auxin can be used from different sources like IAA, NAA or IBA with combination with MS media at between 0.1 and 2mg/l.

Stage 4: Planting out and HardeningHardening is one of the most crucial transition periods for the plantlets and carried out by altering external factors like water, temperature, and food sources as



nutrients supplied. Plants with roots and shoots are then first grown in the mist chamber having controlled conditions with 24-26°C temperature and more than 80 % humidity. Usually, initial hardening takes 2-3 weeks, then the plantlets were transferred in potting media and shifted to the greenhouse for 5-6 days. After this initial hardening, Press Mud Cake (PMC) mixed with soil is used for secondary hardening; it is an optimal medium which is supplemented with liquid NPK. After 2-3 months of hardening, the plants are ready for field plantation. All the stages of micropropagation depicted in figure 1.

FIGURE 1: Stages of micropropagation of plant tissuesA total of 9 months duration is required to get a

mature hardened plant ready for planting in the field from the inoculated explants. More than a thousand plants are generated from a single explant within a 9-month duration.

How Micropropagation is Boon for Plant Growers � High Rate of Plant Propagation from a small part

of the plant in a short period of time � Production of Disease-free Plants, virus-free

plants

� Cost-effective Process � Produce genetically identical plants. � Often produces healthier plants, leading to quicker

growth compared to those plants produced by a conventional method.

� Facilitates speedy international exchange of planting material, reduce quarantine period.

� Provides year around nursery for ornamental and fruit species.

� Secondary metabolite production which is a rich source of many pharmaceutical and industrial products.

� The backbone for Genetically engineered plant production with desirable qualitative and quantitative traits.

� Less workforce, small area, and less time required.

Applications of Micropropagation � Somaclonal variation � Germplasm conservation � Mutation breeding � Embryo culture � Haploid and dihaploid production � Somatic hybridization � Molecular farming � Genetic engineering � Disease-free plant production � Secondary metabolite production

Limitations of Micropropagation � Expensive requirement and sophisticated facilities

with a trained workforce � Chances of contamination � Vitrification. � Micropropagation is available only with limited

crop species

ReferencesChawla, H.S. 1998. Introduction to Plant Biotechnology.

Oxford & IBH Publishing Co. Pvt. Ltd., New Delhi.

20137

3. Recombineering: A Lambda Red Based Tool for Genetic Engineering*SURESH H. ANTRE AND SUNIL SUBRAMANYA A. E.*Ph. D. Scholar, Department of Plant Biotechnology, University of Agricultural Sciences, GKVK Campus, Bengaluru-560 065 *Corresponding Author Email: [email protected]

IntroductionRestriction enzyme cloning is the workhorse of molecular cloning, but its biggest limitation is the sequence modification can only be made at restriction site of the enzyme. An alternative method that can be used for cloning or genome engineering is the

lambda red system which is based on homologous recombination. It allows DNA to be directly modified within E. coli and it is independent of restriction sites. Recombineering is the short for homologous recombination mediated genetic engineering tool which uses lambda red system derived from the lambda red bacteriophage. It can be used to make a



variety of modifications such as point mutations or other small base pair changes, insertion and deletion of selectable and non-selectable sequence and the addition of protein tags. It also has the ability to modify the E. coli chromosome, plasmid DNA or BAC DNA.

To use the lambda red recombineering system to modify target DNA, first electroporate a linear donor DNA substrate (either dsDNA or ssDNA) into E. coli expressing the lambda red enzymes. These enzymes then catalyze the homologous recombination of the substrate with the target DNA sequence. Thus, cloning occurs in vivo, as compared to restriction enzyme cloning where the genetic changes occur in a test tube or in vitro. The donor DNA substrate takes only ~50 nucleotides of homology to recombine at the target site.

Key Components of the Lambda Red Recombineering SystemThe lambda red recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required for recombineering with a dsDNA substrate; however, only Beta is required when generating a modification with an ssDNA substrate.1. Gam: Gam prevents both the endogenous

RecBCD and SbcCD nucleases from digesting linear DNA introduced into the E. coli.

2. Exo: Exo is a 5’→3’ dsDNA dependent exonuclease. Exo will degrade linear dsDNA starting from the 5’ end and generate 2 possible products: 1) a partially dsDNA duplex with single stranded 3’ overhangs or 2) if the dsDNA was short enough, a ssDNA whose entire complementary strand was degraded.

3. Beta: Beta protects the ssDNA created by Exo and promotes its annealing to a complementary ssDNA target in the cell. Only Beta expression is required for recombineering with an ssDNA oligo substrate.

FIG.1 Components of Lambda Red Recombineering System (Fig. from Kenkel B, 2016)

Experimental OutlineA brief outline of a generic lambda red recombineering experiment is given below (Figure 2). In the following sections, key steps that differ from traditional restriction enzyme cloning will be explained in greater detail.1. Substrate DNA design and generation

2. Expression of lambda red recombination genes.3. Electroporation of substrate DNA and outgrowth

of bacteria.4. Selection and confirmation of recombinant clones.

FIG. 2 Overview of using Lambda Red Recombineering System to replace a gene of interest with an antibiotic resistance cassette (Fig. from Kenkel B, 2016)

Practical Uses of Lambda Red Recombineering System

� In vivo sub-cloning of DNA fragments from genomic DNA

� In–Out method of gene replacement � Gene replacement with selection marker

elimination by site-specific recombinase � Scar-less gene replacement or scar-less large-

scale deletions

ReferencesSharan SK, Thomason LC, Kuznetsov SG, Court DL.

Recombineering: A Homologous Recombination Based Method of Genetic Engineering. Nature protocols. 2009, 4(2):206-223.

Murphy KC. λ (Lambda) Recombination and Recombineering. EcoSal Plus, 2016, 7(1):1-70.



20142

4. Transcriptomics: What and What for?NUSRAT PERVEEN1* AND HIDAYATULLAH MIR2

1Horticulture-Fruits and Horticultural Technology, ICAR-Indian Agricultural Research Institute, New Delhi, India 2Department of Horticulture (Fruit and Fruit Technology), Bihar Agricultural University, Sabour *Corresponding Author Email: [email protected]

IntroductionThe Central Dogma of Molecular Biology explains the flow of genetic information within a biological system in a two-step process wherein first the information present in DNA, the permanent, heritable repository of genetic information, is transcribed into RNA, a transient intermediary molecule in the information network and then, a subset of RNA, the messenger RNAs are translated into protein. The complete set of DNAs, including all the coding and non-coding genes is termed as genome while the full complement of mRNA molecules produced by the genome under specific circumstances, at a specific developmental stage or in a specific cell is referred to as transcriptome, and it’s study is known as transcriptomics (Blumenberg, 2019). More often transcriptome is not the measure of absolute RNA levels, but rather comparison of relative levels within and/or between samples and can never be seen as a complete system in vivo since, all genes do not express simultaneously, in the same tissues, at the same levels. Further, the complexity of transcriptome increases by the processes like alternative splicing and RNA editing, making each gene capable of giving rise to many transcripts, each of which may have a unique expression profile.

MethodologiesTwo major strategies being used for transcriptome analysis include the direct sampling of sequences from source RNA populations or cDNA libraries, or from sequence databases derived there from i.e. RNA sequencing (RNA-seq), which uses high-throughput sequencing to capture all sequences (open system) and hybridization analysis with comprehensive, non-redundant collections of DNA sequences immobilized on a solid support i.e. microarrays, which quantify a set of predetermined sequences (closed system). Trinity, CLC genomics workbench (Qiagen—Venlo, Netherlands) and SOAPdenovo-trans are the major RNA-Seq de novo assembly software being used for whole transcriptome analysis.

Direct sequence sampling to quantify Steady-state mRNA levels: The first global gene-expression studies were based on the large-scale sequencing of Expressed Sequence Tags (ESTs) from cDNA libraries. An EST is a short nucleotide sequence generated from a single RNA transcript where RNA is first copied as cDNA by a reverse transcription and the resultant cDNA is sequenced using high-

throughput methods such as sequencing by synthesis (Solexa/Illumina, San Diego, CA). Further, Serial analysis of gene expression (SAGE) was developed as an improvement over EST to increase the throughput of the generated tags (Velculescu et al., 1995) which involves the generation of very short ESTs (9–14 nt), known as SAGE tags, which are joined into long concatemers that are cloned and sequenced using low-throughput, but long read length methods such as Sanger sequencing. These tags can be aligned to identify their corresponding gene if a reference genome is present otherwise can be directly used as diagnostic marker. The Cap analysis of gene expression (CAGE) method is a variant of SAGE that sequences tags only from the 5´ end of an mRNA transcript which can be useful for promoter analysis and for the cloning of full-length cDNAs.

DNA microarrays: DNA microarrays comprise a series of short nucleotide oligomers, known as “probes,” arrayed on a solid substrate which are hybridized (interrogated) with a complex fluorescently labelled probe (a probe comprising many different sequences, prepared from an RNA population from a particular cell type or tissue) and the fluorescence intensity acts as an indication of the transcript abundance. Prior genomic information in the form of annotated genome sequence or ESTs library is required for the development of the probes.

Massively parallel signature sequencing (MPSS): It is a hybrid between microarray technology and sequence sampling, in which millions of DNA tagged microbeads are aligned in a flow cell and analyzed by fluorescence-based sequencing.

Applications of TranscriptomicsTranscriptome profiling can be used to gain important insights and annotation of gene function by determining where and when particular genes are expressed. Differential expression of genes under stressed (viz. salinity, disease, drought) and unstressed conditions can facilitate the understanding of underlying mechanisms along with identification of key genes as a potential target for biotechnological manipulation with the aim of enhancing stress tolerance in plants and development of DNA markers linked with traits of interest. Transcriptome analysis also find an application in the bio-medical research including disease diagnosis and profiling. RNA-Seq approaches have led to identification of transcriptional start sites,



disease-associated single nucleotide polymorphisms (SNP) and novel splicing alterations having a bearing on human diseases thus, facilitating the interpretation of disease-association studies. Progression of cancer has been attributed to various abnormal splicing events and hence, monitoring changes in gene expression can be useful for identifying cancer-causing genes or regions of transcriptome that are responsive to treatment.

ConclusionIn the past few decades, the understanding of genome expression has been revolutionized by transcriptomics. The development of high throughput, low cost sequencing techniques and increased availability of genome sequence information has made it possible to compare transcriptomes of thousands of organisms, tissues, or environmental conditions thus providing a comprehensive insight into the complexities of cellular world. The large amount of data generated

by transcriptomics studies, in the raw or processed form are submitted to public databases to ensure their utility for the broader scientific community. Some of the public transcriptome databases include Gene Expression Omnibus, Expression Atlas and Genevestigator containing data in the form of Microarray, RNA-Seq maintained by the hosts, National Center for Biotechnology Information (NCBI), European Bioinformatics Institute (EBI) and privately curated respectively.

ReferenceBlumenberg, M., 2019. Introductory Chapter:

Transcriptome Analysis, Miroslav Blumenberg, IntechOpen, Available from: https://www.intechopen.com/books/transcriptome-analysis/introductory-chapter-transcriptome-analysis

Velculescu, V.E., Zhang, L., Vogelstein, B and Kinzler, K.W. 1995. Serial analysis of gene expression. Science. 270:484–487.

20156

5. Combined Approach of Morphological and Molecular Diagnosis of Diseases in Plants*BANSULI

Ph.D. Scholar, Department of Agricultural Biotechnology, CSK HPKV, Pamapur, Himachal Pradesh, India, 176062 Corresponding Author Email: [email protected]

IntroductionFood security has become an important international issue in recent years. The demand for food will continue to increase due to rapid increase in human population. One of the most prominent reasons behind the food crisis is decrease in agricultural productivity. Spoil caused by pathogens and pests in plants plays a considerable role in crop losses. In order to minimize the damage caused by pathogens and pests in crops during growth, harvest, postharvest stages and to increase the productivity rapid and advanced disease detection methods are highly significant.

The morphological diagnosis of disease in plants quickly identifies limited number of species associated with particular host or disease. Identification and detection of diseases in crops can be done with conventional and molecular methods. Conventional methods include histopathological (on the basis of infected tissues) mostly done in cases of fungi, bacteria and nematodes, Culture growth/colony characters, microscopy, staining and differentiation on basis of bio-chemical properties. In current years, molecular techniques of plant disease detection have been well recognized and are more reliable. The molecular techniques are very sensitive as requires minimum amount of microorganism that can be detected. The frequently used molecular techniques for disease

detection are ELISA and PCR. Other molecular techniques include immunoflourescence (IF), flow cytometry, fluorescence in situ hydridization (FISH) and DNA microarrays.

Conventional MethodsConventional methods are done by observing the symptoms appeared on the host tissue either by naked eye and hand lens. All plant pathogens produce distinctiveness symptoms in different parts of the plants and that can be can be detected through various conventional methods as listed below.

� Histopathological methods � Culture growth/ colony characters � Microscopy � Differentiation on basis of bio-chemical properties

Advance Methods of Disease DetectionThere are two types of advanced methods of disease detection in plants.

� Direct methods � Indirect methods

Direct methods include laboratory-based techniques such as polymerase chain reaction (PCR), immune fluorescence (IF), fluorescence in-situ hybridization (FISH), enzyme-linked immune sorbent assay (ELISA), flow cytometry (FCM) and



gas chromatography-mass spectrometry (GC-MS), whereas thermography, fluorescence imaging and hyperspectral techniques are used under indirect methods.

Biosensors based on enzyme, antibody, DNA/RNA and bacteriophage as a new tool for the early identification of crop diseases. The flow chart of disease detection methods is showed in figure 1.

FIGURE 1. Different disease detection methods

Serological methodsSerological methods are used for high-throughput analysis for large numbers of samples. In these methods the disease-causing pathogens such as bacteria, fungi and viruses are directly detected to provide accurate identification of the disease/pathogen. Some

serological methods are listed below. � Flow Cytometry � Enzyme-Linked Immunosorbent Assay (ELISA) � Immunofluorescences (IF) � Fluorescence in-situ hybridization (FISH)

Molecular methodsDirect detection of diseases includes various molecular methods that could be used for high-throughput analysis. Based on DNA hybridization and replication, PCR was initially used for highly specific detection of diseases. Now it has been extensively used for the detection of plant pathogens. In addition to the basic PCR technology, advanced PCR methods such as reverse-transcription PCR (RT-PCR), Multiplex PCR and Real-time PCR has also been used for plant pathogen identification due to its high sensitivity. Some broadly used molecular methods are given below.

� Polymerase Chain Reaction � Multiplex RT-PCR � LAMP technique � Real-time RT-PCR � DNA-Arrays

Indirect methods � Thermography � Fluorescence Imaging � Hyperspectral Techniques � Gas Chromatography

Detection of plant diseases using portable sensors

� Bacteriophage-Based Biosensors � Affinity biosensors � Antibody-Based Biosensors � DNA/RNA-Based Affinity Biosensor

20174

6. Abiotic and Biotic Stress Signal Cross Talking in PlantsJYOTI PRAKASH SAHOO1*AND UPASANA MOHAPATRA2

Ph.D. Research Scholar, 1Department of Agricultural Biotechnology, OUAT, Bhubaneswar 2Department of Plant Biotechnology, UAS, GKVK, Bengaluru *Corresponding Author Email: [email protected]

Cross Talk BiologyBiological crosstalk alludes to cases in which at least one or more components of one signal transduction pathway influences another. This can be accomplished through various ways with the most widely recognized structure being crosstalk between proteins of signaling cascades. In these signal transduction pathways, there are regularly shared segments that can communicate with either pathway. A progressively perplexing example of crosstalk can be seen with transmembrane

crosstalk between the extracellular matrix (ECM) and the cytoskeleton. A case of crosstalk between signalling pathways is component of cAMP/PKA restraint of ERK initiation (MAPK pathway) where cAMP actuation of PKA enacts Rap1 by means of Src and Rap1, then phosphorylates Ras and hinders motioning of signals to Raf-1.

Another case of crosstalk between signalling pathways is lymphocyte enactment. Indeed, even without enactment by a ligand bound to the receptor



(R1), the MAPK pathway typically shows basal action (at low levels). In any case, HePTP balances this movement. It prompts initiation of the cAMP pathway by binding of ligand to its appropriate receptor (R2) prompts the actuation of cAMP-subordinate protein kinase (PKA) by adenylate cyclase (AC) which enacts PKA then phosphorylates HePTP at Ser23, repressing its capacity to tie to Erk and subsequently hinder the MAPK pathway. A chat of plant in which activation of CDPK and MAPK Cascades by various stresses are presented in Table 1.

Drought-Stress or Cold-Stress Signal Transduction Pathways in Arabidopsis thalianaThere are six signal transduction pathways: two are ABA-reliant and four are ABA-independent. Stress inducible genes rd29A/cor78/lti78, rd29B/lti65, rd22, and erd1 have been utilized to break down the guideline of quality articulation and the signalling procedure. abi1, abi2, and era1 are engaged with ABA signalling. hos5 works in DREB2-related drying out (dehydration) signalling, and sfr6, hos1, and hos2 work in DREB1/CBF-related cold signalling. esk1 is associated with reactions to cold by means of a DRE-free procedure. Two cis-acting components, DRE/CRT and ABRE, are associated with the ABA-free and ABA-responsive acceptance (induction) of rd29A, individually. Two diverse DRE/CRT-restricting proteins, DREB1/CBF1 and DREB2, recognize two distinctive signal transduction pathways because of cold and drought stresses, separately. DRE/CRT-restricting proteins contain an AP2 DNA-binding domain, while ABRE-restricting proteins encode bZIP translation factors.

Transcriptional Regulation of Stress-Responsive Genes during Osmotic Stress in PlantsOsmotic pressure (stress) instigated DREB2 interpretation factors incite ABA-independent translation of stress responsive genes. ABA-dependent pathways direct stress responsive genes articulation through CBF4, MYC/MYB and bZIP-type transcription interpretation factors, which tie to the CRT/DRE, MYB/C Recognition Sequences (MYB/CRS) and ABA Responsive Elements (ABRE) advertiser components, separately. ABA-dependent abiotic stress signalling is interceded partially through IP3 and Ca2+. FRY1 contrarily directs IP3 levels. ABA-instigated (induced) Ca2+ signalling is negatively regulated by the ABI1/2 protein phosphatase 2Cs and the SCaBP5±PKS3 complex

Regulation of a Cold-Stress-Responsive Transcriptome and Freezing Tolerance in PlantsICE1 is a constitutively communicated myc-like bHLH interpretation factor, which is dormant under non-stress conditions. Low temperature stress apparently actuates ICE1, which ties to myc cis-elements of the CBF3 promotes CBF3 expression. CBFs ties to the CRT/DRE cis-components on cold-stress-responsive (COR) genes inducing their expression and leading

to acquired freezing tolerance. HOS1, a putative E3 ubiquitin-conjugating enzyme, seems to focus on a positive controller of CBFs for degradation.

Ion Homeostasis for Salinity Tolerance in PlantsSalt stress actuated Ca2+ signals are perceived by SOS3 which initiates the SOS2 kinase. The SOS3±SOS2 kinase complex controls cell Na+ levels by invigorating Na+ transport out of the cytoplasm (for example by expanding the articulation and action of SOS1) and possibly by confining Na+ entry into the cytosol (for example restraining HKT1 movement). An extra objective of the SOS2 kinase, NHX (vacuolar Na+/H+ exchanger), additionally adds to Na+ particle homeostasis by shipping Na+ from the cytoplasm into the vacuole.

Reactive Oxygen Intermediate (ROI) Scavenging in Plants during Oxidative StressSuperoxide dismutase (SOD) acts as the first line of defense converting O2 into H2O2. Ascorbate peroxidases (APX), GPX (Glutathion Peroxidase) and CAT (Catalase) then detoxify H2O2. In contrast to CAT, APX and GPX require an ascorbate (AsA) and/or a glutathione (GSH) regenerating cycle which uses electrons directly from the photosynthetic apparatus or NADPH as reducing power.

Signal Cross Talking during Pathogen Defense in Arabidopsis thalianaSalicylic acid (SA), Jasmonic acid (JA) and Ethylene (ET) take part in the signal crosstalk during pathogen defense in case of Arabidopsis thaliana. The SID2 and EDS5 genes appear to be directly involved in SA biosynthesis whereas the EDS1, EDS4 and PAD4 genes regulate SA synthesis. The placement of EDS5 downstream of EDS1, EDS4 and PAD4 is based on studies indicating that the expression of EDS5 is dependent on these genes. The placement of COI1 and MPK4 early in the JA signalling pathway is based on reports that mutations in these genes block JA signaling. However, the phenotypes of these mutants are not identical; although both mutants exhibit defects in pollen development, mpk4 mutants constitutively express SA dependent genes, whereas coi1 mutants do not. The placement of COI1 and MPK4 with respect to each other in the JA signalling pathway awaits further molecular and genetic experiments.

TABLE 1: Activation of CDPK and MAPK Cascades by various stresses in plants

Plant Name of CDPKs CDPK Expression

Arabidopsis AtCDPK1, AtCDPK2

Induced by drought and salinity but not induced by cold or high temperature (Urao et al., 1994)

Arabidopsis AtCDPK1 ABA Inducible (Sheen,1996)




Rice OsCDPK7 Induced by cold and salt stress (Patharkar et al., 2000)

Plant Name of MAPKs MAPK ExpressionAlfaalfa SIMK Salt Induced (Munnik et

al., 1999)Tobacco SIPK Salicylic acid induced

(Hoyos et al., 2000)Arabidopsis ATMPK3,

ATMPK3 and ATMPK3

Salt stress (Mizoguchi et al., 2000)


Rice OsMAPK5 ABA, Biotic and Abiotic Stress (Xiong, 2003)

ReferenceKnight, H., & Knight, M. R. (2001). Abiotic stress

signalling pathways: specificity and cross-talk. Trends in plant science, 6(6), 262-267.

Mittler, R. (2002). Oxidative stress, antioxidants and stress tolerance. Trends in plant science, 7(9), 405-410.

20194

7. Cisgenesis: Can it be an Alternative to Transgenesis?UPASANA MOHAPATRA1* AND JYOTI PRAKASH SAHOO2

Ph.D. Research Scholar, 1Department of Plant Biotechnology, UAS, GKVK, Bengaluru 2Department of Agricultural Biotechnology, OUAT, Bhubaneswar *Corresponding Author Email: [email protected]

IntroductionThe increasing demand of good quality and large quantity food impel the researcher to discover new alternative strategies for crop improvement. Though the application of molecular biology as a novel approach for crop improvement is being done since three decades but the genetic modification of crop through transgenesis is still not acceptable due to ethical, social and environmental safety issues. So these constraints in transgenesis necessitate to uncover new approach for crop improvement which is ‘Cisgenesis’.

CisgenesisCisgenesis was first introduced by Jochemsen and Schouten in 2000 in their published book titled as” Toetsen en begrenzen. Een ethische en politieke beoordeling van de moderne biotech-nologie. In this book they proposed the principles of cisgenesis, that the genes or gene elements should be derived from the species that was to be genetically modified itself. Additional sequences, including introns or the regulatory sequences originating from the same gene as the coding sequences, were not necessary.

In 2006, Schouten et al., modified the definition and proposed cisgenesis as the integration of one or more genes (along with introns, flanking regions such as native promoter and terminator regions in a sense orientation) isolated from a crossable donor plant. Along with cisgenesis intragenesis concept coexist though intragenesis differs from cisgenesis in the aspect of gene arrangement as the encoding sequence and regulatory sequence are derived from a different

gene in intragenesis. Though cisgenesis and classical plant breeding involve the gene transfer within the same or between evolutionarily close species but cisgenesis has a lot of advantages compared to classical breeding which are discussed as below.

Cisgenesis over conventional plant breedingThe classical breeding takes much longer time for transfer the gene of interest from doner plant to recipient plant whereas cisgenesis is faster and precise to transfer gene between the related species. In classical breeding the transfer of one or more undesirable gene along with the gene of interest makes the recipient plant useless. This can be avoided in the cisgenesis. In case of tightly linked gene linkage drag is impossible to remove the deleterious gene in the back crossed line. In such cases cisgenesis can be used as a clean introgression breeding only with the target gene. In conventional breeding the genetic makeup of the progeny is changed and varied from the parents while in cisgenic plant the genetic makeup is of the original recipient plant is maintained. The development of multigenic plant is quite tedious and time taking in conventional plant breeding whereas in cisgenesis more than one gene can be transferred in one insertion to the target plant. Higher level of trait expression is achieved in the cisgenesis by inserting more than one copy of the cisgene in the recipient plant

Cisgenic plants are different from the transgenic plantThough each and every technique of transgenesis is



used in cisgenesis the source of gene brings out the difference between transgenic and cisgenic plant. In the transgenesis the gene is derived from unfamiliar, sexually incompatible species without considering the species barrier across the kingdom but cisgenesis respects the species barrier as the gene along with the regulatory element is inserted from the cross compatible species or its close relatives.

Integration of transgene may lead to the formation of a novel gene and in turn a new trait is appeared in the recipient plant which may not be achieved by conventional breeding. So transgenesis helps in extension of gene pool whereas the cisgenesis cannot. Transgenic plant may create the problem of unintentional transfer of novel genes to its cross compatible species which may affect the ecological balance. But in case of cisgenesis there is no transfer of genes to the nontargeted species and the cisgenic plants are ecologically safe. Also, in cisgenesis change in plant vigour is not observed as in transgenesis.

Cisgenesis in crop improvementThe first cisgenic plant was reported in apple for scab resistance by inserting the resistance gene HcrVf2 into Gala cultivar of apple. In New Zealand based company Cisgenic is used as a registered trademark for engineering the fodder crops like ryegrass and clover. The main goal of plant breeders to produce the crops for biotic and abiotic stress tolerance with the improvement of quality and quantity of food as well. A list of crops engineered through cisgeneis is mentioned in Table 1.

Limitations of CisgenesisThe trait exterior to sexually compatible species cannot be introduced. It requires efficient worker with extraordinary skill and more time. For development of marker free plants innovative protocols are required. Also, so much hard work is required to remove the cisgenic lines integrated with the sequences of vector backbone. The crops with low transformation and regeneration efficiencies are restricted from cisgenesis.

TABLE 1: List of some crops engineered through cisgenesis

Crop Donor species Trait GeneApple Malus floribunda Apple Scab

ResistanceHcrVf2

Barley - Phytase activity

HvPAPhy_a

Crop Donor species Trait GeneRye grass Lolium perenne Drought

toleranceLpvp1

Potato Solanum bulbocastanum

Late blight resistance

Rpi-blb1,Rpi-blb2,Rpi-blb3

Potato Solanum papita Late blight resistance

Rpi-pra1

Potato Solanum stoloferum


Rpi-sto1

Potato Solanum berthaultii


Rpi-ber1

Potato Solanum venturi Late blight resistance

Rpi-vnt1

Strawberry - Fruit rot (Botrytis cinerea)

PGIP

Poplar Populus trichocarpa clone Nisqually-1

Gibberellin metabolism

PtGA20ox7, PtGA2ox2, PtRGL1_1, PtRGL1_2 and PtGAI1

(Source: Telem et al., 2013)

ConclusionBased on the available research knowledge it can be assumed that cisgenic plants are different from transgenic plants and more or less similar to conventional bred plants. As cisgenic plants are free from toxic, undesired gene or any deleterious genes, the plants should be free from the regulation applied to the transgenic plants. By expanding the research area in cisgenesis and exempting it from regulation framework crop improvement through cisgenesis can able to drive the plant breeding in faster rate.

ReferencesDudziak, K., Sozoniuk, M., Kowalczyk, K., & Nowak,

M. (2019). Cisgenesis as a novel prospect for crop improvement. A review. Annales UMCS sectio E Agricultura, 74, 2.

Espinoza, C., Schlechter, R., Herrera, D., Torres, E., Serrano, A., Medina, C., & Arce-Johnson, P. (2013). Cisgenesis and intragenesis: new tools for improving crops. Biological Research, 46(4), 323-331.

Telem, R., Wani, H., Singh, B., Nandini, N., Sadhukhan, R., Bhattacharya, R. & Mandal, N. (2013). Cisgenics-a sustainable approach for crop improvement. Current genomics, 14(7), 468-476.



20211

8. IdentificationandDifferentiationofCrops through DNA Barcoding1*S. G. MAGAR AND 2V. G. MAGAR1*Ph.D. Research Scholar, Biotechnology Centre, Department of Agricultural Botany, Post Graduate Institute, Dr. P.D.K.V. Akola, (M.S.) 2*Assistant Professor, CSMSS, College of Agriculture, Kanchanwadi Aurangabad, VNMKV, Parbhani (M.S.) *Corresponding Author Email: [email protected]

IntroductionNowadays the basis of classification of taxa has progressively moved from organismal to cellular to molecular level. Molecular structures and sequences are most commonly used to revealing evolutionary relationships, expression studies, etc. Much of the plant identification is failing in species diagnoses, taxonomic expertise. There was need for Restructuring Systematics, the biological branch that deals with classification and nomenclature. Here come the DNA sequences as taxon ‘barcodes’. Plant DNA Barcoding permitting biological identification of plant species through DNA sequences. DNA barcoding is a technique used to identify unknown materials of known species based on DNA sequences of standard genome regions (Lahaye et al., 2008). Ideally barcodes are small DNA sequences highly variable, distinctive, easy experiment conduct and low cost (Dong et al., 2014). This method aims to provide species-level identifications, as such will contribute powerfully to taxonomic and biodiversity research. DNA barcodes can help expand

our knowledge by exploring many more species rapidly and inexpensively. The results obtained from DNA barcoding studies can then help us identify species that are good targets for more detailed genetic analyses.

� Barcoding Regions for Different Species Must Be Different:a) Ideally selection of single DNA locus which is

different in each species.b) Universality: to amplify selected DNA region

need primers that will amplify consistently.c) Robustness: need to select a locus that

amplifies reliably, and sequences well. � Selection of Source Tissue (Fresh, herbarium,

Xylarium): For plants generally plastid DNA are used for barcoding because DNA are short, easily alienable, maternal inheritance, easily recoverable from herbarium sample. Developing DNA barcoding reference libraries which tissue should be use its Pictorial representation given bellow with their potential strengths and weaknesses of source tissue

PIC 1. Selection of source tissue (source: Lichao Jiao et al. 2018)

� Standard DNA barcoding for plants:



PIC 2. Barcoding Protocol for Plants (source: internet)

Application1. Protection of Endangered Species2. Tracking adulterations3. Verification of plant products4. Identification of invasive species5. Works for all stages of life6. Identifying Agricultural pest7. Accurate species identification8. Characterization of below-ground plant diversity

using roots9. Used to create phylogenetic trees

ConclusionsDNA barcoding has emerged and established itself as a potential tool for species phylogenetics studies and identification. This powerful tool use in the context of quality control of both well and poorly regulated supply systems. It is used for authenticating products based on single herbal ingredients and DNA sequence

to assists species diversity in processed products. It’s been proving very useful to protect endangered species, tracking adulteration in products and sustaining environment.

ReferenceDong, W., Xu, C., Li, C., Sun, J., Zuo, Y., Shi, S., Cheng,

T., Guo, J. and Zhou, S. (2015). Ycf1, the most promising plastid DNA barcode for land plants. Sci. Rep., pp. 1-5.

Lahaye, R., Bank, M.V.D., Bogarin, D., Warner, J., Pupulin, F., Gigot, G., Maurin, O., Duthoit, S., Barraclough, T. G. and Savolainen, V. (2008). DNA barcoding the floras of biodiversity hotspots. National Academy of Sciences of the USA., 105(8): 2923-2928.

Lichao, j., Min, Y., Alex, C., Wiedenhoeft, Tue. H., Yafang, Y., (2018). DNA Barcode Authentication and Library Development for the Wood of Six Commercial Pterocarpus Species: The Critical Role of Xylarium Specimens. Science reports., 8: 1945.

MICROBIOLOGY

20114

9. Plastic Lovers: A Green thought for PlasticsMS. ASWATHY, J. C.1 AND DR. SHALINI PILLAI, P.2

1PG Scholar, Department of Agronomy, College of Agriculture, Vellayani 2Professor, Department of Agronomy, College of Agriculture, Vellayani

“We made plastic, we depend on it, now we are drowning in it”. These are the famous words of Laura Parker, writer for The National Geographic magazine. As she has said plastic is considered as the most useful invention man has made till date. But the excessive and uncontrolled use of it has turned it into a bane. Now we are at a point where it seems that we are living in a planet full of plastic. Scientists in the course

of time have come up with a lot of viable options to reduce the increasing amount of plastic pollution. One such option is the plastic lovers. Plastic lovers are those organisms that have greater affinity towards plastics and are capable of disintegrating it. Many organisms have been identified across the world. These include bacterium (Ideonella sakiensis), fungus (Pestalotiopsis microspora) and wax worm (Galleria mellonella)



Ideonella SakiensisIt is a gram negative, aerobic, rod shaped, non-spore forming bacterium. The plastic degrading ability of this bacterium was reported by a team of research workers from Japan in 2016. They collected about 250 samples contaminates with PET debris from different sites. The samples included soil, sludge and waste water from recycling plants. In this study they discovered a bacterial strain viz., Ideonella sakaiensis 201-F6 from one of the samples. It was identified that this bacterium was solely responsible for the degradation of the PET film at the rate of 0.13 mg cm–2 day–1. The entire microbial community was shown to mineralize 75 per cent of the degraded PET into carbon dioxide once it had been initially degraded and assimilated by I. sakaiensis.

The bacterium grows on PET surfaces by adhering with thin appendages. These appendages secrete PET-degrading enzymes onto the PET surface which results in the biodegradation of PET. The degradation was observed to be mediated by two enzymes - PETase and MHET hydrolase. The former enzyme is secreted outside the cell to break up the PET polymer into MHET (mono-(2-hydroxyethyl) terephthalate). This MHET is taken up by the bacterium and the latter enzyme hydrolyses the MHET, breaking it down into ethylene glycol and terephthalic acid (monomer units of PET). Ethylene glycol is readily taken up and used by I. sakaiensis and many other bacteria whereas the terephthalic acid, integrated into various metabolic pathways. Thus both the molecules are used by the cell to produce energy and to build necessary bio-molecules. However, this whole process was extremely slow in nature and took about 6 weeks for the complete degradation of PET into its monomers. Since the organism is aerobic in nature it is not suitable for cleaning up of the landfills. It’s not a complete solution to plastic pollution – but it does show how bacteria could help create more environmentally friendly recycling.

PLATE1. Ideonella sakiensis, copyright: http://sarkardaily.com

Pestalotiopsis microsporaThe designer Katharina Unger along with the

scientists from Utrecht University, identified the ability to degrade polyurethane in a rare fungus called Pestalotiopsis microspora. The discovery has been made from the Amazonian rainforests of Ecuador. This fungus was able to completely degrade polyurethane into organic matter. With the discovery of Pestalotiopsis microspora a striking combination of creativity, science and design was brought up and a viable option to clean up the landfills was identified.

PLATE 2. Pestalotiopsis microspora

PLATE 3. Spores of fungus Pestalotiopsis microsporaCopyright: http://funguys.co.za

Wax WormWax worm is the caterpillar larvae of wax moth (Galleria mellonella) and are medium-white coloured with black-tipped feet and small, black or brown heads. In the wild, it lives as nest parasites in bee colonies. The larvae feeds on the comb made by the bees causing the destruction of the comb whereby contaminating the stored honey, kill bee larvae and /or cause the spread of honey bee diseases.

The ability of wax worm to digest plastic was identified by Bertocchini along with with fellow scientists, Paolo Bombelli and Christopher. This was an accidental discovery made by Bertocchini when she was cleaning up the beehives kept at home. She collected the worms which were in the hives and tied it up in a shopping bag for discarding. But after an hour when she inspected she could find small holes all through the bag and the worms were out and there the study of another milestone discovery for the biodegradation of plastic began. Both wax and the polyethylene in Bertocchini’s plastic bag had a similar carbon backbone and the scientists reported that since



the larvae is capable of digesting wax they might have evolved enzymes to degrade plastic also which was having a similar structure.

PLATE 4. Caterpillar larvae of wax moth.Copyright: http://npr.org

PLATE 5. Wax worm feeding on polyethylene sheet.Copyright: http://theatlantic.com

When about 100 wax worms were placed in polyethylene shopping bags, they consumed almost 0.1 g of the plastic over the course of 12 hours under laboratory conditions (Bombelli et al., 2017). The wax worms metabolized the polyethylene plastic films into ethylene glycol, a compound which biodegrades rapidly under the normal environmental conditions. Since wax worm is a parasitic pest of beehives it cannot be used as such for the degradation of plastic and at present the scientists are working to isolate that particular enzyme responsible for the degradation of plastic from the worms.

ConclusionThough all these organisms have proved their ability to degrade plastic and thereby reduce the burden of plastic on the planet, at present none of these organisms are being used for the cleaning up of plastic. Since these are microorganisms studies are still being undertaken to identify whether they pose any harmful effects when released into the environment. Research is still in the pioneer stage to identify and isolate the enzymes present in these organisms, responsible for plastic degradation, so that they could be effectively used to curb the raising issue of plastic pollution.

BIOCHEMISTRY

20164

10. First and Second Genome of DNA SequencingPAYAL CHAKRABORTY AND BINNY SHARMA

Research Scholar, Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi *Corresponding Author Email: [email protected]

Genome SequencingGenome sequencing is searching out the exact order of DNA nucleotides sequence or bases like order of A, T, G, C in a genome of any living organisms.

TABLE: 1. History of genome Sequencing

Year Sequencing events1953 Discovery of DNA double helical structure by

Watson and Crick1970 Discovery of type II restriction endonuclease by

Hamitlon Smith1977 Discovery of Sanger method of genome sequencing1977 Discovery of Maxam-Gilbert method of genome

sequencing

Year Sequencing events1983 Discovery of Polymerase Chain reaction by Kary

Mullis1990 Cycle Sequencing, Improved Sequencing Enzymes,

Improved fluorescent detection scheme2002 NGS: 454 Pyrosequencing

First Generation SequencingThe two popular conventional method of genome sequencing include:1. Sanger or Chain Termination Sequencing method2. Maxam and Gilbert or Chemical Sequencing

method

Sanger Sequencing MethodThis method was developed by Frederick Sanger and



colleagues in 1977. After 25 years of its discovery he got NOBEL PRIZE in 1980. It requires single Stranded template, Primer, DNA polymerase, Deoxynucleotides (DNTPs), Di-Deoxynucleotides- (The 3′-OH group necessary for formation of the phosphodiester bond is missing in ddNTPs).

Procedure1. Denaturation of double stranded DNA2. Primer annealing and extension of bases with the

help of DNA polymerase and DNTPs.3. Chain Termination: Without the 3’ OH, no more

nucleotides can be added and DNA polymerization will cease.

4. Gel electrophoresis and Autoradiography are performed for Sequence Determination.

Advantages � Give unbiased result for small number of

sequencing samples. � Cost effective in case of small-scale sequencing.

Disadvantages � It can sequence short sequence of DNA about

300-1000bp. � Quality of sequencing degrades beyond 800bp.

Maxam-Gilbert Method of SequencingThis method was developed by Allan Maxam and Walter Gilbert in 1976–1977. This method is based on nucleotide specific chemical modification of DNA and subsequent cleavage of the DNA.

Procedure1. Denaturation of double stranded DNA by high

temperature and 5’ end is radioactively labeled by using ϒ-32P.

2. Cleavage of DNA strand by using chemicals like piperdine followed by Dimethyl sulphate cleaved at purine (A and G) and hydrazine which selectively cleaved pyrimidines (C and T). After chemical

treatment the cleaved Fragments contain four different nucleotide sets G, A+G, C and C+T. A+G. (Hydrazine, Hydrazine NaCl, Dimethyl Sulphate, piperidine cleaved at T+C, C, A+G, G respectively.

3. Four different nucleotide sets are placed in four different test tubes and gel electrophoresis is carred out in acrylamide gel that separate molecules according to their shape and size.

4. A series of dark band is obtained that locate the position of DNA molecules when the gel is placed under X-ray.

5. After ordering the DNA fragments by size a sequence of DNA molecule is obtained.

Advantages � It is used in DNA-protein interactions like Foot

printing. � It used in determining nucleic acid structure and

epigenetic modifications

Disadvantages � This technique cannot be used in sequencing long

base pairs of DNA � Use of toxic chemicals and complex process.

TABLE: 2. Comparisons between Sanger and Maxam Gilbert Sequencing:

Sanger Method Maxam GilbertIt is developed by Frederick Sanger in 1977

It is developed by Allan Maxam & Walter Gilbert in 1977-80

It is a enzymatic method It is chemical methodDNA synthesis is required DNA is requiredIt include termination of chain elongation

DNA is cleaved at different nucleotides positions

Automation is available Automation is not availableLess specific and sensitive

More specific and sensitive

Requirement of less toxic chemicals

Require toxic chemicals

NANOTECHNOLOGY

20103

11. Nanobiosensors: An Emerging Technology in AgricultureMOHAMMED NISAB C. P.

Department of Soil Science and Agricultural Chemistry, Palli Siksha Bhavana, Visva-Bharati.

Accessibility of net land and water-resources for agriculture is rapidly declining, causing huge loss in agricultural output. These issues can only be dealt efficiently with the aid and continuous flow of new technologies. Presently, nanotechnology is visualized

as a rapidly evolving field with high potential to revolutionize agricultural system. Nanotechnology is the design, fabrication and use of nano structured systems and the growing, assembling of such systems mechanically, chemically or biologically to form nano



scale systems and devices.Biosensor is a sensor that integrates a biological

element with a physiochemical transducer to produce an electronic signal proportional to a single analyte which is then conveyed to a detector. A nanobiosensor is usually built on the nano-scale to obtain process and analyze the data at the level of atomic scale. It is an analysis tool is described as dense bio-sensors, bio or mimic the biological sensing element with a company that is close to, or in contact with the transducer mounted on it. Diagnosis is based on the analysis of specific contacts with biological recognition element is based. In the year 1967, the first biosensor was invented which led to the development of several modified biosensors. Interestingly, since early 20th century the concept of biosensors existed but their uses were limited only in laboratories and with advent of sciences several modern biosensors were designed.

Overall, there are three so-called ‘generations’ of biosensors; first generation biosensors operate on electrical response, second generation biosensors functions involving specific ‘mediators’ between the reaction and the transducer for generating improved response, and in third generation biosensors the reaction itself causes the response and no product or mediator diffusion is directly involved.

Components of NanobiosensorsA typical nanobiosensor comprises of 3 components; biologically sensitized elements (probe), transducer and detector

� The biologically sensitized elements (probe) including receptors, enzymes, antibodies, nucleic acids, molecular imprints, lectins, tissue, microorganisms, organelles etc., which are either a biologically derived material or bio-mimic component that receives signals from the analytes (sample) of interest and transmits it to transducer. And such nano-receptor may play a vital role in the development of future nanobiosensors.

� The transducer acts as an interface, measuring the physical change that occurs with the reaction at the bio- receptor/sensitive biological element then transforming that energy into measurable electrical output. Depending on the mode of action transducers may be classified into following categories and discussed in details.

� The detector element traps the signals from the transducer, which are then passed to a microprocessor where they are amplified and analyzed; the data is then transferred to user friendly output and displayed/stored.

Characteristics for an Ideal Nanobiosensor � Highly specific for the purpose of the analyses

i.e. a sensor must be able to distinguish between analyte and any “other” material.

� Stable under normal storage conditions. � Specific interaction between analytes should be

independent of any physical parameters such as stirring, pH and temperature.

� Reaction time should be minimal. � The responses obtained should be accurate,

precise, reproducible and linear over the useful analytical range and also be free from electrical noise.

� The nanobiosensor must be tiny, biocompatible, non-toxic and non-antigenic.

� Should be cheap, portable and capable of being used by semi-skilled operators

Role of Nanobiosensor in Agriculture

Nanobiosensors to detect soil quality and nutrientsProtection of the soil health and the environment requires the rapid, sensitive detection of pollutants and pathogens with molecular precision. Nano-particles are mini laboratories have the potential to precisely monitor temporal and seasonal changes in the soil-plant system. Soil solution can be allowed to react with nano-sensors that will give accurate measurement of availability of nutrients, moisture and physiological status of plants in the soils, to achieve the mission of precision agriculture.

Nano-Biosensors for Seed StorageSeeds during storage emit several volatile aldehydes that determine the degree of ageing. These gases are harmful to even other seeds. Such volatile aldehydes can be detected and seeds showing signs of deterioration can be separated and invigorated prior to their use. Electronic nose (E-Nose) can be used for it.

As an agent to promote sustainable agricultureTo promote sustainable agriculture nanofertilizers can be used. A nanofertilizer refers to a product that delivers nutrients to crops encapsulated within a nanoparticle. Nanofertilizers could be used to reduce nitrogen loss due to leaching, emissions, and long-term assimilation by soil microorganisms. Recently carbon nanotubes were shown to penetrate tomato seeds and zinc oxide nanoparticles were shown to enter the root tissue of ryegrass. This suggests that new nutrient delivery systems that exploit the nanoscale porous domains on plant surfaces can be developed. But, the nanofertilizers should show sustained release of nutrients on-demand while preventing them from prematurely converting into chemical/gaseous forms that cannot be absorbed by plants. To achieve this, biosensor could be attached to this nanofertilizer that allows selective nitrogen release linked to time, environmental and soil nutrient condition.

Zeolites are naturally occurring crystalline aluminum silicates that can a) enable better plant growth; b) improve the efficiency and value of fertilizer; c) improve water infiltration and retention; d) improves yield; e) retain nutrients for use by plants; f) improve long term soil quality and g) reduce loss of nutrients in soil. Zeolite holds nutrients in the root zone for plants to use when required. This leads to more efficient use of N and K fertilizers—either less fertilizer for the same yield or the same amount of fertilizer lasting longer



and producing higher yields.

Metal nanoparticle-based DNA-detection methodsThere are several nanosensors like ssDNA-CNTs probes/ optical biosensors to detect specific kinds of DNA oligonucleotides, MWNTs/ZnO/CHIT composite film modified GCE for immobilization ssDNA probes to effectively discriminate different DNA sequences. Two sets of gold nanoparticles are functionalized with two different sequences of thiol modified DNA that is complementary to the target DNA. Thus, in the presence of complementary target DNA, DNA-modified nanoparticles are connected into aggregates by duplex formation, which results in the shift of the characteristic surface plasmon band of gold nanoparticles from 520 to 570 nm, turning the red colour of colloidal gold to purple.

As a Device to Detect Contaminants and Other MoleculesSeveral nanobiosensors are designed to detect contaminants, pests, nutrient content, and plant stress due to drought, temperature, or pressure. Organophosphorus pesticides such as dichlorvos and paraoxon, at very low levels could be monitored by liposome-based biosensors. Zhang et al. developed a method for the detection of Escherichia coli (E. coli) using bismuth nanofilm modified GCE based on the principle of flow injection analysis (FIA).

Due to their unique characteristics and flexibility, nanobiosensors show great promise for sustainable agriculture. New prospects for integrating nanotechnologies into nanobio-sensors should be explored, cognizant of any potential risk to the environment or to human health. With targeted efforts by governments and academics in developing such enabled agro-products, we believe that nanotechnology will be transformative in the field of agriculture by focused research and development toward the goals for reaching sustainable agriculture.

20148

12. Role of Myconanoparticles in Phytopathogens ManagementARJUNSINH RATHAVA1, PUJA PANDEY2 AND RANGANATH SWAMY3

1& 3Assistant Professor, College of Agriculture, Jabugam, AAU, Anand 2Assistant Professor, B. A. College of Agriculture, AAU, Anand

IntroductionMyconanotechnology is an emerging field, where fungi can be used for the synthesis of nanostructures with desirable shape and size. myconanotechnology provide exciting waves of transformation in agriculture and fascinate researchers to contribute in providing continuous solutions through green chemistry approaches to facilitate food security. Nanotechnology is a field of applied science and cutting-edge technology that uses the physicochemical properties of the bulk molecules as a means to minimize and control their size, shape, and surface area, in order to produce different nanosized materials with unique physical, chemical, and biological properties.

There are many important aspect like enhancement of nutrients absorption by plants, protection of vegetation and nano formulated food ingredients, However, protection of plants with metal-based nanomaterial having a much greater surface area to volume ratio and unique antimicrobial agents compared to their bulk materials constitute one of the best applied solution offering successful results in controlling pathogens particularly bacterial and viral plant microbes.

Synthesis of MyconanoparticlesThe use of microbial cells for the synthesis of nanosized particles has emerged as a new technique for the coalescence of metal NPs. Several fungal strains have been used as promising resources for nanoparticle like Fusarium, Aspergillus, Verticillium and Pencillium. Different fungal species are efficient for production of metal NPs both intra as well as extracellular.

Fungi have a number of advantages for NP synthesis in relation to other microbes and plant products. The use of fungi in the forming of NPs is probably crucial since they produce large quantities of enzymes and are easy to grasp in the laboratory. Fungal mycelia can prevail against flow pressure and other circumstances in bioreactors and other chambers as compared to plant material.

Silver Nanoparticles as Antimicrobial AgentsMicrobes interact with nanomaterials and yield nanostructured materials because of their superlative performance, selective adsorption of metal ions, operation over a wide range of eco-friendly conditions, low cost, free possibility, transformation and high biosorption ability and the fact that large quantities can be obtained.



The emergence of nanoscience and nanotechnology in the last decade facil1itate opportunities for inspecting the antimicrobial effects of metal NPs. Nanoparticles have wide range of applications and are used in a number of fields, including medicine, pharmacology, environmental monitoring, electronics and agriculture.

The Applications of Nanotechnology in Plant Diseases ManagementPlant pathologists are engaged to protect food and

agriculture products pathogenic microbes like bacteria, fungal and viral agents and conserve the environment. A number of nanotechnologies can boost crop control in short to medium term. Nanomaterials are being cultivated that offer the probability to supervise pesticides, herbicides and fertilizers more accurately and safely by controlling precisely when and where they are liberated. Different types of nanomaterials such as copper, zinc, titanium, magnesium, gold, alginate and silver have also been tested, but Ag NPs have verified to be most effective as they have excellent antimicrobial potency against bacteria, viruses and fungi.

ReferencesKashyap, P. L., Kumar, S., Shrivastava, A. K and

Sharma, A. K. (2013). Myconanotechnology in agriculture: a perspective. World Journal ofMicrobiological Biotechnology.29:191–207.

Prasad, R., Kumar, V. and Prasad, K. S. (2014). “Nanotechnology in sustainable agriculture: present concerns and future aspects,” African Journal of Biotechnology. 13(6), pp. 705–713.

Mousa, A. A, Hassan, A, Mahindra, R, Ernest S. G. and Kamel A. (2015). Myconanoparticles: synthesis and their role in phytopathogens management. Biotechnology & Biotechnological Equipment,29(2) 221-236.

Salata, O. V. (2004). Applications of nanoparticles in biology and medicine. Journal of Nanobiotechnology. 2:3

AGRONOMY

20091

13. Bio-Fortification:ANovelApproachtoTackle Hidden Hunger and MalnutritionMANJANAGOUDA S. SANNAGOUDAR, AVIJIT GHOSH, KEERTHI, M. C. AND HANAMANT M HALLI

Scientist, ICAR-Indian Grassland and Fodder Research Institute, Jhansi-284003

IntroductionThe word Bio-fortification derived from Greek word “bios” means “life” and Latin word “fortificare” means “make strong”. Bio-fortification is the process of breeding food crops that are rich in bio-available micronutrients, such as vitamin A, zinc, and iron. Application of fertilizers to soil and/or foliar to improving grain nutrient concentration and the potential of nutrient containing fertilizers for increasing nutrient concentration of cereal grains. These crops are “biofortified” by loading higher levels of minerals and vitamins in their seeds and roots during growth. These crops can naturally reduce anemia, cognitive impairment, and other malnutrition-related health

problems that affect billions of people.Malnutrition is the most significant cause of global

mortality, with more than 50 per cent of diet-related deaths. Micronutrient deficiencies, especially Fe, Zn, Se, I and various vitamins, are common worldwide, affecting well over half of the world’s population, and multiple deficiencies often occur together.

Bio-fortification includes the production of micronutrient-dense food crops. Plant breeding strategies are very effective in this cycle because of their immense potential to improve dietary efficiency. Well-known examples of bio-fortification in the fight against micronutrient malnutrition are golden rice and low phytate legumes and grains. Application of fertilizers to soil and/or foliar to improving grain nutrient



concentration and the potential of nutrient containing fertilizers for increasing nutrient concentration of cereal grains.

Increasing the concentration of Zn and Fe in food crops, resulting in improved crop production and improved human health, is a significant global challenge. Micronutrient deficiency of Zn and Fe occurs in both crops and humans. Zinc deficiency is widely known as a significant risk factor for human health and cause of death worldwide.

Why we Need Bio-Fortification…? � Human requires at least 49 nutrients to meet their

needs � Primary source Agricultural products � Agriculture – fail to provide micronutrients in

developing & under developed countries � Cereals dominate the diet – poor in micronutrients,

vitamins, etc., � Over 4 billion people affected with micronutrient

malnutrition in all over the world � Food fortification couldn’t deliver the desired

result

Advantages of Bio-Fortification1. Since staple foods predominate in poor people’s

diets, this policy indirectly targets low-income households.

2. Once in operation, the network of bio-fortified crops is highly sustainable. Better varieties will continue to be grown and eaten year after year, even though the focus of the government and foreign support for micronutrient issues fades.

3. Bio-fortification provides a feasible means of targeting undernourished communities in increasingly remote rural areas.

Micronutrient Malnutrition Causes…. � More critical illnesses � Maternal deaths � Poor cognitive development � Growth affected � Work efficiency get affected. � Higher population growth rates.

Approaches of Bio-Fortification � Agronomic approach � Breeding or biotechnological approach

Genetic or Biotechnological Approach � Adequate genetic diversity occurs in the main

germ plasm banks to justify selection. � Micronutrient density characteristics are stable

across ecosystems. � It is possible to combine high micronutrient

density with high yield � Genetic control is easy enough to make breeding

economical. � Multiple limiting micronutrients can be enhanced

together.

Strategies for Bio-Fortification � Increasing the concentration of nutrients: Amino

acids, β carotene & Iron � Lowering the dietary anti nutrients: Iron & Zinc � Enhancing the dietary promoters: Iron & Zinc

Agronomic Bio-Fortification � Considered as short-term solution � Safe and accurate application systems to process

the fertilizers � Flexible; can be used for all crop species and

cultivars � Fast and cheap

Agronomic Approaches to Bio-Fortification � Balanced and adequate fertilization � Method of fertilization � Time of fertilizer application � Integrated nutrient management � Practicing crop rotation � Adoption of suitable Intercropping

Some of the Genetically Bio-Fortified Crops1. Golden rice

a) Created by Prof. Ingo Potrykus and Peter Beyer.

b) GR contains genes required to activate biochemical pathway leading to the accumulation of β Carotene.

c) Approach – Agrobacterium mediated.1. Iron bio-fortified rice

a) Iron biofortified rice was produced using three transgenic approaches.

b) Enhancing of Fe storage (SoyferH2), translocation (HvNAS1) and Fe flux into the endosperm (OsYSL2).

c) Ferrritn-ubiquitous protien for Fe storage and stores about 4000 Fe atoms in a complex.

d) Nicotinamine- Chelator of metal cations Fe(II) and Zn (II)

2. Quality protein maize (QPM)a) Amino acids: 55% more tryptophan, 30%

more Lysine, 38% less Leucineb) Biological Value – 80% (45%)c) Higher carotene & calcium utilizationd) All India Coordinated Maize Improvement

Project released varities: Shakthi, Rattan, Protina

e) Orange” Maize Could Save Eyesight of Millions of African Children

f) First Vitamin A-rich, Open-pollinated Maize Varieties Released



20093

14. Controlled Release Fertilizer (CRF): New Approach for Nutrient ManagementDR. VIJAY KUMAR DIDAL1*, DR. BRIJBHOOSHAN2, DR. SHALINI3, DR. KRISHNA CHAITANYA1 AND DR. RAJENDRAGOUDA PATIL1

1Assistant Professor, School of Agricultural Sciences and Technology (SAST), SVKM’S, NMIMS, Shirpur, Maharashtra-425405 2Auction Superintendent, Tobacco Board, APF No.5, Periyapatna, Mysore, Karnataka-571107 3Subject Matter Specialist, KVK (BUAT), Hamirpur, Uttar Pradesh-210505 *Corresponding Author Email: [email protected]

AbstractEveryone needs good nutrition for good and healthy life whether it is human being, livestock or plants. Higher yield of a crop depends upon supply of nutrients to the plant as per their nutritional requirement. Many crops are facing the problem of nutrient deficiency due to inability of the crop plants to uptake nutrients in desired quantity at the time of requirement which affect the yield of the crop negatively. The problem of nutrient deficiency occurs due to many reasons viz., deficiency of nutrients, faster release of nutrients results in availability of nutrients for very short period of time and loss of nutrients in the form of soil fixation, leaching, volatilization and denitrification. To overcome these problems, use of controlled release fertilizers is a better option to the farmers for proper nutrient supply to the crops for a longer period of time. A controlled-release fertiliser (CRF) is a coated fertilizer that releases nutrient slowly into the soil (i.e., with a controlled release period). In this article, we have discussed about controlled released fertilizers and their role in crop production.

Key words: Controlled release fertilizers, polymer coated fertilizers

IntroductionCrops have higher genetical yield potential but the exploitation of the yield potential of a crop in the field depends on many crop management factors. We can’t exploit their full yield potential until provide well prepared land, good quality seed, proper water management, nutrient management and pest management. Nutrient management is one of the basic crop management factors needed for successful crop production. Major lacuna in nutrient management is unavailability of nutrients for longer period of time due to faster release of nutrients resulted in loss of nutrients as happened in case of conventional or simple fertilizers. So, there is need of development of such fertilizers which release nutrient for longer period according crops demand.

Controlled release fertilizers (CRF’s) are such type of coated fertilizers that release nutrients for longer period or over an extended period of time at a rate driven primarily by prevalent temperature

and amount of available moisture of the root zone. Polymer coated fertilizers (PCF’s) are also a type of CRF’s, which are coated with various types of polymers (plastics). Fertilizer use efficiency is higher in case of controlled release or polymer coated fertilizer as compared to common fertilizers or uncoated fertilizers due to extended availability of nutrients throughout the life cycle of crop, very less nutrient losses, required in less quantity and production of more biomass and economic yield. Release of nutrients from controlled release fertilizers depends on coating thickness, chemistry or type of material used for coating, prevalent temperature and amount of available moisture in the soil.

Types of Coating Materials used for CRFsa) Polymer coatingb) Sulphur coating, andc) Sulphur plus polymer coating

Factors affecting Nutrient Release Rate from Controlled Release Fertilizers

� Temperature � Moisture � Size � Coating thickness � Coating failure (cracks, abrasion) � Coating material

Why to use Controlled Release FertilizersConventional fertilizers or simple fertilizers have several drawbacks. Some of them are listed below:

� Less efficient. � Faster release of nutrients makes availability of

nutrients for shorter period. � Faster release makes more susceptible to loss. � Required in large or huge amount. � Required in multiples split doses. � Increased cost of cultivation.

Due to the drawback and limitations of conventional fertilizers or simple fertilizers, controlled release fertilizers are best option for proper nutrient management because of above mentioned drawbacks of conventional fertilizers.



Advantages of Controlled Release Fertilizers � Increased nutrient release timing � Meet plant demand timely and efficiently � To improve the yield and reduce the cost of

production � Reduction of plant toxicity � Reduction in ground water pollution and water

bodies � Fertilizers are released at a slower rate throughout

the season; so that plants could take up most of the nutrients without much waste.

Limitations for use of Controlled Released Fertilizers

� Applying sulphur coated urea almost always lowers soil pH by leaving acidic residues on application..

� Less popular among farmers � Expensive

Examples1. Nitrogen fertilizers:

a) Polymer Coated Urea (41-0-0, 44-0-0)b) Polyon (40% N)

2. Phosphorus fertilizers:

a) AVAIL (Phosphate enhancer)3. Potassium fertilizers:

a) POLYON-45 (0-0-45)4. Sulphur fertilizers:

a) PCSU (Polymer coated sulphur coated urea): (39-0-0, 41-0-0, 43-0-0), 11-15% sulphur

b) Poly-S5. Micronutrients fertilizer:

a) FeSO4 coated fertilizer

b) MnSO4 coated fertilizer

c) ZnSO4 coated fertilizer

SummaryControlled release fertilizers are mostly use for lawns, vegetables, horticulture crops and high value crops which are more efficient in nutrient management than conventional fertilizers. Controlled release fertilizers are available for most the nutrients (Macro and micro nutrients) and increase the efficiency of all nutrients by reducing the loss of nutrients by controlled supply of nutrients with a controlled release period. Their cost is higher than conventional fertilizers but required in smaller quantity than conventional fertilizers or uncoated fertilizers.

20100

15. Organic Farming: A Review in Indian ContextAJIT U. MASURKAR AND AKASH D. LEWADE1B.Sc. Agriculture (4rth Year) R.B.C.A. Pipri, Wardha 2Assistant Professor, Section of Agronomy, R.B.C.A. Pipri, Wardha

Sustainability is the basis of livelihood for humans. With advances in technology, we have achieved the record production as well as productivity in agriculture, but no one is sure about quality of produce as because of uncontrolled use of pollutable inputs. It is much more discussed about organic way of producing crops in today’s agriculture production system. As we all know that organic farming is the production system that largely avoids use of chemically synthesized inputs. But it is not restricted up to that; in broad sense, we can say that, Organic farming is the food production system that might be considered as a holistic approach which utilizes all biodegradable, recyclable, non-polluting eco-friendly inputs in order to build-up soil-fertility & productivity, reduce insect-pest-diseases outbreak, minimize the contamination of food chain, restrict the over-exploitation of natural resources and ultimately to bring-up sustainability in crop-production and livelihood of dependent community. Crop production by organic way is a tricky and risky task. As mentioned earlier, it is a holistic approach that comprises of:1. Crop diversification2. Soil fertility management through organic manure

& fertilizers3. Non-Chemical weed control

4. Biological insect-pest and disease management.Organic farming does not only mean of replacing

synthetic fertilizers and other chemical inputs with organic inputs and other bio-formulations but it also emphasized on long term soil health and fertility management through a comprehensive way. Organic is not any new invention to Indian farming; in spite it is an old age production system followed since centuries in India. Use of animal dung as manure is mentioned in Rigveda, Atharva-veda signifies the importance of green manure whereas kautilya in his Arthashastra (321-296 BC) underlines that manures like oil cake, animal excreta etc. Krishi Parashara (10th century) states that, “life of farmers is solely dependant upon the microbes present in the soil.” Various practices related to Organic crop production has been described in numerous Indian literatures viz. Panini’s Astadhyayi, Patanjali’s Mahabhashya, Varahamihira’s Brihat Samhita etc. All the wishdom and knowledge gained a practice used by Indian Farmers were pass in the Form of ITK from generation to generation. Even today, the myths, the belief system, the rituals and festivals of Indian Culture enlighten various principles of soil, plant & animal health, biodiversity. Sir Albert Howard, a principal figure in early organic movement of India is



considered as the Father of Modern Organic farming. He developed and documented various techniques in organic farming. His book, An Agricultural Testament contains bulky material on Organics. The traditional knowledge of farming without harming ecosystem has been preserved by some tribes in the hilly forest and that’s why the practice of organic cultivation in such areas are categorized as “Organic by default.” Sikkim has been declared as the first Organic state by government of India under this consequence. The various Organic approaches adopted in Indian context are as follows:

Bulky Organic ManureThese are the organic sources of plant nutrients that have to be applied in bulk. It includes FYM, compost, vermicompost, sewage, sludge, bio-gas slurry, sheep manure, poultry manure etc.

FymA decomposed mixture of cow dung, urine, litter and left-over feed material. It contains 0.5:0.3:0.5% NPK.

CompostA well decomposed organic manure prepared from bo-degradable waste by the activity of microbes.

� Farm compost - 0.5:0.15:0.5% NPK � Town compost – 1.4:1.0:1.4% NPK

Vermi-compostIt is well decomposed organic manure prepared from waste material by the activity of earthworms.

Night soilHuman excreta both solid and liquid is night soil. It contains 1.2-1.3:0.8-1.0:0.4-0.5% NPK.

Sheep and goat manureIt contains 3.0:1.0:2.0 % of NPK.

Poultry manureIt contains 3.03:2.63:1.40 % NPK.

Concentrated Organic ManuresThese are the nutrient sources of organic origin which are relatively high in nutrient content and has to be applied in relatively smaller quantities.

Bird guanoIt contains 7.8:11-14:2-3% NPK.

Raw bone mealIt contains 3-4:20-25 % N & P.

Oilcakes � Castor cake contains 5.5-5.8:1.8-1.9:1.1 % NPK � Karanj cake’s NPK content – 3.9-4:0.9-0.1:1.3-

1.4% � Mahuacakes’s NPK content – 2.5-2.6:1.8-1.9:1.8-

1.9% � Groundnut cake’s NPK content – 7-7.2:1.5-1.6:1.3-

1.4% � Undecorticated cotton seed contains- 3.9-4:1.8-

1.9:1.6-1.7 %NPK � Sesamum cake contains: 6.2-6.3:2-2.1:1.2-1.3%

NPK.

Green manuringThe process of turning and ploughing into the soil, the undecomposed green plant tissues is known as green manuring. Plants like Sunhemp, Dhaincha, Green gram, Cowpeea are used as an as green manure. On an average, green manuring gives 60-80 kg Nitrogen per hectare.

Growth promotersVarious growth promoters of organic origin have proved their practical utilities in boosting up the productivity of cropping systems. Some of them are:

JeewamrutOne of the cheapest organic produce prepared by fermenting cowdung and urine alongwith pulse flour and jaggery that is used on crops.

AmritpaniA natural microbe enriched produced by incubating cow products with honey, that acts as an effective tool for enhancing germination and managing soil borne diseases.

PanchagavyaAs the name indicates, it is the combination of five cow products i.e. cowdung, cow urine, cow milk, cow’s curd and cow’s ghee alongwith jaggary, ripened banana & tender coconut water that plays a significant role in stimulating crop growth.

VermiwashA liquid organic manure produced by the earthworms during the preparation of vermicompost.

Humic acidIt is one of the humic substances obtained during process of organic matter decomposition with medium molecular weight and alkali soluble nature. It is a group of molecules that bind to and help plants roots receive, water and nutrients.

BiofertilizersIt is the microbial preparation of live and viable cells capable of solubelizing, mobilizing and converting unavailable forms of nutrients to available ones. Various types of biofertilizers are used for various crops like Rhizobium for Legumes, Azotobacter for rice, Azospirillum for sorghum, Azolla for rice along with this PSB, VAM etc. are also used in various crops. Rhizobium can fix 50-200 kg N/ha per year. BGA can add up to about 20-25 kg N/ha to rice fields.

Cultural and Mechanical Weed ControlFor an effective control of weeds without use of



chemical, use of agronomic and mechanical measures is common. Agronomic manipulations in forms of adjusting seed rate, sowing dates, sowing depth and method, plant spacing, water & nutrient management are commonly adopted in Organic farming. Whereas mechanical measures like weeding, hoeing, sickling, mowing, chaining, dredging, buring are some commonly followed methods in organics.

Bio-HerbicidesThese are the preparations containing live cells or latent microbes that are capable of killing weeds through various modes. Devine, Collego, Biolophos are

some bioherbicides. Alongwith this some insects like Mexican bettle, Crosidosema lantana are effective in weed control.

Plant ProtectantsVarious plants protectors of plant origin like Need Seed Kernal Extract (NSKE), Neemastra, Dashparniarka, Agni-astra, Tobacco extract, Chilli-Ginger-Garlic extract etc. are common in Organic farming.

Reference1. Handbook of ICAR.2. Fundamentals of agriculture- vol.1 - Arun Katyayan.

20110

16. Silicon: An Element to Recover Stress Tolerance in PlantsMAHESH GURAV AND RUSHIKESH PAWAR

Department of Agronomy, College of Agriculture, Dhule (Mahatma Phule Krishi Vidyapeeth, Rahuri 413722) *Corresponding Author Email: [email protected]

IntroductionSilicon (Si) is a non-essential nutrient for most of the plants. However, in field crops it is known to affect plant growth and quality, photosynthesis, transpiration and enhance plant resistance to stresses such as drought. The beneficial effects of Silicon are associated with its high deposition in the plant tissues, enhancing their rigidity and strength. Silicon also enhances the host resistance in plant against diseases by regulating the defence reaction mechanism. Many plants cannot accumulate Silicon at enough levels that it can be beneficial. So, manipulating the root uptake capacity for Silicon can improve the ability of the plants to overcome biotic and abiotic stress.

History of Silicon in PlantsSilicon dioxide comprises about 50-65 % of soil mass and is regarded as the second most abundant element found in the soil after Oxygen. The role of Silicon in plant growth was overlooked till the 20th century. Due to its abundant availability in nature and the symptoms of Silicon toxicity or deficiency being non apparent, Silicon was overlooked for its role in regulating the physiology in plants.

Over the period, continuous cropping and continuous use of synthetic fertilizers comprising Nitrogen, Phosphorus and Potassium have depleted the level of Silicon in the soil that can be made available to the plants. The deficiency of Silicon in soil is now considered as a limiting factor fin crop production. Silicon till today is not recognized as an essential element for plant growth and development, but the beneficial effects of the element on plant growth, development, yield, disease and stress resistance have been observed.

Silicon in PlantsSilicon is recognized as a plant nutrient anomaly because it is presumably not essential for plant growth and development. But soluble Silicon has been found with results depicting increase in growth and development of crops like rice, sugarcane, cucumber, pomegranate and other cereal crops.

The accumulation of Silicon has been reported as 0.5% to 1.5% in Sugarcane, while wetland rice absorbs silicon in the form of monosilicic acid Si(OH)

4,

equivalent to 4.6% to 6.9% of the dry matter of rice.

Role of Silicon in Stress ResistanceSilicon is considered as a beneficial element and has the ability to protect the plants from multiple biotic and abiotic stresses; hence Silicon is considered as evident under different stress conditions.

Silicon has been found effective in controlling various fungal and bacterial diseases in plants. Silicon improves the resistance in rice to leaf and neck blast, brown spot, stem rot and sheath blight. Silicon also decreases the incidence of powdery mildew in cucumber, rust in cowpea and ring spot in sugarcane.

Two mechanisms for Silicon enhanced resistance to diseases have been proposed. One of the mechanism states that Silicon acts as a physical barrier and is deposited beneath the cuticle and forms a cuticle-Si double layer. The layer formed impedes the penetration by fungi and acts as barrier to fungal infections. Another mechanism proposed is that Silicon acts as a modulator of host resistance to the pathogens causing disease in plants.

Silicon also ease many abiotic stresses like the salt tress, metal toxicity, imbalance of nutrients and physical stress like lodging, drought, radiation,



freezing, high temperature.Silicon promotes cell elongation but do not

participate in cell division. Plants that accumulate Silicon in large amount has benefit that the plants enhance stress resistance. To derive beneficial effects of Silicon the plant must acquire the element in larger concentrations.

Future ProspectsThe accumulation of Silicon in plants is controlled by the ability of the roots to uptake Silicon. Silicon uptake being a complicated process is manipulated by multiple genes. Furthermore, dicots plant has less ability to accumulate Silicon; hence manipulating by genetic improvement the roots may be able to accumulate more Silicon and hence can overcome different biotic and abiotic stresses.

20111

17. Direct Seeded Rice: A Resource-Saving Technology forProfitableRiceProductioninIndiaMANOJ K. N.

Ph.D. Scholar, Department of Agronomy, UAS, GKVK, Bengaluru-560065

Rice is the dominant staple food of about 4 billion people worldwide providing about 33 per cent of the total caloric intake of most Asians. The global rice demand will continue to increase from 479 million tons (milled rice) in 2014–15 to 544 million tons in 2029-30. Whereas in India it feeds the belly of more than 800 million people and has a share of 41.39 per cent in total food grain production and 55 per cent of cereals production in the country, contributing 20-25 per cent to Agricultural GDP. In India, rice is grown on nearly 43.77 m ha with total production of 112.79 million tons and productivity of 2576 kg/ha. Given the projected population growth, it is estimated that the annual rice yield growth in the next 10 years will have to increase to around 1.2 to 1.5 per cent from its current rate of less than 1 per cent. About 55 per cent of the total rice area is irrigated and concerns are increasing about the availability of water for crops due to competition with urban areas and most of the areas in South and Southeast Asia may suffer from physical or economic water scarcity. On the other hand, other factors like inefficient use of inputs (fertilizer, water, labour), less availability of labour, climate change and rising cost of cultivation are major hindrance for achieving the target yield.

In most of the Asian countries including India, rice is grown by manual transplanting of seedlings into puddle soil with 20-30 days old seedlings by rising in nursery. Moreover, puddling requires additional water (~200 mm) which is becoming scarce day by day and also labor becoming costlier each day. In India per capita water availability decreased by 68.8% between 1951 and 2001 (5831 m3 and 1820 m3, respectively) and is likely to decline by 80.4% by 2050 (1140 m3). Hence at present situation, decreased availability of water and labor and increased production costs are associated with the transplanting rice resulting in paddy production less profitable to the farmers. All these above factors demand a major shift from conventionally tilled- puddle transplanted rice (CT-

TPR) to direct seeding of rice (DSR) as it saves water up to 25%, energy up to 27%, labor up to 35-40 man days per hectare with additional advantage of reduced methane emissions and early maturity of crop by 7-10 days.

There are mainly three kinds of direct-seeded rice systems: dry-seeded (Dry-DSR), wet-seeded (Wet-DSR) and water-seeded. In Dry-DSR, rice is established using several different methods, including (1) broadcasting of dry seeds on unpuddled soil after either zero tillage or conventional tillage, (2) dibbled method in a well-prepared field, and (3) drilling of seeds in rows after conventional tillage, reduced tillage using a power- tiller-operated seeder, zero tillage, or raised beds. It is usually practiced in rainfed upland ecosystems of the Asian countries where water is scarce. In contrast to Dry-DSR, Wet-DSR involves sowing of pregerminated seeds with a radical varying in size from 1 to 3 mm on or into puddle soil. When pregerminated seeds are sown on the surface of puddled soil, the seed environment is mostly aerobic and this is known as aerobic Wet-DSR. When pregerminated seeds are sown or drilled into puddled soil, the seed environment is mostly anaerobic and this is known as anaerobic Wet-DSR. In both aerobic and anaerobic Wet-DSR, seeds are either broadcast or sown in-line using a drum seeder or an anaerobic seeder with a furrow opener and closer. It is usually practiced in Srilanka, Vietnam, Malaysia, Thailand and India where labor is scarce. However, water seeding has gained popularity in areas where red rice or weedy rice is becoming a severe problem and it is most common seeding method used in California (United States), Australia and European countries to suppress difficult-to-control weeds, including weedy rice. This method is also becoming popular in Malaysia. In this method, pregerminated seeds (24-h soaking and 24-h incubation) are broadcast in standing water on puddled (Wet-water seeding) or un-puddled soil (Dry water seeding). Normally, seeds, because of



their relatively heavy weight, sink in standing water, allowing good anchorage. The rice varieties that are used possess good tolerance of a low level of dissolved oxygen, low light, and other stress environments. In addition to irrigated areas, water seeding is practiced in areas where early flooding occurs and water cannot be drained from the fields.

Besides many advantages, the direct seeded rice

systems are not without critics. Weeds are the top most biological constraint to the production as well as adoption of direct-seeded rice systems. Hence, if we succeed in reducing the risk of yield loss due to weeds then DSR will be the best viable technology for rice production in near future for feeding the ever-increasing population.

20131

18. Importance of Panchgavya in Crop Production*SUNIL AND SEEMA DAHIYA

College of Agriculture, CCS Haryana Agricultural University, Hisar – 125 004, Haryana, (India) *Corresponding Author Email: [email protected]

Panchgavya is an organic source of nutrients made up of five different cow products and applied to crop plants since Vedas. It is helpful in improving the efficiency of plants by creating a favourable environment for microorganisms present in soil which improves the physical and chemical properties of soil. The main objective of panchgavya application is to enhance yield potential of crops. It can be applied in form of foliar spray, soil application or seed treatment.

IntroductionPanchgavya is known as excellent source of nutrients from the time of Vedas (divine scripts of Indian wisdom). Panchgavya is made up of two words- panch and gavya, panch means five and gavya means products. According to its description in Ayurveda, it is prepared from mixing of five cow products viz. milk, curd, ghee, urine and dung. Panchgavya is mainly used in organic farming, but its use along with fertilizers and pesticides in inorganic farming is also mentioned in our literature. Panchgavya enhances the process of mineralization by increasing the activity of soil microbes. As it is organic source of nutrient, so it has long lasting effect on soil fertility. Due to continuous practicing of inorganic farming since a longer period of time, several hazards for human, animal and bird health has arisen. Environment quality has also degraded due to indiscriminate use of chemicals in inorganic farming. By considering these adverse impacts, it becomes essential for us to shift towards the organic farming. Panchgavya has a very pivotal role in organic farming to meet the nutrient requirement of crops. It is mainly applied in form of foliar spray with 3% concentration using hand operated sprayer having high pore sized nozzle.

Quantities of Ingredients of Panchgavya PreparationIngredients Quantity Ingredients QuantityCow’s dung 5 kg Ghee from

cow’s butter1 litre

Ingredients Quantity Ingredients QuantityCow’s urine 3 litre Tender coconut

water3 litre

Cow’s milk 2 litre Curd from Cow’s milk

2 litre

Sugarcane juice 3 litre Banana 12 in number

Nutrient Status of PanchgavyaPanchgavya is a rich source of various macronutrients such as nitrogen, phosphorus, potassium and micronutrients such as iron, manganese, zinc, boron etc. which are the basis of growth and development of plants. It is also a good source of vitamins and plant growth regulators like auxin, gibberlins, cytokinin etc. Microorganisms such as Pseudomonas, Bacillus, Azotobactor, Phosphorus solubilizers etc are present in abundant quantity in panchgavya applied fields.

Beneficial Effects of Panchgavya Application

Effects on plants � Panchgavya applied field produces plants of

bigger leaves and dense canopy. � Biological efficiency of plants is enhanced by

synthesizing maximum metabolites and synthates. � More no. of side shoots is produced so that

maximum fruits are obtained at maturity time. � Plants produces dense and profused roots which

remain fresh for longer time and grows into deeper layer so that maximum uptake of nutrient and water takes place.

� Yield potential as well as quality of crops is improved.

� Cost of cultivation is reduced by replacing the expensive chemical fertilizers.

� Evaporation is also reduced by forming an oily film on leaves and stem.

Effects on soil propertiesMineralization process is enhanced due to favourable growth and development of microorganisms.

Soil fertility is improved due to increase in the



macro and micro nutrient content of soil by enhanced microbial activity of soil.

Water holding capacity is improved because of its richer content of organic matter.

Effects on insect pest and diseasesA plant becomes tolerant to insect pest and diseases by the application of panchgavya.

Various disease-causing pathogens are effectively controlled by the production of beneficial metabolites such as hydrogen peroxide, organic acids and antibiotics.

Effect of panchgavya on major cereal cropsRice

� Tillering, grain weight and cooking quality is increased.

� No. of chaffy grains and broken rice during milling is reduced.

� Harvest is advanced by 15 days.

Wheat, barley, maize and sorghum � Availability of nutrients to crop plants is increased

as a result of which growth of plants is increased to a great extent.

� Palatability is also improved to a greater extent. � Harvest is also advanced by 15 days.

Overall Advantages of Panchgavya � It is farmer and eco-friendly method to provide

balanced nutrition to crop plants. � It improves the soil health and fertility and

ultimately increases the quality and yield potential of crops.

� No special machinery and techniques are required for its application.

� Cost of cultivation is reduced as use of expensive chemical fertilizers, pesticides, fungicides and herbicides is restricted.

Constraints in Panchgavya Adoption � Lack of awareness and its slow acting nature � Enhanced weed infestation by its application. � Limited availability in market

ConclusionDue to large scale practicing of inorganic farming for a longer period of time, several hazards are produced for both human and animal health. Environment quality is also degraded due to indiscriminate use of chemicals. These hazards can only be minimized by reducing our overreliance on chemicals and shifting towards the organic farming. Panchgavya is best option for nutrient management in organic farming due to its greater efficacy and potential to increase yield levels of crops. It is not much popular among farmers due to lack of awareness. This problem can be solved by various effective extension activities and programs such as result demonstration and method demonstration.

20159

19. Major Constrain in Pulses Production and Productivity in IndiaGAURAV SHUKLA1* AND DURGESH KUMAR MAURYA2

1* 2 Ph.D. & M.Sc. Scholar, Department of Agronomy, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut-250110 *Corresponding Author Email: [email protected]

AbstractIndia is still big and vegetarian in dietary habits and depends on vegetation Sources to meet our daily protein requirement. India is bound to be the global leader in this matter since pulses production and consumers. India is an importer of pulses production Pulses/legume crops have been stable over the years. In addition to fix atmospheric ‘N’ in readily available form Upcoming successful harvest. It also contributes to maintaining the production system through the physical, chemical and biological improvement of soil properties, as a rotation effect. The seed replacement rate is still (<30%) lower than grain especially wheat and rice. This paper focus on constrains.

IntroductionPulses are leguminous crops which is an important

source of protein (20-25%), essential amino acids, vitamins and minerals. Pulses are used as “dal” in India. This is also known as “Poor man’s meat” and the rich man’s vegetable because pulses provide significant nutritional and health benefits, and are known to reduce several non-communicable diseases such as colon cancer and heart diseases (Yude et al, 1993; Jukanti et al, 2012). The leguminous crop can fix atmospheric nitrogen (30-150 kg ha-1). Pulses can grow in diverse climate conditions and any type of soil and play an important role in enhancing the soil fertility through nitrogen fixation, the release of soil-bound phosphorus, maintain cation exchange capacity (CEC) and increase the sustainability of the farming system. World’s largest producer and consumer of pulses in India. In India major pulses are grown like chickpea (Cicer arietinum), pigeon pea (Cajanus



cajan), pea (Pisum sativum var. arvense), black gram (Vigna mungo), green gram (Vigna radiate), lentil (lens esculenta), lablab bean (Lablab purpureus), horse gram (Dolichos uniflorus), moth bean (Vigna aconitifolia), khesari (Lathyrus sativus), broad bean (Vicia faba), and cowpea (Vigna unguiculata), however, chickpea, pigeon pea, green gram, black gram, and lentil are more popular among of these. Mostly pulses are grown in two seasons (1) Kharif (June-Oct), and (2) Rabi season (Oct-April). Pigeon pea, urdbean, mungbean, and cowpea are grown during the Kharif season, while Chickpea, lentil, and dry peas are grown in the Rabi season. Nutritional value of pulse-

Constituents MagnitudesCarbohydrate 55 – 60%Protein >20%Fibre 3.2%Fat >1.0%Vitamin A 430-489 IUVitamin C 10-15 mg/100 gCalorific value 343Calcium 69 -75mg/100gIron 7-10mg/100 gPhosphorus 300-500 mg/100 g

Major Constrains in PulsesFollowing constraints in pulses which decrease production and productivity of pulses

Agronomic constraintsAccording to Ramakrishna et al. (2000) and Reddy (2009) incorrect sowing time, low seed rate, faulty sowing method, insufficient irrigation, inadequate intercultural operation, sowing under utera without proper management practices are major agronomic constraints. Resultant upon delayed planting, early meet with severe cold, growth, and development of lentil crop gets disadvantaged for a considerable period. Ramakrishna et al. (2000) reported that subsequent plants get comparatively less time to complete their life cycle which, by and large forces maturity. One earlier study showed that the area under most pulses predetermined, but as the irrigated area grows, pulses are moved to rainy areas and their areas are changed by grain or some cash crops (Singh et al., 1995). In India, the irrigated area under pulses was only 12 percent, however the area under wheat and paddy about 60 percent of the total area (Reddy and Reddy, 2010).

Pests and diseasesAlthough legumes are susceptible to many insect pest and disease, if these are not controlled, pulses or legumes production decrease. Legumes crop plentiful affected by Fusarium wilt disease, which causes the losses of yield. Legumes grains is caused heavy damage by pests during storage. If proper crop rotation is followed legumes are in general pest-free crop under normal condition. Singh et al. (2013b) and Singh et al.

(2013g) reported that Pod borer, Aphids, and Wilt are major insects and disease pests.

Crop Disease Insect pestsChickpea Fusarium wilt,

Ascoochyta blight, botrytis grey mould, and stunt virus

Pod borer and cut worm

Pigeon pea Sterility mosaic virus, Fusarium wilt, Phytophthora stem blight Alternaria leaf spot and powdery mildew

Pod borer and pod fly leaf tier

Urd and Mung

Yellow mosaic virus, Cercosora leaf spot, powdery mildew, leaf crinple virus and root rot

white fly, Jassid and pod borer

Lentil Rust, wilt Sclerotinia blight, collor rot

Pod borer

Field pea Powdery mildew, rust, downy mildew, wilt

Pod borer, stem borer, leaf minor

Technological constraintsA legume is grown in the country under various agro-climatic conditions (soil type, rainfall, and thermal governance). The region calls for specific production techniques, including crop varieties with characteristics related to prevalent biological and abiotic stresses. Even the use of organic fertilizers and pesticides must be based on isolated strains from areas with similar agro-climatic conditions to be effective. Our research and development program in pulses is yet to be adequately appreciated and addressed. Production technology for a legume crops reliant on to be soil type, specific equally applicable for tillage, and seeding method (Singh et al. 2012).

Varietal constraintsAccording to Singh et al. (2013), and Ramakrishna et al. (2000) lack of low harvest index, high yielding varieties, high susceptibility to insect pests and disease, flower drops, absence of short duration varieties, intermediate growth traditions, poor reaction to inputs and Instabilities in shows are the few of the varietal constraints desires immediate attention.

ConclusionIt is concluded that the pulses are grown under good agronomic practice, control of insect pest disease, application of production technology to increase the production and productivity of pulses.

ReferenceJukanti AK, Gaur PM, Gowda CLL and Chibbar RN.

2012. Nutritional quality and health benefits of chickpea (Cicer arietinum L.): a review. British Journal of Nutrition 108, S11-S26.

Ramakrishna A, Gowda CLL and Johansen C.2000. Management factors affecting legumes production in the Indo-Gangetic Plain. In: Legumes in rice



and wheat cropping systems of the Indo-Gangetic Plain-constraints and opportunities. (Johansen C, Duxbury JM, Virmani SM, Gowda CLL. Eds.). pp. 156-165. ICRISAT, Patancheru, Andhra Pradesh.

Reddy AA and Reddy GP. 2010. Supply Side Constrains in Production of Pulses in India: A Case Study of Lentil. Agricultural Economics Research Revie. Vol. 23 January June 2010 pp 129-136

Reddy AA. 2009. Pulses Production Technology: Status and Way Forward. Economic & Political Weekly 44 (52): 73-80

Singh AK, Bhatt BP, Singh KM, Kumar Abhay, Manibhushan, Kumar Ujjawal, Chandra Naresh and Bharati RC 2013b. Dynamics of powdery mildew (Erysiphetrifolii) disease of lentil influenced by sulphur and zinc nutrition. Plant Pathology Journal12 (2):71-7.

Singh AK, Bhatt BP, Sundaram PK, Chndra N, Bharati RC and Patel SK. 2012a. Faba bean (Viciafaba L.)

phenology and performance in response to its seed size class and planting depth. Int. J. of Agril. & Stat. Sci. 8 (1): 97-109.

Singh AK, Singh KA, Bharati RC and Chadra N.2013e. Response of intercrops and nutrient management on the performance of tobacco based intercropping system and assessment of system sustainability. Bangladesh J. Bot. 42(2): 343-8.

Singh Deepak, Kumar Ashish, Singh Anil, Kumar and Tripathi, Hirdai Shankar. 2013g. Severity of chickpea wilt in north Bihar and nutritional studies on Fusarium oxysporum f. sp. Ciceri J. Plant Disease Sci. 8(2):137-40.

Singh KM and Singh RKP. 1995. An Economic Analysis of Lentil Cultivation in N-E Alluvial Plains of Bihar. Economic Affairs 40 (3):157-63.

Yude C, Kaiwei H, Fuji L, Jie Y. 1993. The potential and utilization prospects of kinds of wood fodder resources in Yunnan. Forestry Research 6, 346-350.

20162

20. Agronomic Weed Management: An Approach for Sustainable Crop ProductionHARSITA NAYAK

Ph.D Scholar, Department of Agronomy, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005 *Corresponding Author Email: [email protected]

Introduction: Weeds are very problematic as they affect the crop yield greatly. It is estimated that India loses crop produce worth $11b to weeds every year. In the year 2017-18, the actual economic losses due to weeds were found to be maximum in rice ($4.42b) followed by wheat ($3.376b) and soybean ($1.56b). In order to sustain the production and productivity they need to be managed properly from the early stage of their growth. There are several weed control strategies which are being incorporated by the farmers to keep the weed population below threshold level. However, in the recent years, agronomic weed control measure has gained popularity due to public awareness on health issues, environmental pollution and increased cost of production. Besides, rapid evolution of herbicide resistant weeds due to continuous and prolonged application of same herbicide or herbicides having similar mode of action has made us to shift from chemical weed control to cultural weed control methods.

Definition: Agronomic weed control measure generally includes non-chemical cultural practices that use the competitiveness of the crop to maximize its growth while diminishing the growth and subsequent competitiveness of associated weeds.

Types of Agronomic Weed Management1. Crop density: Crop competitiveness against

weeds can be enhanced by increasing crop

density. At higher crop density, increased crop canopy closure causes a greater reduction of interception of solar radiation to the soil surface and the weeds beneath the crop canopy, creating smothering effect. As a result, weed population, weed dynamics and weed seed production reduce significantly creating a suitable environment for crop.

2. Row spacing: Wider spacing favours weed invasion whereas narrow row spacing and high seeding rate improve the competitive effect of crops on weeds. Bidirectional sowing of crops could be an alternative as it provides minimum space for weed establishment. This also results in reduced dependence on herbicide application.

3. Sowing time: Alteration of sowing time determines the type and degree of weed infestation along with weed flora composition during the growing period. For example, early sowing results in lower intensity of Sorghum halepense, Panicum dichotomiflurum, Amaranthus retroflexus L., and Portulaca oleracea L. Crop sown at optimum time with adequate soil moisture and temperature, especially in dry region, will be more vigorous and suppressive over crops grown at adverse condition.

4. Use of competitive cultivars: Cultivars having ability to either tolerate the weed competition or suppress the weed growth can be best suitable



to control weeds effectively. Crops with vigorous growth habit, early ground cover, deep root system, early biomass production, rapid leaf area development and ability of efficient utilization of water and nutrients can be able to keep the weed population minimum. Varieties having allelopathic characteristics can be selected to reduce the crop damage by weeds.

5. Cover crops: Cover crops are grown between growing seasons on arable land in order to prevent soil erosion or to diminish weed population. Inclusion of cover crop on an uncropped land in a rotation has been proven to be beneficial because of its ability to suppress weeds while improving soil health.

6. Use of mulch: Mulching, is an important agronomic tool that not only reduces soil moisture evaporation but also helps in weed suppression. Mulch can be of different types viz., live or organic mulch, mulch from plastic or inorganic materials and mixed mulch. However, cost of mulching makes it economic only for high value perennial crops.

7. Crop rotation: Growing of different crops in succession in a piece of land in a year is known as crop rotation. Crop rotation results in changing

patterns of resource competition, soil disturbance, mechanical damage, allelopathic interference which ultimately lowers intensity of notorious crop associated weed infestation. It also changes the weed diversity, thus preventing establishment of the weed community that excellently fitted with the growing cycle of a particular crop.

8. Nitrogen fertilization: Crop-weed competition for nutrients, particularly for nitrogen, is a major problem since the availability of nitrogen is often the limiting factor in crop growth especially in soils with low nitrogen. Broadcasting of nitrogen generally favours weed growth whereas banding near seed row helps seedlings get nutrient more quickly increasing crop competitiveness. Similarly, applying nitrogen during spring rather than in fall lowers weed density and increases crop yield.Conclusion: Though agronomic practices solely

are not capable of managing weeds completely, but they may pose indirect adverse effects on weeds and suppress them to some degree. Thus, along with herbicide application, farmers should widen and modify their weed management practices by including these agronomic tools to reduce the risk of evolution of herbicide resistant weeds.

20169

21. Physiological and Biological Aspects of Herbicide Activity and SelectivityJAGADISH JENA1*, TWINKLE JENA2 AND SAMEER RANJAN MISRA3

1Department of Agronomy, Indira Gandhi Krishi Vishwavidyalaya, Raipur, Chhattisgarh 2Department of Agronomy, Institute of Agriculture Sciences, Banaras Hindu University, UP 3Division of Agricultural Chemicals, ICAR-IARI, New Delhi-110012, India. *Corresponding Author Email: [email protected]

IntroductionEfficiency of herbicide is the function of its selectivity and the selectivity of the herbicide is the function of its absorption, translocation and metabolism. Herbicide absorption and translocation must be effective and efficient for more efficacy of herbicide. For effective herbicide activity and its selectivity, one should know how herbicides move from site of application to site of action. Among systemic herbicides some are soil and some are foliage active; accordingly, they move inside the plant system. Mode of action of herbicide involves its absorption, translocation and metabolism inside the plant system. To increase the efficacy of herbicide the optimum rate of application is pre decided for different herbicides for different plant system which depends on some factors i.e. absorption of herbicide, translocation of herbicide and metabolism of herbicide.

Absorption of HerbicidesHerbicide absorption may be defined as the physical

entry of herbicide inside plant system. Soil active herbicides absorb mainly through passive uptake, whereas foliage active are absorbed mainly through active uptake. Factors affecting the absorption of soil active herbicides are i.e. Soil physical, chemical and biological factor, moisture and organic matter content of soil, rhizosphere environment, rooting pattern of plants/weeds and cell wall permeability of weed/crop seeds. Factors affecting the absorption of foliage active herbicides i.e. cuticle thickness, presence of trichomes or leaf hairs, cracks in cuticle, physiological state of the plant, orientation of leaf and age of the plant.

Herbicide TranslocationHerbicide translocation may be defined as the movement of absorbed herbicide from site of application to site of mechanism. It may be differentiated into; no or limited mobility in treated plant, symplastic movement, apoplastic movement and symplast and apoplast together.

Translocation is the function of entry and retention



of herbicide molecules in the conducting element for sufficient longer time so that these can move away from the site of entry to the other tissues in the plant. There shouldn’t be compartmentalization of the herbicide in the tissue or even in the soil. High proportion of weakly acidic herbicide which are of intermediate permeability retained by the cell can be translocated in the phloem. Herbicides that are not retained in the plant cells are more likely to be translocated in the xylem. Lipophilic molecules may enter the phloem cells more rapidly than more polar molecules, but are also more likely to efflux from phloem soon after their entry. While other molecules may penetrate the phloem element more slowly, but are more likely to be retained and translocated in the phloem. There can’t be herbicide that is completely phloem mobile. Any herbicide that possesses optimum characteristics for phloem mobility is also able to efflux from the phloem and move along the xylem. When herbicide

enters and start to move in the phloem, leakage up to the xylem or surrounding tissue can occur at any time depending on the rate of flow in the phloem and length of the translocated system. In some instance herbicide can be cycled or re-translocated. As some herbicides translocated in the phloem and can efflux to the xylem and may accumulate in the young leaves and can again be exported from those leaves when mature to other system. But it depends on its remaining available for translocation over an extended period of time.

Factors affecting absorption and translocation of foliage applied herbicides are plant structure, plant morphology, relative humidity, spray drift, vaporization and run off/ wash off. Factors affecting absorption and translocation of soil applied herbicides are adsorption, vaporization, photodecomposition, leaching, chemical decomposition, microbial decomposition and soil moisture.

FIG. Different barriers and transformation of herbicides after application

Herbicide MetabolismHerbicide metabolism is the method of transformation rather than detoxification of herbicide inside living system. Herbicide resistance is also defined as “the sum total of a series of simultaneous and/or sequential reactions leading to structural modifications and thereby functional modification of a chemical inside the living system, animals or plants. It may be considered as the basis of selectivity of herbicides inside the plants.

Metabolism/degradation of herbicides in plants is mainly of two types i.e. enzymatic degradation and non-enzymatic or chemical degradation. Enzymes involved in enzymatic degradation i.e. Glycosidase (many herbicides), Glucuronidase (for Carbamates), Glusulase (for Pheols), N-glycosyl transferase (for Benzoic), Glutathione-S-transferase (Triazines), Carboxylesterase (Aryloxyphenoxypropionates), Monooxygenase (thio-, dithio- carbamates and methyl thio triazines), Oxidase etc. An ideal example of non-enzymatic herbicide degradation is Benzoxazinone mediated hydroxylation of triazines detoxification in maize. There are different Degradation reactions

operated in plants enlisted below;

� Oxidation: Phenoxy alkanoic acid herbicides undergo α-, β-, γ- oxidation and get detoxified. Whereas, β-oxidation for compounds like 2,4-DB and MCPB, having even number of C-atoms in the side chain results in increased toxicity to plants called “Reverse metabolism”.

� Hydrolysis: It is operative mainly in the esterified products of herbicides and a common degradation pathway of carbamates, thiocarbamates, triazines, substituted phenyl urease and sulfonyl urease. Ex: Propanil hydrolyzed to non-toxic 3,4-dichloro aniline by arylacylamidase enzyme in rice which is reported 20-30 times greater than in Echinochloa sp. (susceptible).

� N-Dealkylation: N-dealkylation results in the removal/substitution of alkyl groups e.g. methyl, methoxy, ethyl, ethoxy, propyl, iso-propyl etc. attached to the N-position of several herbicides. It usually occurs in carbamates, thiocarbamates, phenyl urease, dinitroanilines and pyridazinones.

� Conjugation: Aggregation/binding up of



herbicides with several parent compounds or their metabolites in plants. Conjugates have higher molecular weight are less mobile and therefore, their translocation to the site of action is retarded which results in differential selectivity. Glutathione-S-transferase catalyzes the conjugation reaction in maize, sugarcane and sorghum which make them tolerant to simazine.

� Ring cleavage: Here the aromatic heterocyclic ring structure is split/broken resulting in the formation of a less phytotoxic compound in plants. It’s a very slow process and doesn’t appear to be major process in herbicide transformation. Triazines and phenoxy alkanoic acids are subjected to ring cleavage.

� De-carboxylation: It is the removal of “-COOH” group or release of CO2 from the chemical or replacement of “-COOH” group by other functional groups. Decarboxylation is operative in phenoxy alkanoic acids and CO2 is released from the herbicide in presence of water.

� Deamination (Metamitron, Metribuzin): It renders the removal of “amino group” (-NH2) from the herbicide for its detoxification. This reaction is

catalyzed by peroxisome-based deaminase.

ConclusionDifferential rate of absorption, translocation and metabolism are the functions of herbicide selectivity. The symplast moving herbicides should recommend at proper time and at optimum rate for its efficient action. There are different herbicide deactivation mechanisms present inside the plant system for their tolerance to phytotoxic effect of different herbicide, among which enzymatic degradation is most important.

ReferencesShimabukuro, R.H. 1985. Detoxification of herbicides,

Weed physiology, II: 216-240.Cellular absorption of herbicides, Plant and soil

sciences e-Library. Accessed on 2nd Dec. 2016.Rana, S. S. and Rana, M. C. 2015. Advances in weed

management. Department of Agronomy. College of Agriculture, CSK Himachal Pradesh Krishi Vishwavidyalaya, Palampur, India.

Das, T.K. 2013. Weed science basics and applications. Jain Brothers publication, Karol Bagh, New Delhi, India.

AGROMETEOROLOGY, REMOTE SENSING & GIS

19706

22. Crop Cutting Experiments (CCE)G. SRINIVASAN1, A. KARTHIKKUMAR2 AND B. SABARINATHAN3

1Ph.D. Scholar (Agronomy), ACRI, TNAU, Coimbatore- 641003 2SRF, RS&GIS, ACRI, TNAU, Coimbatore- 641003 3M.Sc. Scholar, RS&GIS, ACRI, TNAU, Coimbatore- 641003

Crop Area StatisticsThe data on crop area measurements is the foundation of our Agricultural statistics. Dependable and convenient information on crop area is vital to organizers and strategy producers for proficient and opportune agricultural and horticultural improvement and settling on significant choices as for acquisition, safe stockpiling, open circulation, fare, import and other related issues.

IntroductionCrop Cutting Experiments (CCE) is are used in crop insurance to achieve a realistic reliable and precise estimation of crop yield. Which is of particular importance. The Indian government uses this knowledge for its Pradhan Mantri Fasal Bima Yojana (PMFBY) which is a program that flawlessly lets insurance companies dispense installments for farmers insurance claims. The PMFBY to claim insurance each state should have to carry out at least four CCEs for each crop at each village level and send details on yields to insurance companies within a

month of harvest. Such experiments are performed by the agricultural statistics department of each state using stratified random sampling technique each block being taken as a primary unit. Specific yield of a region can be determined based on CCE.

Objective of the schemePMFBY aims for supporting sustainable production in agriculture viz.,

� Providing financial support to farmers for crop loss/damage arising by natural calamities

� Stabilizing the farmer’s income to ensure their continuance in farming

� Encouraging the farmers to adopt innovative and modern agricultural practices

� Ensuring flow of credit to the agriculture sector which will contribute to food security, crop diversification and enhancing growth and competitiveness of agriculture sector besides protecting farmers from production risks.



ProcedureThe CCE or Crop Cutting Experiment is a tool used to evaluate the village’s total yield. Once a village is selected, CCE conducted for all notified crop. They must send one copy of Form II to their offices on harvest day, viz. Tahsildar, BDO, Agricultural officer. CCE should be done in supervision and presence of Village Level Committee (VLC). If the selected village does not have at least two survey number of the chosen (experimental) crop, then “No Crop” report has to be produced by field officers and a request should be made for a for change of village. In the case ofiZero productivity, VLC has to report this as Zero Production”

Data Collected in CCE: - Three informative forms are filled during CCE Experiment.

• Form - Marking of the crop.• Form II - Recording crop yield.• Form III - Harvesting details

Stages of CCE: Selection of village, survey/sub-survey number and plot were done randomly.

Conditions for Conducting CCEVillage should have atleast two survey/sub-survey number growing the notified crop. The selected plots should not be grown for seed purpose/crop competition/exhibition/fodder purpose. If the selected field is a mixed crop, then the crop selected for CCE should be more than 10%. Area of crop grown should be more than 10mx5m or 5mx5m as per the crop selected. CCE must be conducted in all conditions except prevented sowing as per guidelines of PMFBY.

Plot size and method: 10mx5m For Tur/Redgram, Cotton, Sunflower, Caster And Tobacco -5mx5m For All Other CropsThere are two methods of conducting CCE namely, Row method and Non-row method

1) Plot measurement for CCE 2) In-situ weighing of CCE yield

Primary worker allocated for conducting of CCEs: Revenue Village accountant, AAO/AO of Agriculture, HO/AHO of Horticulture and Secretary/PDOiof RDPR departments.

Drawbacks of Traditional Method of CCEParticularly in a scenario where there are nearly 2.5 lakh gram panchayats scattered in India, along with insufficient qualified human labour or time to effectively promote these experiments, there needs to be a more efficient way of use the resources and obtaining an accurate yield estimate within the short harvest period.

Digital Platforms in CCEsNowadays CCE data were submitted to insurance companies through the digital manner by the Crop Cutting Experiment app.

PMFBY (Mobile app)

Risk coverage � Prevented Sowing/ Planting Risk: � Standing Crop (Sowing to Harvesting): � Post-Harvest Losses: � Localized Calamities � General Exclusions

How Technology Makes it EasierThe use of new technology in agriculture has made the practice of farming a lot more predictable and efficient. Remote Sensing and other technological advancements in agriculture provide a far more accurate and timely estimation of yield while compare with use of random sampling in the traditional method of CCE, the impact of technology starts much earlier. products such as like Note cam captures the precise location, farmers details



and crop information right from the pre-harvest stage for conducting CCE experiments. This ensures that the field data is reliable, allowing authorities to use specific data conveniently at the correct time

ConclusionGovernments will use CCE for prepare agricultural policies and services for the future, the information will assist financial institutions with all the resources they need before providing loans or insurance coverage if the harvest or crop failure is low.

WEED SCIENCE

20108

23. Cropping System: A Component in Cultural Method of Weed ManagementVARSHINI S. V.

PhD scholar, Department of agronomy, TNAU Coimbatore. *Corresponding Author Email: [email protected]

It is a known fact that weed management is considered to be a factor in any farming system. The proper strategy for weed control should be altering the competitive balance between the crop and the weeds through measures such as correct choice of rotation, choice of crop species and variety, appropriate sowing arrangements, under sowing and other prophylactic weed management measures (Younie & Litterick, 2002).

Crop RotationCrop rotation in simpler terms is the processes of rotating crops in a same field at a planned sequence year after year. Crop rotation enables crop diversity which in turn increase the sustainability of the system. Crop rotation can either be planned or dynamic in which farmers adapt their crop rotations in response to environmental, pest and market forces. Crop rotation has the potential for enabling the foundation for long-term weed management.

Inter CroppingIntercropping is a method of cultivating more than one crop species simultaneously in the same field. This diversifies production and also increases the net income through proficient consumption of all available resources. Intercropping has a vast advantage over monocropping systems because of their efficient competition for the available resources or to their alellophatic effect on weeds. Intercropping systems further utilize the resources not exploited by weeds or might better convert such resources to the economic part of the crop.

Cover CropsThe term cover crop refers to growing plants as cover in rotation during when the main crops are not grown. These cover crops are usually killed (mechanically or chemically) before going for the main crop. Whereas living mulches are cover crops that are planted between the rows of a main crop and are maintained as a living

ground cover throughout the growing season of the main crop. Although living mulches are considered to be a crop cover they are grown throughout the lifespan of the main crops. Apart from the definitions, both cover crop and living mulch suppress weeds by the similar mechanisms.

Agricultural Practices-Planting Arrangement

Spatial uniformityThe crop spatial uniformity helps in decreasing the competition within the crop population early in the growing seasons and also maximizes the total shade cast by the crop by reducing self-shading.

Planting densityThe practice of increasing crop plant density by using higher seeding rates associated with narrower row spacing can lead to earlier canopy closure, thus shading weeds in their early developmental stages.

Row orientationLight is an important determinant of crop productivity. Crops can be manipulated to increase shading of weeds by the crop canopy, to suppress weed growth, and to maximize crop yield. In general, cropping systems that reduce the quantity and quality of light in the weed canopy zone suppress weed growth and reduce competition.

ConclusionThere are so many strategies that are helpful in controlling numerous weeds on different areas under different conditions. But there is no particular method which can completely eradicate weeds without creating any unintentional outcomes. The underlying basic concept behind weed management is to make the plants dominant than the weeds in the competition for nutrition. Even though the cultural methods of weed management are best the way to control weeds without creating much harmfulness to the plants they are no



way as efficient to meet the modern needs. Practices such as crop rotation, intercropping, cover cropping are not only successful in weed management but also economically beneficial for the farmers. Allelopathy is the next big thing in terms of weed management

which needs a lot of attention and research but shows a promising outcome so far.

ReferencesYounie, D. & Litterick, A. 2002. Crop protection in

organic farming. Pest. Outlook 13(4), 158–1.

ORGANIC FARMING

20092

24. Potentialities of Use of Liquid Manures in AgricultureAVIJIT GHOSH, MANJANAGOUDA S. SANNAGOUDAR, HANAMANT M HALLI AND KEERTHI, M. C.

Scientist, ICAR-Indian Grassland and Fodder Research Institute, Jhansi (U.P.)-284003

IntroductionIndia is an agricultural nation and a big consumer of water resources for irrigation. But there is a strong demand for irrigation of water, while gallons and gallons of effluent are released into water supplies as untreated. Sugar mills, thermal power plants, paper mills, textiles, distilleries, fertilizer systems, electroplating plants, tanning industries, sago factories, oil refineries, pesticides and herbicide industries. The chemical effluents containing heavy metals pose a significant threat to the environment. The use of these industrial effluents and sewage sludge for agriculture has become standard practice in India, as a result of which these toxic metals are transferred and absorbed from soil to plant tissues. In the era of globalization and industrialization, there is an increasing demand for good quality produce and pollution free environment. On the contrary, many of the industries make use of large quantity of good quality water and discharge as waste water, with the objectionable odour and harmful constituents which may pose threat to natural resources. Soil and water are the two important natural resources which hold key role for safe living. Majority of industries are agro-based, demand by these industries for water is expected to increase from 5 % in 2000 to 23 % by 2025. Recycling of industrial and sewage effluent for crop production has dual advantage. In this context, it is worth noting that nutrient management through liquid manures viz. spent wash, paper mill effluent, coffee pulp effluent, sewage, jeevamrutha, and bio-digested liquid manure, etc., play a major role to achieve sustained soil fertility and crop productivity by maintaining soil health by build-up of soil organic matter, beneficial microbes, enzymes, besides improving soil physical and chemical properties. Liquid manures can be effectively and economically used in crop production.

Imperatives of using Liquid Manures � The cost of fertilizer has significantly increased

the market for fertilizer use. � Indiscriminate application of fertilizer has

impaired the health of the soil. � The supply of organic manure is minimal and the

organic carbon content of semi-arid tropical soils is very small.

� The soil shortage of micronutrients is widespread. � Decline in yield of crops � Lack of availability of good quality water for

irrigation � Increased production of highly polluting industrial

wastes, posing disposal problemsThe alternate approach to address the above

problem is scientific use of liquid manures in agriculture.

What is liquid manure?Liquid manure is manure in liquid form. It contains less than 10% of the solids. Manure is converted into a liquid form by combining the manure with the water. Liquid manure is commonly used as a convenient alternative to solid manure, which cannot be distributed equally in contrast to solid manure, which is used as a nutrient-enriched fertilizer for plants.

Different types of liquid manures � Distillery effluent � Sugar mill effluent � Coffee pulp effluent � Paper mill effluent � Anthropogenic liquid waste � Sewage � Jeevamrutha � Bio digester Liquid Manure

Advantages of liquid manures � Helps in improving growth and yield of the crops. � Improves the quality of the produce. � Eco-friendly in nature � Helps in buildup of soil organic carbon



� Wholesome improvement in the soil health

Constraints in using liquid manures � Effluents treatment requires huge cost � Transportation of liquid manures to fields is

problematic � Lack of technical knowledge for famers to use the

liquid manures � Application of industrial effluents needs proper

dilution � Sugar mill effluent is having unacceptable colour

and odour which hinders the acceptance by farmers.

� Coffee pulp effluent and distillery effluent is highly acidic and it is having high BOD and COD levels

� Paper mill effluent contains higher percentage of sodium which affects the soil health and productivity

� Untreated Sewage water contains heavy metals such as Pb, Cd. Which hinders crop growth and development

� Collection and handling of human urine posing social problems

Conclusion � Liquid manures have huge potential to be used

safely in agriculture. � Coffee pulp effluent can be used where soils are

poor in nutrient status and is comparatively less harmful when effluent alternate irrigated with fresh water.

� Application of sugar mill effluent to the agriculture / horticulture filed as a source of irrigation is a viable option for safe disposal of effluent with improvement in soil physical and nutrient status.

� Human urine can be used as liquid manure and it can supplement to fertilizers.

CROP ECOLOGY AND ENVIRONMENT

20213

25. Carbon SequestrationAMRUTLAL R. KHAIRE*, PRASANTAKUMAR MAZHI AND SONALI V. HABDE

Research Scholar, Banaras Hindu University, Varanasi *Corresponding Author Email: [email protected]

Carbon dioxide (CO2) is one of amongst important

greenhouse gases which is essential to maintain the earth’s temperature at normal level. Without it the planet earth would be extremely cold. Although this is fact, but gradual increase in CO

2 is threat for global

warming ultimately disrupt our earth’s climate. According to IPCC 2007, since 90th globe temperature raised by 0.8 c֯ and it is 11th warmest year in world. Anthropogenic activities are highly responsible for the climate change. To tackle with such jeopardous situation there is systematic process called as carbon sequestration.

What is Carbon Sequestration?The scientific method to capture waste CO

2 from its

emission point, transporting it to storage site and unload it, from where it will not return to atmosphere or it can slowly enter to the atmosphere is called as carbon sequestration. Simply it is long term removal and capture of CO

2 so it is also called as Carbon Dioxide

Removal (CDR).

Types of Carbon SequestrationAccording to the methods of storage of CO

2 carbon

sequestration can be divided into following type:a) Geo-sequestration b) Ocean sequestration c)

Mineral sequestration d) Plant sequestration e) Soil

sequestration

1. Geosequestration: It is process of collection and storage of carbon di oxide into suitable underground formation for storage. It is depleted in oil and gas reservoirs, saline formations, or deep, un-minable coal beds. It consists following stages: a) Capturing of CO2: captured from gas or coal power plant. It may be from burning of fossil fuels and other industries like cement, plastic etc. b) Transport and storage: the collected CO2 is converted into supercritical fluid and compressed it at =100bars. So that It will easy for transport. c) Injection: the CO2 is then injected into the ground up to 1KM deep so that it will be for many years

2. Ocean Sequestration: Ocean is considered as largest carbon stored on the earth. Also 1/3rd of CO2 releases in atmosphere directly enters in the ocean. But improper method and extra carbon dissolution make the water acidic and also create the problems flora and fauna in the ocean. In this process either the CO2 dissolution is done in water column at dept of 1000m and CO2 will dissolve subsequently or the CO2 lake will be form by injecting to sea floor at depth greater than 3000m. this will help to slow down the release of CO2 in environment. Still the sequestration in the ocean is require further research in biological as well as chemical views.

3. Mineral Sequestration: The formation of stable



carbonate by reacting CO2 with metals oxides and formation of carbonate mineral which is stable in nature is called as Mineral sequestration or Mineral carbonation. This process is exothermic type and occur naturally. Involved weathering reaction is slow so energy intensive reactor may be used.

4. Plant Sequestration: Carbon plays important role in photosynthesis process in plant. That CO2 involve in this process is stored in the form of biomass called as plant sequestration. This can be enhanced through forestry practices and revegetation. It is significant short-term contribution to carbon sequestration to mitigate problems of climate change.

5. Soil Sequestration: Soil is reach source of carbon. But due to conversion of forest soils into agriculture land there is loss in soil carbon content by more than 40%. This can be counteracted by appropriate adoption of practices. Managing agricultural practices and procedure to increase the organic content in soil is called as soil carbon sequestration.Practices include zero tillage/low tillage, use of

organic manures and compost, avoid stubble burning, crop rotation, cover crop, diverse culture system than monoculture, perennial vegetation on slopes and steep, Bamboo farming, encourage organic and intensive farming. Use of Biochar is also one of the important approach for soil sequestration. This will improve buffer pH, complexes cation action, immobilized pollutants, binds heavy metals. It will provide encouragement for biological process, large storage N, P, K, S, Ca, Cu, Zn etc., improvement soil resilience. This will develop physical qualities structural ability of soil, influence water retention, buffers soil temperature.

What is Biochar?Biochar is granular, solid and stable charcoal like material developed by pyrolysis (incomplete heating) of biomass. It is well adopted option as slash and char rather than slash and burn. One of the approach in soil, include incomplete burning of organic material such as crop residue, wood chips, manure, municipal waste, stubbles etc. in an oxygen limited environment. Biochar is chemically stable in soil for thousands of years. Here biochar helps to convert labile carbon to stable form. The benefits of biochar are addition of nutrient in soil, increase in water holding capacity, increase soil acidity, reduction in use of fertilizer and irrigation etc.

ConclusionThe sink capacity needs to enhanced by forestation and manipulation of biological processes effectively and deliberately towards carbon sequestration is ultimate need for carbon. Governing over anthropogenic activities to less carbon emission is highly required. Integrated systematic situation will be highly effective which concern with geological, ocean, biological sequestration. There is highly requirement to make these technologies which will be cost effective, safe for environment, reduce leakage risk, free from adverse impact. Use of biochar, biodiesel, biofuel etc. are some approaches need to follow for reducing carbon emission.

ReferenceRatan lal, 2008, Carbon sequestration USA Phil.

Trans. R. Soc. B 363, 815–830h t t p s : / / w w w . u s g s . g o v / f a q s / w h a t - c a r b o n -

sequestration?qt-news_science_products=0#qt-news_science_product

CROP PHYSIOLOGY

20136

26. Physiological Role of Chitosan in PlantsBINNY SHARMA AND PAYAL CHAKRABORTY

Research Scholar, Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi *Corresponding Author Email: [email protected]

IntroductionChitosan is a linear polysaccharide composed of β-(1-4)-linked D-glucosamine and N-acetyl-D-glucosamine linked to each other through 1.4-glycosidic bonds. Chitosan is obtained from N-deacetylation of chitin. Chitin is a natural polysaccharide extracted from shells of shrimps, crustaceans and other organisms like insects and fungi. Chitosan is a naturally occurring polymer widely used in medical industries, food industries, cosmetics, biotechnology etc. Biopolymer

chitosan is receiving much intention nowdays for its wide application in agriculture due to its excellent biocompatibility, biodegradability and bioactivity respectively.

FIGURE 1: Structure of chitosan



Physiological Role of Chitosan in PlantsChitosan and its derivatives induces several physiological and biological responses and its effect has been studied in several developmental stages of plant. The role of chitosan has been extensively studied in many crop species like cereals, horticultural, ornamental and medicinal plants. Seed priming with 0.05-0.2% chitosan increased the germination percentage, germination rate and seedling growth in Pimpinella anisum (Mahdavi, 2016). Chitosan oligosaccharides enhances yield components and production quality of wheat (Wang, 2015). In mung bean, the application of chitosan (50ppm) increases photosynthesis, harvest index, chlorophyll content, nitrate reductase activity and yield attributes (Mondal et al., 2013). Thus, chitin and its polymers are widely used to ameliorate crop production and conservation of agronomic commodities.

TABLE 1: Effect of chitosan and its derivatives in plant species

Plant Species Chitosan and its derivatives Effect on plants

Arabidopsis CH Oligomers (tetramers)

Increased fresh weight, radicle length, total carbon and nitrogen content

T. aestivum Nanochitin whisker Yield, grain, protein, iron and Zn content

Soybean Chitoligosaccharides+ Bradyrhizobium

Enhanced nodule formation, dry mass of roots

Ginkgo biloba L. CHT/nano-TiO2 & CHT/ nano SiO2

Invitro preservation of seed quality

Zizyphus jujube Mill cv. Dong Zao

CHT/ nano-silica/ sodium alginate

Extend fruit shelf-life, inhibition of pathogen growth

Role of Chitosan in Abiotic Stress ResponsesChitosan elicits several defense responses in plants related to abiotic and biotic stresses. They not only regulate plant physiological and developmental processes but also stimulates stress tolerance in plants. However, the mechanism whereby chitosan elicits defense responses is not well understood. Chitin-specific receptors are present in plant cell membranes

which are known to elicit defense responses. When treated with chitin-based treatment, plants activate their defense mechanism since they mimic compounds related to chitin-containing organism (Hindangmyum et al., 2019). Table 2 represents the effect on chitosan on different crops in stress responses.

TABLE2: Effect of chitosan on abiotic stress in different crop plants

Plant Species Chitosan Abiotic Stress Effects

Brassica rapa L. 1kDa Cadmium stress

Enhanced antioxidant and photosynthetic activity

Petunia × atkinsiana D. don

970 kDa Salinity Stimulates shoot growth in vitro

Chrysanthemum Robinin + chitosan

Drought Improves plant growth and water stress tolerance

Phaseolus 0.1 and 0.3% nano chitosan

Salt stress Promotes seed germination & radical length

Conclusion: Chitosan an important biopolymer has been reported to stimulate several developmental processes in plants and elicits several defense systems as well during abiotic stresses. Chitosan and its derivatives control various phases of plant life cycle from germination to yield. Chitin-specific receptors are present in plant cell membranes which are known to elicit defense responses but mechanism of of elicition needed to be explored greatly.

ReferencesMahdavi, B (2016). Effects of Priming Treatments

on Germination and Seedling Growth of Anise (Pimpinella anisum L.). Agriculture Science Developments, 5(3), 28-32.

Mondal, M.M.A., Malek, A., Puteh, A & Ismail, M. (2013). Foliar application of chitosan on growth and yield attributes of mungbean (Vigna radiata (L.) Wilczek). Bangladesh Journal of Botany, 42(1), 179-183.

Hindangmyum, A., Dwivedi, P., Katiyar, D & Hemantranjan, A. (2019). Application of chitosan on plant responses with special reference to abiotic stress. Physiology and Molecular Biology of Plants, https://doi.org/10.1007/s12298-018-0633-1.



20190

27. Yellowing of Soybean and its Management from Farmer’s PerspectiveL. S. RAJPUT1*, SANJEEV K2 AND V. NATARAJ3

1,3Scientist (1,2Plant Pathology, and 3Genetics), ICAR-IISR, Indore (MP) *Corresponding Author Email: [email protected]

IntroductionDue to high protein and oil content in soybean, today world has enormous potential to combat malnutrition problem through soybean consumption. Among oilseeds, soybean obtained top position in oilseed production in India, despite its commercialization only since last four decades and acreage mostly limited to five states only. In the changed scenario, a new problem in soybean is being reported by extension workers, scientists, private companies as well as farmers referred as yellowing of soybean. The yellowing in soybean is caused by biotic as well as abiotic factors. The biotic factors include diseases and insect pests; whereas abiotic factors include water logging, drought and nutritional deficiency.

Biotic Factors

DiseasesThe yellowing symptom in soybean occurs mainly due to diseases caused by fungi and virus. The fungal diseases showing such symptoms are anthracnose, rust, fusarium wilt, alternaria leaf spot and charcoal rot, whereas viral disease include soybean yellow mosaic virus disease. Since last three years across the India, anthracnose, rust and soybean yellow mosaic virus diseases have reduced the soybean yield drastically. The Anthracnose disease exhibits symptoms like brown spot surrounded by huge yellowing along with veinal necrosis, whereas soybean rust produces tiny chlorotic spots, greyish brown in colour on both the surface of leaves but more often on lower surface, gradually spots become pustules and produce powdery fungal spore mass while touching. Fusarium wilt can be easily identified as yellowing of leaves with partially wilting of soybean along with rotting in internal tissues of stem. Alternaria leaf spot produces black circular spots in concentric fashion along with yellowing of leaves. Charcoal rot express symptoms under drought condition by slight yellowing of leaves with blacking of stem with huge production of sclerotia on stem and roots. These microsclerotia can be easily observed when the stem is split open. These diseases can be managed by seed treatment with either thiophanate methyl+pyraclostrobin @ 2 ml or carboxin+thiram @ 3 g/kg of soybean seeds, followed by spraying of tebuconazole @ 625 mL/ha or tebuconazole + sulphur @ 1000g/ha hexaconazole @ 800 mL/ha or pyraclostrobin @ 500 g/ha at 45 and 60 days after sowing (DAS). Soybean yellow mosaic

virus disease causes yellowing of leaves alternating with green patches which turn complete yellow at later stage. YMVD can be managed by seed treatment with imidacloprid 48 FS @ 1.25 mL/kg or thiamethoxam 30 FS @ 10 mL/kg of soybean seed followed by spray of thiamethoxam 25WG @ 100 g/ha (Rajput et al., 2019).

Insect PestsSoybean whitefly and aphids feed on plant juice through sucking; simultaneously they transmit virus and inject toxins in the soybean plants. The transmitted virus and toxin produce yellowing symptoms on leaf surface. This can be easily identified by insect pest occurrence near the soybean plant. Soybean whitefly and aphids can be managed by seed treatment with imidacloprid 48 FS @ 1.25 mL/kg or thiamethoxam 30 FS @ 10 mL/kg of soybean seed followed by spray of thiamethoxam 25WG @ 100 g/ha. A slight yellowing with wilting is observed during white grub infestation. This can be identified by easy uprooting of soybean plant and absence of lateral roots. The white grub can be managed through integrated pest management only viz, avoiding use of unripe FYM, seed treatment with imidacloprid 48 FS @ 1.25 mL/kg followed by installing of light trap for destruction of insect and soil application of chlorpyriphos @ 16 kg/ha after 25 to 30 DAS.

Abiotic Factors

Waterlogging and droughtThe waterlogging in soybean is a complex phenomenon that includes continuous raining in soybean field. The waterlogging can occur even in plain field due to non-leveling of field. Such waterlogging condition results in yellowing of leaves along with wilting and drooping of erect soybean plants. It can happen either in single plant or randomly in small patches in the field. Such plants showing aerial root initiation on stem due to low oxygen availability in soil where initially small protuberance can be observed in stem nearly 10-15 days after heavy rain. Sometime farmers also face shortage of water due to uneven distribution of monsoon accompanied with high temperature. Such condition also results in wilting of plant along with slight yellowing of leaves. Such waterlogging and drought condition can be managed by leveling of field, growing of soybean on broad bed furrow or ridge and furrow system. The application of subsoiler once in three year can reduce the adverse effect of both waterlogging and



drought. At the time of waterlogging, application of nutrient induces growth in plant, alongwith providing drainage facility.

Nutritional deficiencyMostly in early stage of crop growth calcium induces iron deficiency leading to complete yellowing of young leaves (10-20 days old seeding) while veins remain

green during drought condition. It can be managed by application of foliar spray of FeSO

4 @ 0.1 % with

irrigation.

ReferenceRajput L.S., Sanjeev K, Meena L.K., Raghavendra

M, Verma R.K. and V. Natraj 2019, Chemical management of biotic stress in soybean, Agrobios Newsletter, 18 (7): pp 17-18.

20191

28. Plant Phenomics: An Emerging Contemporary Approach to Combat Abiotic Stress in CropsAKANKHYA GURU1*, SOUMYA KUMAR SAHOO2, AND SELUKASH PARIDA3

1Department of Plant Physiology, IAS, Banaras Hindu University, Varanasi 2Department of Plant Physiology, COA, Indira Gandhi Krishi Vishwavidyalaya, Raipur 3Department of Plant Physiology, COA, Odisha University of Agriculture & Technology, Bhubaneswar *Corresponding Author Email: [email protected]

IntroductionBy 2050, according to an estimation, human population will reach 9 billion. To meet the basic requirements of the growing population, current food production must be doubled up. In the present context, the harmful impacts of climate change have aggravated the challenge posed by environmental stresses related to global food production. Despite adverse environmental conditions and a limited area of cultivation, the worldwide challenge to feed the growing human population demands a consistent increase in crop production. Yield potential and stability of a genotype are central characteristics for increasing the production of crops. Abiotic stress tolerance plays an imperative role in stabilizing plant performance. The impact of various abiotic stresses on different crop plants can be studied by performing experiments either in field or under controlled environments. Plant phenotypic responses under stressed conditions must be examined to determine the potential of a particular genotype. Phenotypic changes in plants can be measured by examining the key parameters such as root morphology, leaf related traits, yield associated traits etc. that are stress-specific, through the employment of sophisticated techniques and hence resulting in a precise estimation of the plant phenotypic response. Errors in measurements may lead to unauthentic ranking of the examined genotypes. In order to achieve a breakthrough success in crop improvement, novel phenotyping tools are required that can record measure phenotypic changes occurring in plants under stress.

Before switching to plant phenomics, we need to understand what is a plant phenome?

Plant Phenome – It is defined as the plant traits that are resulted from the gained knowledge of

the genetic program stored in the cell under the given environmental conditions. For example, dimension, shape, position, cell/organ/plants orientation, biochemical characteristics etc.

Plant PhenomicsThe scientific study of the characterization of plant phenome is known as ‘plant phenomics’. It involves various disciplines such as biology, physics, genetics, computer science, statistics, meteorology as well as other related disciplines. Plant phenomics consists of five different pillars:1. High throughput data acquisition – It is

required for cells, organs, individual plants, and plant canopy including both the root system and the aerial parts of all plant species, under controlled or field conditions.

2. Data management – ontology, meta-information, information sharing

3. Data interpretation – It includes machine learning, statistics, computer vision, signal processing, data fusion, scaling

4. Modeling – meta-analysis, sensitivity analysis, data assimilation, dynamics of plant structure, eco-physiological processes

5. Applications – plant breeding, resource management, precision agriculture, decision support

Imaging Techniques involved in Plant Phenomics � Visible light imaging – The wavelength of

visible light ranges from 300-700 nm. It offers advantages such as cost-effective, easy to handle. It relies on both two-dimensional and three-dimensional digital images. Two-dimensional imaging measures shoot, root and yield related



traits whereas three-dimensional system measures leaf morphology, shoot dry weight, plant height. Various phenotyping platforms have been used for different abiotic stresses in plants:

TABLE 1. Platforms for plant phenotyping in response to abiotic stresses in different crops

Platform Crop Abiotic stressPHENOPSIS, WIWAM Arabidopsis DroughtLemna Tec Barley, maize DroughtLemna Tec Rice, barley, wheat SalinityPlantScreen Arabidopsis ChillingGROWSCREEN Peas Cold injury

� Infrared and thermal based imaging – This technology uses thermal cameras with high thermal sensitivity for detection of canopy temperature in plants. It is used to detect plant pigments, leaf water content, lignin and cellulose. In addition, it is used to study stomatal behavior under salinity and drought stress by observing differences in canopy temperature.

� Fluorescence imaging - Blue wavelength light (< 500 nm) is flashed on the plants and they emit fluorescence light at 600–750nm in the red region of the spectrum. It can be used to study chlorophyll content, stomatal movement, phloem loading and unloading, plant metabolite content. This method can detect stress at the primary level and resolve heterogeneity in photosynthetic performance of leaves.

� Spectroscopy imaging – Using this imaging technique, images are divided into bands and a large fraction of electromagnetic spectrum is created in the images. It is used to measure traits such as water status, pigment content, photosynthetically active biomass in different crop species. It has been effectively used for measuring leaf growth and panicle emergence in rice.

� Integrated imaging –– PET – Positron emission tomography is

used to evaluate photosynthetic performance under stress, biochemical pathways, ion assimilation and transport.

– MRI – Magnetic resonance imaging can be used to capture plant root architecture in pots, internal physiological processes in vivo. It can also measure water diffusion water diffusion and transportation through the xylem and phloem in crops such as tobacco, tomatoes, poplars, and castor beans.

– FRET – Forster resonance energy transfer is another outstanding and advanced technique that is used to detect zinc and calcium dynamics in roots during transportation of sugar.

– PlantEye – It is a high-resolution 3D laser scanner used for scanning plants and creating data related to various traits such as 3-D leaf area, plant height, leaf number.

How phenomics contribute to improvement in plants abiotic stress tolerance?

� Infrared thermography imaging has been used to detect leaf temperature, air temperature and canopy temperature differences under drought and heat stress in tomatoes, melons and lettuce.

� Plant responses to osmotic stress in wheat, salt stress in barley, water stress in rice have been evaluated by using infrared imaging. It has been also used to measure leaf temperature and rate of transpiration in Beta vulgaris under drought stress.

� Leaf spectrometer has been used to monitor photosynthetic efficiency in Nicotiana sylvestris

� Chlorophyll fluorescence and 2D digital imaging have been used to investigate plant responses under drought and chilling stress in Arabidopsis thaliana plants.

Future Challenges and Prospects1. It is needed to focus on complete information

regarding phenomics as information related to the individual phenotype cannot satisfy the analysis of characteristics in the era of ‘omics’.

2. Image-based phenotyping should be advanced by introducing novel tools and techniques based on artificial intelligence. Interpretation, precise evaluation and proper understanding of digital features resulted from an automated phenotyping platform/system are major hindrances in plant phenotyping development as well as application.

3. With a rapid pace and high resolution, we need to study correlations among function of genes, environmental responses and plant performance.

4. Next generation phenotyping platforms will help researchers to gain a deeper knowledge on plants responses under stress conditions.

5. Plant phenomics technologies have an enormous potential to develop enhanced climate resilient crop genotypes in future.

ReferencesSingh, B., Mishra, S., Bohra, A., Joshi, R., & Siddique,

K. H. (2018). Crop phenomics for abiotic stress tolerance in crop plants. In Biochemical, physiological and molecular avenues for combating abiotic Stress tolerance in plants (pp. 277-296). Academic Press.

Ninomiya, S., Baret, F., & Cheng, Z. M. M. (2019). Plant phenomics: emerging transdisciplinary science.

Zhang, Y., Zhao, C., Du, J., Guo, X., Wen, W., Gu, S., & Fan, J. (2019). Crop phenomics: current status and perspectives. Frontiers in Plant Science, 10, 714.



20217

29. Salinity: Key Hurdle in Crop ProductionMEGHA

Ph.D. Scholar, Department of Plant Physiology, G.B.P.U.A.T. Pantnagar, Udhamsinghnagar, Uttrakhand, 263145 *Corresponding Author Email: [email protected]

IntroductionSalinity is one of the most devastating abiotic stresses after the drought. About 6 per cent of land worldwide which is about 800 million hectares is affected by salinity. According to the food and agricultural organization soil is called as saline when electrical conductivity of soil is exceeded above 4ds m-1. In India, about 16 states are effected by either salinity or sodicity in which major saline affected regions includes Gujarat, Haryana, Rajasthan and Maharashtra. Most of the saline regions contain salt naturally in the soil but human-induced activities such as improper agricultural practices, using poor quality water for irrigation and excessive use of chemical fertilizer may accelerate the salt Content in the soil. Predominant salt associated with salinity is Na+ and Cl-. The high concentration of salt in the soil disrupts the uptake mechanism of water and mineral from the soil. Most of the food crops such as rice, wheat, maize, tomato is sensitive to salinity stress. Sensitivity toward the salinity depends on growth stage and genotype of the plant.

Effect of Salinity � Excessive toxicity of Na+ and Cl- � Higher uptake of Na+ and Cl- � Physiological drought � Nutritional imbalance � Reduced osmotic potential � Destruction cell organelle and metabolism

Responses of Salinity

Morphological responses � White leaf tip � Leaf burning, � Leaf browning � chlorosis � Low and late tillering � Spikelet sterility � Delayed flowering � The decrease in leaf area index � Patchy growth � Poor root growth � Less grain weights.

Biochemical responses � Reduced water potential and increased

accumulation of osmolytes such as Proline, glycine betaine, sugars.

� The activity of antioxidant enzymes such as Superoxide dismutase, catalase, peroxidase, glutathione increased.

� Hormone like ABA, Jasmonic acid, ethylene activated.

� Expression of stress-responsive gene, which regulates the uptake of salts.

Physiological responses � Reduced relative water content in leaves � Stomatal closure � Inhibition of germination � Inhibition of Photosynthetic processes � Altered nitrogen metabolism.

Regulatory Mechanism in Plants During Salinity: Plants have following regulatory mechanism to combat the effect of salinity.

� Synthesis of Compatible Solutes: Exposure to salt stress results in accumulation of nitrogen-containing compounds: Amino acids, Polyamines, amides and Carbohydrates such as Glucose, fructose, and sucrose. Major functions of these osmolytes are cellular osmotic adjustment, detoxification of reactive oxygen species, protection of membrane stability and stabilizations of enzymes and proteins.

� Selective accumulation and Exclusion of Ions: An increased level of salt may accumulate in the apoplast of the cell and dehydrates the cell and in cytoplasm restricts enzymes activity which ultimately affects photosynthetic processes. The high concentration of salt is harmful to both Glycophytes and halophytes. Glycophytes limit the uptake of sodium by ion exclusion while halophyte accumulates salt as a mechanism of ion accumulation in vacuoles. Transport of ions from in and out of cytoplasm regulates ion homeostasis. Transport of ion depends on the electrochemical gradient of the cell. Na+ enters tin the vacuole through Na+/H+ Antiporter. Two types of H+ pumps are present in the membrane. SOS1 and SOS2 are salts overly sensitive signalling pathway have an important role in ion homeostasis.

� Induction of Antioxidant Enzymes under Salt Stress: Induction of antioxidants enzymes such as catalase, SOD, GR, APX observed to be increased under salinity. These antioxidant activities vary between genotypes, level of salinity, growth stage and environmental condition also.

� Induction of plant hormone during salinity:



Increased salinity is associated with a decrease in auxin, cytokinin and gibberellic acids while increase activity of ABA and ethylene is reported in increased under salt concentration. ABA causes an alteration in stress-responsive gene which play an important role in the salt-tolerant mechanism.CONCLUSION: Salinity, one of the major

abiotic stresses which affect almost every aspect of physiology and biochemistry of plant through disturbing growth and developmental mechanism of plants. Most of the food crops are sensitive to salinity which ultimately causes a reduction in grain yield. To encounter salinity Stress plants are equipped with various resistance mechanisms but these resistance mechanisms are not enough when salinity levels are severe. Many crops are still sensitive. Various strategies such as soil management, exogenous application of phytohormones, such as brassinolide,

ABA, GA, jasmonic acid and salicylic acid have been used to mitigate the NaCl toxicity but these strategies are not sufficient. Stress inducible promoter needs to be tested for their suitable expression before employs in crop improvement. Due to genetic engineering tool researches are enabling to know the exact pathway and regulatory genes responsible for salt tolerance. As a result, genetic engineering approach had been employed for salinity tolerance.

References1. Moradi, F. and Ismail, A.M. (2007). Responses of

photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice. Annals of botany. 99(6), 1161–1173

2. Yadav, S. et al., (2011). Causes of salinity and plant manifestations to salt stress: a review. Journal of Environmental Biology, 32(5), 667.

CLIMATE CHANGE

20158

30. Technologies for Carbon Sequestration in Agricultural EcosystemHEMANT SAINI* AND POONAM SAINI

Department of Horticulture, CCS Haryana Agricultural University, Hisar. *Corresponding Author Email: [email protected]

In general, carbon sequestration is carbon capturing and the long-term storage of atmospheric carbon dioxide (CO₂) and may refer specifically to:

� A form of geoengineering in which carbon is removed from the atmosphere and deposited in a reservoir.

� Capture and storage of carbon, where carbon dioxide is removed from flue gases before being stored in underground reservoirs (e.g., at power stations).

� Chemical weathering of rocks- natural biogeochemical cycling of carbon between the atmosphere and reservoirs.In petroleum refining or from flue gases from

power generation carbon dioxide may be captured as a pure by-product. CO

2 sequestration includes

the storage part of captured carbon, which refers to large-scale artificial capturing and sequestration of industrially produced CO

2 using subsurface saline

aquifers, ocean water, reservoirs, aging oil fields, or other carbon sinks. Carbon sequestration can be described as long-term storage of CO₂ or other forms of carbon to avoid dangerous climate change and either mitigate or defer global warming. The atmospheric and marine accumulation of greenhouse gases released by burning fossil fuels can be slowed down by carbon sequestration. Though, carbon dioxide is naturally

captured from the atmosphere through biological, chemical or physical processes. Some artificial sequestration techniques and artificial processes are required to exploit these natural processes. These technologies can be grouped into two broad categories: abiotic and biotic sequestration.

(A) Abiotic SequestrationWithout the intervention of living organisms (e.g. plants, microbes) abiotic sequestration is based on physical and chemical reactions and engineering techniques. Having a larger sink capacity than biotic sequestration abiotic strategy of carbon sequestration has received considerable attention. Advance technologies are being developed for CO₂ capture, transport and injection.

(B) Biotic SequestrationThe managed intervention of higher plants and micro-organisms in removing CO₂ from the atmosphere is known as biotic sequestration. It differs from management options which reduce emission or offset emission. Another option for managing the terrestrial carbon pool is increasing use efficiency of resources (e.g. water, energy).



FIGURE 1: A wide range of processes and technological options for carbon sequestration in agricultural, industrial and natural

TABLE 1: Terrestrial carbon management options

Management of terrestrial carbon pool

Sequestration of carbon in terrestrial pool

Reduce the emissions Sequestering emissions as SOC

Eliminate ploughing Increase humification efficiency

Conserve water and decrease irrigation need by integrated pest management to minimize the use of pesticides

Improve soil aggregation

Biological nitrogen fixation to reduce fertilizer use offsetting emissions

Deep incorporation of SOC through establishing deep rooted plants, promoting bioturbation and transfer of DOC into the ground water

Establish biofuel plantations Sequestering emissions as SIC

Bio-digestion to produce CH4 gas

Form secondary carbonates through biogenic processes

Bio-diesel and bioethanol production enhancing use efficiency

Leaching of bicarbonates into the ground water

Precision farmingFertilizer placement and formulationsDrip, sub-irrigation or furrow irrigation

ObstructionDanger of leaksCarbon dioxide may be stored deep underground because at depth, hydrostatic pressure keeps it in a liquid state. The stored gas may act to release into the ocean or atmosphere due to reservoir design faults, rock fissures and tectonic processes.

Financial costsAccording to Intergovernmental Panel on Climate Change the use carbon sequestration would add an additional 1–5 cents of cost per kilowatt hour. If the CCS technology were to be required by regulation, the financial costs of modern coal technology would nearly double. The cost of CCS technology differs with the different types of capture technologies being used and with the different sites that it is implemented in, but the costs tend to increase with CCS capture implementation. With adoption of new technologies these costs could be lowered but would remain slightly higher than prices without CCS technologies.

Energy requirementsThe sequestration processes may consume 25 percent of the plant’s rated 600 megawatt output capacity. The capacity of the coal-fired power plant may be reduced to 457 MW after adding CO

2 capture and compression.

20210

31. Impact of Climate Change on Insect PopulationM. THIYAGARAJAN AND J. KOUSIKA*

*Post-Doctoral Fellow, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu

Insects are cold- blooded organisms which is the important factor deciding their behaviour, distribution, development, reproduction and survival. It is estimated that 200C increase in temperature increases its life cycle from one to five per seasons.

IPCC (2013) has anticipated that the global warming may promote the pest population growth, outbreak frequency and increase in area of infestation which results in greater loss and reduction in global food security. Temperature can cause impact in insect



physiology and development directly or indirectly. Some insects have longer developmental period e.g. cicadas. Some insects tend to complete its life cycle faster during suitable temperature e.g. cabbage moth, onion maggot and European corn borer resulting in more generation and damage. Recently, the Colorado potato beetle has spreaded to northward in Europe whereas its increase in population density was also observed in core European regions. The eastern spruce budworm has moved to north US and reduced damage by nymphs was seen in the south during summer.

For instance, the southern. The green stink bug and spotted stem borer are green stink bug (Nezara viridula) and spotted stem borer replacing the native bugs and borers because of their expansion in south. Similarly, western corn rootworm has expanded till European range and cause heavy ecological damage as they are vectors of maize chlorotic mottle virus, which can spread to other natural hosts.

In case of migratory insects, they may arrive early or extend their overwintering which will in turn cause changes in the behaviour of natural enemies. Reduction in parasitism may occur if the host completes its lifecycle before or non-availability of vulnerable stages for parasitism. In pests like thrips, changes in gender ratio may be prevalent. Insect living in soil may be less affected due to temperature change as they act as insulating medium. Decrease in winter mortality will be found due to warmer winter. Some crops may move to the north as the temperature increases and insects finds expanded host area which in turn increases its diversity. The fossil study has recorded that diversity of insect species and the intensity of their feeding is directly proportional to the increase in temperature. On the other way some insect is more specific to the hosts. Increase in temperature may prevent the farmer to grow the host crop which decreases the pest population. The increase in temperature has same effect on the natural enemy population and has increased attack on insect population. For eg. increase in temperature decreases the response to alarm pheromone in aphid which increases the predation.

The precipitation also decreases the insect population to some extent. Heavy rains wash way insects like onion thrips, water staging will decrease the population of BPH and GLH and drought will decrease the population of pea aphid. Increase in

CO2 increased the insect incidence. Increase in sugar

content of the leaves and decreased nitrogen content will increases the pest damage. Increased CN ratio in leaf tissue decreased the development of insects and increases the chances of parasitism.

Increase in temperature and CO2 in atmosphere increases pest incidence by increasing their horst range, area, lifecycle, damage and reduced overwintering. This will increase the application of insecticides and which increases the chances of resistance development, resurgence and residue problem.

Farmers should keep in mind that climate change is gradual and this will some time to adapt. Farmers who make best use of field monitoring, pest forecasting, recording the pest attack and choosing economically and environmentally friendly control measures will likely to be more successful in managing the pest problem with the changing climate. Entomologist predict that more insect damage will be predicted in temperate regions because of increase in temperature thereby insecticide application will be more in order to bring the population below Economic Threshold Level (ETL) which leads to development of resistance in insects for the insecticides with same mode of action. Hence insecticides with similar mode of action should be used less frequently. The cultural practice used by the farmer for the control of insect pest management may get affected due to change in insect development. The practise of crop rotation may be less effective due to early arrival of the insect or increase in overwintering. Row cover crop should be harvested early as the excessive temperature may damage the crop. However, the effects of increasing temperature will not be uniformly increasing the pest species unless provided with suitable environmental conditions, phenology and life history. This also leads to the decline or extinct of some insect population.

ReferenceIPCC (Intergovernmental Panel on Climate Change).

2013. Working Group I Contribution to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge, UK: Cambridge University Press.



BIODIVERSITY

20171

32. Importance of Traditional Knowledge in Maintaining BiodiversityDEEPIKA PANDEY

Ph.D. Research Scholar, Department of Family Resource Management, GBPUAT, Pantnagar, Uttarakhand

Traditional knowledge is the unique local knowledge, innovations and practices of indigenous communities’ in particular geographical areas around the world. It is being developed through the experiences gained over the centuries and is reflected in the lifestyle and local cultural environment. It is being passed on from generation to generation in the form of cultural values, rituals & customs, laws & beliefs, agricultural practices. The traditional knowledge is basically practical in nature in field of agriculture, forestry, fisheries, horticulture, health and environment. It is important not only to the ones who are dependent on it on daily basis but also to modern industry and agriculture (for example plant-based medicines, health and cosmetics products, handicrafts etc.). Traditional knowledge can be valuable to deal with the causes and consequences of climatic variations and ultimately conserving biodiversity. The advancement in technologies and increasing population has continuously put a strain on the environment and natural resources. Over-exploitation of the resources including water, fuel, land etc. results in their degradation majorly due to deforestation, urbanization, industrial pollution and soil erosion. Hence, it is important to conserve natural resources and environment.

Biodiversity conservation methods can be adopted from various local and indigenous communities where majority of world’s genetic resources are found. Many of these communities have been cultivating the resources in a sustainable way from years ago. Since they have been in direct contact with the biodiversity from years, it created deep understanding of various aspects of biodiversity among them due to which they have been making wise use of natural resources. This has resulted in practicing of various cultural, traditional or spiritual beliefs. Some of the indigenous practices have been found to enhance and conserve biodiversity at local level thereby maintaining healthy ecosystems. Women’s understanding of local biodiversity tends to be broad, containing many unique insights into local species and ecosystems. Women are involved in all the activities of farming starting from seed selection to harvesting to processing and cooking. They are being considered as the managers of natural resources and plays mediator’s role in passing on traditional knowledge. Since women are associated with these aspects and have been in direct contact with natural resources like fuel, water, food and fodder; the contribution of women is very significant in biodiversity conservation.

SOIL SCIENCE

20104

33. Decomposition of Root Residues and Nutrient ReleaseBISWABARA SAHU, ARNAB KUNDU AND SIDDHARTHA MUKHERJEE

Department of Agricultural Chemistry and Soil Science, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia

IntroductionIn terrestrial vegetations, about half of carbon fixed or the photosynthates fixed annually is allocated to belowground pools in the form of root biomass and root exudates. The source of organic matter (either shoot, root or root exudates) along with the quality and

quantity of added biomass regulates the fate of carbon in soil system viz; mineralisation to form CO

2 or to get

stabilised in the form of stable Soil Organic Matter (SOM). But these also affect the fate of metabolic and structural nutrients. The factors affecting the complex phenomenon of organic matter decomposition can



be categorised into three main interacting groups of factors: chemical (composition of the litter), physical (environment and climate of surrounding the litter) and biotic (microorganisms and invertebrates that take part in litter decomposition).

SOM acts as a source of N, P, S and Micronutrients to the soil and plant system. The organic matter added to soil is a rich source of all the nutrients because due to breakdown of the structural and metabolic components of added organic matter into simplified molecules like carbohydrates, amino acids, polypeptides (which undergo further simplification or make stable particles making complexes with other molecules) or in elemental form e.g. K+, Ca+2, Na+, essential micronutrients, etc. Thus, breakdown of these organic matter will directly release the cell bound potassium which is present in ionic form (K+) to the soil solution. Thus, the pattern of decomposition on below ground plant parts can provide clear information towards release of each nutrient.

Factors affecting Root DecompositionThere are few established principles in root decomposition studies but estimating the rate of root decay is bit difficult. To describe the root mass loss through decomposition over time, both exponential and linear decay models have been used. The first stage of root decomposition causes, rapid mass loss. The decomposition of such components depends on inorganic chemical composition of the added biomass and microbial decomposition of water-soluble C for body build up and also some amount of loss through leaching. The second stage is characterized by slower mass loss regulated largely by lignin and other recalcitrant root materials. Unlike leaf litter decay, root decay is less affected by the changing environment effect as it is buffered within the soil system. The soil conditions such as moisture, O

2 concentrations,

pH and direct inorganic nutrient limitations to the decomposer organisms affect the decomposition process though. The chemical and physical parameters like root quality, size of roots affect the decomposition of underground biomass. Root litter quality indices include mineral nutrient concentrations, the concentrations of secondary compounds, as well as C: nutrient and nutrient: nutrient ratios but root quality also differs depending on root diameter. Fine roots are ephemerals and decompose rapidly, but nutrients in structural roots may be released slowly and, thus, may contribute to the growth of new trees in the long term.

Root decomposition and release of nutrientsWhenever there is decomposition of organic matter, the structural components are released very easily. An experiment conducted by Weatherall et al. (2006) showed that 12% of the applied K is moved to a plant of second season from the first season crop residue (to which fertiliser was applied). For P around 70-80% of plant residue P is observed to be released within 1year time of which most part is released within initial 6months period, but this release % is around 90% at the end of 2years. Through such tracer technique he

could find that nutrients released from decomposing roots can make a direct contribution to the growth of restock trees. The rate of decomposition of litter material is dependent on nature of the litter added. It is found that in comparison to leaf litter, roots have a higher concentration of lignin, polyphenols and acid insoluble fractions, but low in P, K and Ca content than leaves. In the initial phases of decomposition, Potassium is released rapidly in both roots and leaves and remains constant thereafter. A study conducted by Fujii and Takeda (2010) showed the pattern of nutrient release as K > Mg > P > Ca = N. As K is not a structural component of cells, its release is less dependent on microbial decomposition. But there is no evidence of K being immobilised in any crop residue. The rapid mobility of potassium may be attributed to lack of incorporation of this element into organic structures. There is evidence of K release at a rate of 92–99% in green manure and 65–95% in other residues. The release of K and P are usually slower in roots as compared to leaves which may be attributed to lower initial concentration on such nutrients in roots as compared to leaves. The position of nutrient in the root may be another factor. E.g. high amount of nitrogen is present in parenchyma cells distributed in cortex which is enclosed with exodermis thus the release is slowed down. As mentioned earlier, high amount of acid insoluble substances is there in roots and Suberin included in such component is recalcitrant which is present in bark and roots. The structure of roots coated with suberin may protect inner plant tissue from microbe attack and thus delay decomposition.

Effect of climatic factors on root decompositionAccording to a study conducted by Silver and Miya (2001), individual climatic and environmental variables are found to significantly influence rates of root decomposition. Mean Annual Temperature (MAT) is positively correlated with root decay rates globally and the relationship was strongest when only fine roots are considered whereas between Mean Annul Precipitation (MAP) or Actual Evapotranspiration (AET) and root decay for fine roots, the relationships are weak and positive one but significant relationship when all root size classes are considered. 25% variability in relationship between AET and root decay for roots of all size class is seen. Both temperature and precipitation were negatively correlated with latitude. Thus, increasing rate of root decomposition is found with decreasing latitude. These factors also influence root decay differently depending on the nature of the plants. E.g. weak positive correlation between root decay of broadleaf species with MAT and MAP for all categories examined. Graminoid root decay has positive correlation with MAT, MAP and AET. While correlating effect of climate on root decomposition and there by cause nutrient release, it is usually observed that K release rate is intensively conditioned by the precipitation during the decomposition period which is found to be higher in rainy months.



ConclusionRoot being a good source of recalcitrant carbon and nutrients, its decomposition has a major role in nutrient cycling, availability to plants and global soil C sequestration. Root chemical, physical structure as well as the climatic factors have specific role for affecting the biogeochemical cycling of root biomass. Thus, Fine root decomposition is mainly characterized by fine root structure which determines the root substrate quality. though release rate is slow but, net N release is much higher in the fine roots. Likewise other nutrients are released faster from fine roots. Still more studies are required to be done to find out actual mechanistic and microbial process level pattern of root decomposition and nutrient release.

ReferencesFujii, S. and Takeda, H. (2010). Dominant effects of

litter substrate quality on the difference between leaf and root decomposition process above-and belowground. Soil Biology and Biochemistry, 42(12): 2224-2230.

Silver, W.L. and Miya, R.K. (2001). Global patterns in root decomposition: comparison of climatic and litter quality TEMPeffects. Oecologia, 129: 407-419.

Weatherall, A., Proe, M. F., Craig, J., Cameron, A. D., McKay, H. M. and Midwood, A. J. (2006). Tracing N, K, Mg and Ca released from decomposing biomass to new tree growth. Part II: a model system simulating root decomposition on clear fell sites. Biomass and Bioenergy, 30(12): 1060-1066.

20155

34. Soil Health Card: A Nuclear Mission Towards Sustainability in AgricultureSUDIP SENGUPTA* AND PARIJAT BHATTACHARYA

Ph.D. Research Scholar, Department of Agricultural Chemistry and Soil Science, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur-741252, Nadia, West Bengal. *Corresponding Author Email: [email protected]

IntroductionSince the green revolution period, injudicious application of high analysis NPK chemical fertilizer with little or no application of organic manure, secondary and micronutrients has led to crop nutritional imbalances and consequently declination in yield, quality and fertilizer response. A significant knowledge gap exists between laboratory and field especially in developing nations like India wherein small and marginal farmers constitute the lion’s share of the community. Accurate determination of nutrient content along with other important contributing factors (i.e. pH, OC) is the pre-requisite for maintaining the soil fertility level as well as conserving the soil health which ultimately comes up with the blueprint of sustainable crop production. Under the Soil Health Management (SHM) programme of National Mission for Sustainable Agriculture (NMSA), the Government of India launched the Soil Health Card (SHC) scheme which is approved by the Department of Agriculture & Co-operation under the Ministry of Agriculture and Farmers’ Welfare.

Soil Health CardSoil Health Card (SHC) is a printed report card available to the farmers for each of his land holdings. It consists of twelve (12) accepted soil testing parameters i.e. macro-nutrients as nitrogen (N), phosphorus (P), and potassium (K); sulphur (S) as the sole secondary-nutrient; micronutrients as zinc (Zn), iron (Fe), copper (Cu), manganese (Mn) and boron (B) and physicochemical parameters as pH, electrical

conductivity (EC) and oxidizable organic carbon (OC).SHC acts as an advisory to the concerned farmers

about the metered application of fertilizers so as for obtaining optimal crop yield. It aims to a) provide a basis to counteract nutrient deficiencies, b) strengthen soil testing network throughout the country, c) identify soil-related constraints, and d) provide economically feasible and ecologically sound nutrient management techniques.

Benefits of the soil health card schemeThe scheme with its manifold advantages benefits the entire food production sector. SHC scheme assists the farmers with scientific future planning for suitable cropping systems based on nutritional demand due to comprehensive monitoring of soil parameters. It monitors the possible changes in the nature of the soil via triennial evaluation. Farmers get to have expert assistance for necessary corrective measures. The precise lacuna of optimal yield and quality from a nutritional point of view can thus easily be identified. It discourages the unnecessary fertilizer application and thereby manifests a twofold gain by minimizing the cost of production as well as the preservation of soil health from chemical abuse.

Sampling, analysis, report generation, and distributionFor analysis of the soil parameters and preparation of SHC, soil samples are collected by trained staff of the state department of agriculture, SAU’s, KVKs, etc in a grid of 2.5 ha in irrigated area and 10 ha in rainfed area



using GPS tools. The samples are generally prescribed to be collected two times in a year, after Rabi and Kharif Crop harvest with no standing crop. Samples are collected with standard “V” shaped cut from a

depth of 15-20 cm, bagged, marked, and sent to soil testing laboratories (STLs) for analysis using standard methodologies for generation of soil health card.

FIGURE 1: Soil Health Card Format

Present ScenarioSHC card scheme, since its beginning (19 February 2015) served through cycle 1 & 2 and model village programme. As on 11th May, 2020; a total of 5,509,973 numbers of samples are analyzed, 49,203,750 numbers of test results are generated and 200,477,306 farmers are taken under. 154,863,559 number of SHCs are entered for the farmer’s utility which serves 66.06% of the target set (234,442,801) (https://soilhealth.dac.gov.in/ Home/ProgressReport).

ConstraintsAlthough the soil health card scheme greatly benefits the farming community, there are some grey areas which should be taken care of. The most important constraints faced by the small and marginal farmers are lack of knowledge and awareness about the use of SHCs, longer time gap between sample collection and report generation, difficulties in fertilizer dosage calculations from the nutrient status of field as well as higher price of the prescribed fertilizers and amendments.

ConclusionWith the increasing food demand, the concept of sustainable agriculture through judicious management of natural resources (soil being a major one) should be endorsed. Since its inception, the scheme benefitted millions of farmers with the estimation of nutrient status, assessment of crop production potentiality and prescribing precise corrective measures. Building a well-connected lab-to-land knowledge network with greater coverage, robust analysis and report handover, efficient monitoring, and follow up can enhance the utility, applicability, and acceptance of SHC towards the farmers which can accomplish the ultimate objective of food self-sufficiency in light of humongous population growth.

ReferenceSoil Health Card by Department of Agriculture,

Cooperation & Farmers Welfare, Ministry of Agriculture & Farmers Welfare Government of India (2020). Consolidated progress report [data file]. Retrieved from https://soilhealth.dac.gov.in/Home/ProgressReport.



20160

35. Precision Nutrient ManagementDURGESH KUMAR MAURYA1*, GAURAV SHUKLA2

1* M.Sc. & 2 Ph.D. Scholar, Department of Agronomy, Sardar Vallaibh Bhai Patel University of Agriculture and Technology, Meerut 250110

AbstractPrecision nutrient management is one of the main important components of precision agriculture and it manages all the major concerns of improving productivity, profitability, sustainability, and climate change-related instabilities. Soil test-based nutrient managing recommendations have assisted the purpose of improving food grain production but it has not enhanced the nutrient use efficiency elsewhere a certain limit. The modern research is concerned with more towards synchronizing nutrient supply with plant needs.

IntroductionThe ‘Green Revolution’ of the 1960s and 1970s made India self-sufficient in food production. In the past five decades the production of food grains has increased more than three-fold. All this developed because of the availability of high yielding varieties, irrigation, fertilizers, pesticides, and mechanization. According to FAO (2008) report entitled ‘State of Food and Agriculture’ (FAOSTAT, 2009) India has 142 million hectares (Mha) of arable land and has tremendous growth potential for grain production. To connect this potential, we need to accomplish the ‘Evergreen Revolution’. The evergreen revolution means harvesting maximum yields from the available water, land, and other resources, without affecting any ecological or social harm. Precision agricultural technologies and techniques can go a long way in attaining this projected goal. Conserving natural resources and avoiding any ecological or social problems. Precision agriculture has now achieved exceptional growth in the developed countries. Developing countries in Asia have been fairly slow in developing, understanding and adopting precision agricultural practices. The precision agriculture is defined as the science of relating ‘right-input’ at ‘right time’ in ‘right-amount’ at ‘right place’ and in ‘right-manner’ for successful production.

Precision Nutrient Management Tools and Technique

Chlorophyll Meter Leaf Colour ChartOmission Plot Technique Optical Sensor

1. Chlorophyll meterChlorophyll meters are consistent substitutes to traditional tissue analysis as plant N nutritional diagnostic tools. Hand-held Minolta SPAD-502 is the most widely used chlorophyll meter. It rapidly provides an evaluation of leaf N status as chlorophyll

content (Fiebo et al., 1998; Boggs et al., 2003) by holding the un-plucked leafy tissue in the meter used two LEDs (light-emitting diodes) infrared (ë = 940 nm) and light-emitting red (ë = 650 nm). The infrared and red radiations are made to permit through the leaf. A percentage of light is absorbed and the rest is transmitted over the leaf, and a silicon photodiode detector alters it into an electrical signal. The quantity of light attainment the detector is inversely proportional to the quantity of chlorophyll in the track of the light. Leaf chlorophyll content is presented in arbitrary units (0–99.9) and the meter readings are unit less which essential to be adjusted with N content of chlorophyll and leaf greenness.

Two approached used to manage fertilizer N using SPAD meter:A. Fixed threshold value approachThe N Fertilizer is useful whenever chlorophyll meter reading is less than the predetermined threshold value. The SPAD values of the index leaf are observed at 7-10 days’ interval starting from 15 days after transplanting till the beginning of flowering. In-season top-dressing of 30 kg N/ha was suggested whenever SPAD value fell below the critical value of 35 for rice cultivar IR72 grownup in the dry season in the Philippines.

B. Sufficiency index value approachIt is defined as the SPAD value of the test plot expressed as a proportion of the SPAD value of an over-fertilized reference plot. The top-dressed N fertilizer at the rate of 30 kg N/ha whenever sufficiency index was less than 90% up to 50% flowering. The experiment was conducted in Rice grain yields attained for different cultivars that were similar to those obtained in the blanket-N application treatment but with 30 kg less N/ha. The criteria of 90% sufficiency index in direct-seeded rice followed by Bijay-Singh et al. (2006). The sufficiency index value approach saved 50 kg N/ha N fertilizer in comparison to a blanket application of 120 kg N/ha with no reduction in the grain yield.

2. Leaf colour chartIt is a high-quality plastic strip with different shades of green colour ranging from light yellowish green (No.1) to dark green (No.6). The First use of LCC technology in Japan by Furuya (1987). IRRI (1996) reported the developed an improved version of six-panel LCC was developed through the association of the International Rice Research Institute (IRRI) with agricultural research systems of several countries in Asia.



Two approach LCC synchronizing fertilizer (N application with plant needs)A. Real- time nitrogen managementThe LCC mark of the first fully exposed leaf is observed at 7-10 days interval starting from 15-20 days after transplanting till the start of flowering and prearranged amount of fertilizer-N is applied whenever the Colour of rice leaves falls below the critical LCC mark. According to (Bijay-Singh et al., 2003) reported the LCC helped to save 19- 39 kg N/ha in Haryana, 27-56 kg N/ha in Punjab, 30 kg N/ha in Uttarakhand, 20 kg N ha-1 in West Bengal and 30-40 kg N ha-1 in Bihar, as compared to fixed-time blanket N recommendation.

B. Fixed time variable rate dose approachAccording to Bijay-Singh et al. (2012) reported in its place of measuring leaf colour intensity at every 7-10 days interval, LCC can be used to adopt variable rate N dose at fixed growth stages in rice. If the green colour intensity of leaves is higher (for example e” LCC 4), apply less N fertilizer. The fixed time variable rate precision N management strategy in wheat in north-western India conducted a series of experiments to develop by (Varinderpal-Singh et al. 2012). It was created that a dose of only 25 kg N/ha is required at planting.

3. Omission plot techniqueIt is estimated that fertilizer requirements for achieving a yield target. In this practice all the major nutrients are applied excepting the nutrient of interest i.e. omitted nutrient. It is providing an estimate of the native nutrient supply of the soil. For example, if all the nutrients excluding for P are applied in the P-omission plot, then the yield will be limited by the indigenous supply of P.

4. Optical sensorThese techniques measure visible and near- infrared (NIR) spectral reactions from plant canopies to identify the N stress (Peñuelas et al., 1994; Ma et al., 1996). Chlorophyll contained in the picket layer of the leaf controls much of the visible light (400-720 nm) reflectance while reflectance of the NIR electromagnetic spectrum (720- 1300 nm) depends upon the structure of the mesophyll tissues. The Spectral vegetation indices such as the normalized-difference vegetation index (NDVI) calculated as NDVI = (FNIR – FRed)/ (FNIR + FRed)

In this formula indicate are described FNIR means fractions of emitted NIR and FRed means red radiation reflected from the sensed area, According to Peñuelas et al., 1994; Thenkabail et al., 2000; Raun et al., 2001; Báez-González et al., 2002; Kaur et al.(2010)

its afford information about photosynthetic efficiency, productivity potential, and potential yield.

ConclusionPrecision nutrient management observes including use of chlorophyll meter, leaf Colour chart, optical sensors, omission plot technique can help monitor in determining need-based nutrient applications and thus successful nutrient use efficiencies while attaining high yield levels in different crops.

ReferenceBijay-Singh and Yadvinder-Singh 2003. Efficient

nitrogen management in rice-wheat system in the Indo-Gangetic plains. In Nutrient Management for Sustainable Rice-Wheat Cropping System (YadvinderSingh, Bijay-Singh, V.K. Nayyar and Jagmohan Singh, Eds.), pp. 99–114. National Agricultural Technology Project, Indian Council of Agricultural Research, New Delhi and Punjab Agricultural University, Ludhiana.

Bijay-Singh, Gupta, R.K., Yadvinder-Singh, Gupta, S.K., Jagmohan-Singh, Bains, J.S. and Vashishta, M. 2006. Need-based nitrogen management using leaf color chart in wet direct-seeded rice in northwestern India. Journal of New Seeds 8, 35-47.

Bijay-Singh, Varinderpal-Singh, Yadvinder-Singh, Thind, H.S., Ajay-Kumar, Gupta, R.K. and Vashistha, M. 2012. Fixed-time adjustable dose site-specific fertiliser nitrogen management in transplanted irrigated rice (Oryza sativa L.) in South Asia. Field Crops Research 126, 63–69. Varinderpal-Singh, Bijay-Singh, Yadvinder-Singh, Thind, H.S., Gobinder-Singh, SatwinderjitKaur, Ajay-Kumar and Vashistha, M. 2012. Establishment of threshold leaf colour greenness for need-based fertiliser nitrogen management in irrigated wheat (Triticum aestivum L.) using leaf colour chart. Field Crops Research 130, 109-119.

FAOSTAT. 2009. FAO statistic division http://faostat.fao.org.site/575default.aspx#ancor.

Feibo, W., Lianghuan, W. and Fuhua, X. 1998. Chlorophyll meter to predict nitrogen sidedress requirements for short-season cotton (Gossypium hirsutum L.). Field Crops Research 56, 309-314.

IRRI. 1996. Use of leaf color chart (LCC) for N management in rice. Crop and Resource Management Network Technology Brief No. 1. International Rice Research Institute, Los Baños, Philippines.

Peñuelas, J., Gamon, J., Freeden, A., Merino, J. and Field, C. 1994. Reflectance indices associated with physiological changes in nitrogen and water limited sunflower leaves. Remote Sensing of Environment 48, 135-146.



20163

36. Hyperaccumulators in Phytoremediation of Heavy MetalsPALLAVI. T.1* AND SHWETHAKUMARI. U.2

1Ph.D. Scholar; Department of Soil Science and Agricultural Chemistry, UAS, Bengaluru, Karnataka 2Ph.D. Scholar; Department of Soil Science and Agricultural Chemistry, UAS, Raichur, Karnataka *Corresponding Author Email: [email protected]

IntroductionHyperaccumulators are conventionally defined as the plant species that are able to accumulate metals at very high levels (100 times that of non-accumulator species). They have tolerance to exceptionally high amounts of heavy metals and show no significant signs of toxicity. In general, a hyperaccumulator can able to concentrate more than 10 ppm of Hg, 100ppm of Cd, 1000 ppm of Co, Cr, Cu and Pb, 10000 ppm of Ni & Zn. So far, about 400 plants that hyperaccumulate metals are reported from 45 different families. The families dominating these members are Brassicaceae, Asteraceae, Caryophyllaceae, Cyperaceae, Cunoniaceae, Fabaceae, Flacourtiaceae, Lamiaceae, Poaceae, Violaceae, and Euphorbiaceae. With the highest occurrence, the brassicaceae had the largest number of taxa viz. 11 genera and 87 species. These plants are quite varied, from perennial shrubs and trees to small annual herbs. With these significant characteristics, hyperaccumulators are considered as the ideal candidates for phytoremediation of heavy metals. Chaney (1983) was the first to suggest the use of hyperaccumulators for the phytoremediation of metal-polluted sites.

PhytoremediationThe term phytoremediation (phyto meaning plant and remedium meaning to clean or restore) refers to a diverse collection of plant-based technologies that use either naturally occurring, or genetically engineered, plants to clean contaminated environments. Phytoremediation is a term applied to a group of technologies that use plants to reduce, remove, degrade, or immobilize environmental toxins, primarily those of anthropogenic origin, with the aim of restoring area sites to a condition useable for private or public applications. Phytoremediation is clean, simple, cost effective, non-disruptive green technology and most importantly, its by-products can find a range of other uses.

Characteristics of an Ideal Hyperaccumulator for Phytoremediation

� Tolerant to high levels of metal � Profuse root system � Rapid growth rate � Potential to produce a high biomass � Accumulate high levels of the metal in harvestable

parts � Relatively long life

Process of the accumulation of heavy metals in hyperaccumulators

Advantages � In situ remediation � Cost effective � Remove pollutants from soil and reduce their

movement towards groundwater � Sustains the soil properties, enhance soil quality

and productivity � End product can be used as bio-ore of heavy metal

Limitations � Low remediation rate � Time taking (usually oner 3-5 years) � The remediation rate is influenced by the climate,

soil conditions and management practices � Being hazardous, disposal of harvested biomass is

of greater challenge � Hyperaccumulators that are edible are of much

concern as they pave the way for the entry of heavy metals into the food chain.

Effective DisposalThe disposal and management of huge biomass generated by the use of hyperaccumulators for phytoremediation of heavy metal contaminated sites



is of utmost significance and a greater challenge as it creates further pollution problems and environment aesthetic issues. To decrease handling, processing, and potential landfilling costs, waste volume can be reduced by thermal, microbial, physical or chemical means. Also, post-harvest management of generated biomass through advanced techniques like composting

and compaction, combustion and gasification, phytomining and pyrolysis is essential. The biomass generated can also be subjected for extraction biofuel and incineration to further concentrate the bio-ore.

Hyperaccumulators with their Metal Accumulation Capacity

Species Family Metal Accumulation capacity(mg kg-1 DW) Reference

Noccaea caerulescens Brassicaceae Pb 1,700–2,300 Dinh et al. (2018)Alyssum markgrafii Brassicaceae Ni 4,038 Salihaj et al. (2016)Thlaspi caerulescens Brassicaceae Ni 16,200 Koptsik (2014)Brassica nigra Brassicaceae Pb 9,400 Koptsik (2014)Brassica juncea Brassicaceae Se 900 Reevs and Baker (2000)Helianthus annuus Asteraceae Pb 5,600 Koptsik (2014)Tagetes minuta Asteraceae As 380.5 Salazar and Pignata (2014)

Medicago sativa Fabaceae Pb 43,300 Koptsik (2014)Cannabis sativa Cannabaceae Cu 1,530 Ahmad et al. (2016)Cannabis sativa Cannabaceae Cd 151 Ahmad et al. (2016)Pteris vittata Pteridaceae Hg 91.975 Jianxu et al. (2012)Pteris vittata Pteridaceae Cr 35,303 Kalve et al. (2011)Eleocharis acicularis Cyperaceae Zn 11,200 Sakakibara et al. (2011)Thlaspi caerulescens Brassicaceae Cd 3,000 Sheoran et al. (2009)

Haumaniastrum katangense Lamiaceae Cu 8,356 Sheoran et al. (2009)Ipomoea alpina Convolvulaceae Cu 12,300 Reevs and Baker (2000)Haumaniuastrum robertii Lamiaceae Co 10,200 Reevs and Baker (2000)

ConclusionHyperaccumulators have tremendous potential for application in remediation of heavy metal and contaminated sites in the environment. Phytoremediation using hyperaccumulators being an environmentally friendly, low-cost technology for decreasing the heavy metal content of contaminated soils, emerged as a potential alternative to the engineering-based remediation methods. Thus, hyperaccumulators are a vital tool in sustainable management of heavy metal contaminated soils.

ReferencesPandey, V. C. and Bajpai, O., 2019, Phytoremediation:

from theory toward practice. Phytomanagement of Polluted Sites. Elsevier, pp.1-49.

Saxena, G., Purchase, D., Mulla, S. I., Saratale, G. D. and Bharagava, R.N., 2020, Phytoremediation of heavy metal-contaminated sites: eco-environmental concerns, field studies, sustainability issues, and future prospects. Reviews of environmental contamination and toxicology, 249:71-131.

20166

37. Bioavailability of NutrientsPOOJITHA K.1*, PRASHANTH D. V.2*, AND HARSHA B. R.3*

2Ph.D. Scholar; Department of Soil Science and Agricultural Chemistry, UAHS, Shivmogga, 3Ph.D. Scholar; Department of Soil Science and Agricultural Chemistry, UAS, Bengaluru, 1Ph.D. Scholar; Department of Agronomy, GKVK, UAS, Bengaluru *Corresponding Author Email: [email protected].

Bioavailability is also referred bio-accessibility of nutrients “Bioavailability is the degree to which nutrients present in the soil may be absorbed or metabolised by human or ecological receptors or are

available for interaction with biological systems” (ISO 2005).



Impact of Soil Properties on Bioavailability � To be bioavailable molecules must cross a

biological membrane. � In effect this means that the molecules have to

interact with the aqueous phase. � Soil properties which control of availability of

nutrients

– pH,– Organic matter content,– Eh,– Cation exchange capacity (CEC)– Clay minerals and– Oxyhydroxides,

Bio Availability of Metals and Metalloids1. Bioavailability tends to decrease from acid to

neutral through alkaline conditions.2. As pH increases from acid to alkali, metal ions

are more likely to displace protons from exchange sites

3. Soil-metal + x H+ soil-H + metalx+ (Lindsay,1979)4. In contrast to cations, one present as anions

such as arsenate and chromate tend to be more significantly sorbed at lower pH, thus becoming more bioavailable (but also more mobile in the soil) towards more neutral pH conditions.

5. pH dependent and independent charges are important for bioavialability

6. pH dependent charges are dominant in 1:1 than 2:1

7. Fe- and Al-oxyhydroxides are important for retaining bioavailable anions in the soil and Mn oxyhydroxides for retaining cations.

Influence of organic matter on bioavialability of metals and metalloids1. Surfaces of organic matter can have a negative

charge through the dissociation of carboxylic acid and phenolic acid groups and thus can provide exchange sites for cationic metals.

2. However, metals and organic matter also interact to form chelate complexes in which the metal is sorbed to the organic matter through more than

one bond so that a ring structure is formed.3. As an illustration of the formation of chelate

complexes, cadmium is complexed with one deprotonated carboxylate group and copper is complexed with one carboxylate and one neighbouring phenolate group.

4. Different metal cations show different tendencies to form complexes with organic matter.

5. Copper is able to form a strong bound.6. Overall, total metal-binding concentrations to

organic matter decrease in the order copper > nickel > lead > cobalt > cadmium > calcium > zinc > manganese > magnesium

Influence of redox conditions bio-availability of metals1. Elements that can exist in more than one

oxidation state the lower oxidation state ions are more soluble.

2. Under more reducing conditions, the concentration in the pore water often increases.

3. If soils are waterlogged and become anaerobic, oxyhydroxides of Fe and Mn become unstable and dissolve.

4. Precipitation of sulphides can reduce metal availability under reduced conditions.Example: N bio-availability depends on

following chemical transformation

a) Mineralization: Aminization, Ammonification, nitrofication

b) Immobilization:c) Volatilizationd) Denitrification



20170

38. Humus and its Relavences on Soil, Plant and EnvironmentPRASHANTH D. V.1*, HARSHA B. R.2 AND POOJITHA K.3

1Ph.D. Scholar; Department of Soil Science and Agricultural Chemistry, UAHS, Shivmogga, 2Ph.D. Scholar; Department of soil science and agricultural chemistry, UAS, Bengaluru, 3Ph.D. Scholar; Department of agronomy, GKVK, UAS, Bengaluru *Corresponding Author Email: [email protected].

HumusThe decomposition of plant and animal remains in soil constitutes a basic biological process in that Carbon is recirculated to the atmosphere as CO

2 and associated

elements (Plant nutrients). It involves various pools of organic matter such as litter, light fraction, microbial biomass, water soluble organics, stabilised om (humus).

Humic Substance“A series of relatively high molecular weight, yellow to black colour substances formed by secondary synthesis reactions” (Stevenson, 1994).

Soil Humus is also known as “Soil skin/Flesh”. On fractionation, Humus yields- HA, FA and Humin

Humification of organic residues incorporating the soil depends upon

1. Chemical composition2. Soil conditions influencing the activity of soil

micro-organisms.

Influence of humus on soil1. Soil physical structure:

a) Improves soil structureb) Avoids destructive effect on structure

impeded by tillage operationc) Adequate humus level in soil eases seedbed

preparation and tillage operation.d) When humus is lost soil tend to become hard,

compact and cloddye) Enhances aeration, WHC and permiabilityf) Better aggregation, percolation and

infiltration2. Soil warming: Dark colour enhances soil

warming3. Resists erosion by forming granular aggregates.4. Enhances buffering capacity and exchange

capacity of soil.5. Adsorption of pesticides and other organic

chemicals.

Influence of humus on plant nutrient availability1. Direct effects: Plant nutrient supply2. Indirect effects:

a) Provides energy for N2-fixing bacteria.

b) Mineralization of humic substances helps in

stable structures.c) Complexation of Ca2+ esp. in calcareous and

Al and Fe soils and thus releases phosphate ion.

d) Alleviation of metal ion toxicities (Al+3).e) Chelation helps in micronutrient availability.Cropping system is to be considered during

Humus evaluation.

Environmental Significance of Humus � Interacts with metal ions, oxides, hydroxides,

mineral and organic compounds, including toxic pollutants, to form water-soluble and water-insoluble complexes.

� Contributes to reduce toxicity of heavy metals and pesticides.

� Their selective binding capabilities are also exploited for the destruction of chemical warfare agents.

� Humus-based filters have been developed for sewage purification.

� Utilized for sorbing gases, e.g. the removal of waste gases from an animal carcass

Theories of Humus Formation1. The lignin-protein theory (Walksmann-Classical

Theory,1932)2. The Polyphenol Theory (Flaig & Kononova

Concepts- 1964)3. Sugar-amine condensation (Maillard Concept-

1911).



Fractionation of Humus

Fractionation scheme of soil organic matter

Synthesis of Humic SubstancesHistorical review of extraction: Archard (1786) extracted dark amorphous precipitate on acidification in peat with ALKALI, later it was named as Humic acid which was acid insoluble but an alkali soluble. De Saussure introduced term “Humus”. Berzelius isolated 2 light yellow coloured HS from mineral waste and slimy mud rich with Iron oxides.

Synthesis � The change of colour from colourless, brown,

yellow, green, dark green and eventually black solution was a first indication of polymerization reaction taking place, i.e. humic and fulvic acids

formation. � Solid formation of high molecular weight fractions

is also a confirmation of polymerization.

The synthetic procedure of humic substances � The Catechol (6.6g) is reacted with acetic acid

(3.6g) in a 1.0L glass beaker. � The pH of the reaction medium (water and

catechol) is adjusted to 5.50 using 1.0M NaOH and/or 1.0M HCl and allowed to stand in the dark at 25°C (±0.1°C), under constant stirring at 3,000 rpm, which provides sufficient homogeneity.The polymerization reaction is carried out for 30

days while pH has to be maintained at 5.50 throughout the synthesis.

� The colour of the medium slowly turns into black as an indicator of the polymerization reaction. At the 31st day, the reaction medium is transferred to dialysis membranes.

� A series of dialysis at 500, 1,000 and 10,000 MWCO, each for 7 days was performed in order to have calibrated fractions.The fractions obtained were then deep frozen,

lyophilized, ground and stored at 4°C.

ConclusionHumus is dark amorphous substance which is resistance for further degradation consist of various form of nutrients which act as a source for the nutrient in soil and increases physical, chemical and biological properties of soil in turn maintain good soil health.

20178

39. Role of Sulphur in Indian SoilsAMBIKA PRASAD MISHRA

Ph.D. Research Scholar, Department of Soil Science & Agricultural Chemistry, College of Agriculture, OUAT, Bhubaneswar – 751003 *Corresponding Author Email: [email protected]

Sulphur (S) balances in agricultural soils have become a significant concern for the agriculturists all over the world because they are mostly negative. Such situation, i.e., declining S levels in soils have been attributed to strict environmental rules on industrial S emissions, use of high analysis S-free fertilizers and high yielding varieties, intensive cropping, and limited or no use of organic manures and S-containing pesticides. Tripathi claimed that at least 57 million ha out of 142 million ha arable land in India is deficient in S. Several researchers have reported S deficiency in various states of India, viz., Uttar Pradesh, Uttarakhand, Odisha, Madhya Pradesh, Maharashtra, Jharkhand, West Bengal, Andhra Pradesh, and Karnataka. Similarly, the deficiency has been observed in many regions of the world due to continuous depletion of native S reserves. Thus, the importance of S was recognized very quickly, and S-containing fertilizer products were introduced

into the markets.Sulphur is the fourth primary plant nutrient

required for the normal growth of plants, and it plays an important role in many plant processes; which indicate that plant metabolism is dependent upon S and its deficiency will cause primary metabolic impairment. Plant S concentrations are found to be lower than nitrogen (N), but quite similar to that of phosphorus (P) Ali et al., (2012). It is essential for the synthesis of amino acids (cysteine and methionine) which are the basic structural units of protein molecules and constituent of several enzymes, chlorophyll, oils, and vitamins. It regulates the activity of nitrate reductase in plants, and also helps in microbial fixation of atmospheric N (Lucheta, 2012). The behavior and reactions of S in the soil are very similar to those of N which are mainly dominated by the organic or microbial fractions of soil. Its deficiency often becomes



a major hindrance for the sustainable growth and productivity of field crops. Successful crop production of not only of oilseeds, pulses, vegetables, and forages but also of many cereals are dependent on S nutrition.

Generally, the use efficiency of S fertilizers is very low (8-10%). Until now traditional S fertilizers, viz., ammonium sulphate, single super phosphate (SSP), gypsum, etc. were common in use, while S fertilizers like elemental S (S°), bentonite S, micronised S, etc. and their advanced formulations are becoming popular nowadays. The inorganic fertilizers containing S as sulphate (SO

4 2-) and S° fall in the category of

conventional and advanced S fertilizers, respectively.Elemental S came into the demand because of high

concentrations of S (70-100%), negligible leaching and run-off losses, continued residual effects on the S nutrition of the subsequent crop, and low transport and application costs (for it is 100% S). Sulphate fertilizers provide S to plants quickly, but they are susceptible to leaching losses.

Plant takes S in SO4

2- form, so the S° fertilizers must be converted into that form through the process of oxidation which is mediated the by S-oxidizing microbes. A similar process is also required for bentonite and micronised S fertilizers for solubilisation of S. These fertilizers have been termed as slow-release fertilizers, and they have the advantage of the long-term supply of S to crops.

The Requirement of Sulphur for Crop NutritionSulphur plays an important role in improvement of yield and quality of crops. It is linked with N metabolism, and its application increases the uptake of N by plants. Besides N, it also enhances the uptake of other beneficial nutrients like phosphorus, potassium, and zinc, and checks the uptake of toxic elements like sodium and chlorine. Plants deficient in S have less resistance to biotic and abiotic stresses. Moreover, the primary and secondary metabolism involving amino acids, carbohydrates, glucosinolates, and biosynthesis of many other secondary compounds are moderated in plants during S stress (Saito,2004). Visual symptoms of S deficiency include chlorosis on younger leaves and reduced plant growth (premature defoliation, thin and woody stem, reduced leaf size, stunted growth, etc.); and its toxicity symptoms include chlorosis, interveinal necrosis, mottling in young leaves, inhibition of apical growth, bluish green appearance of older leaves, busy appearance of lateral branches, and ultimately reduction in growth.

The requirement of S nutrition varies with the type of crops. It is generally in the order of Cruciferae > Leguminosae > Gramineae. Oilseed crops are known

to deplete the S content of soil as their uptake for production of seed is very high. Sulphur required to produce one ton of seed is about 3-4 kg for cereals, 8 kg for pulses, and 12 kg for oilseeds. Walker and Booth, 1992 estimated crop removal of S for oilseed rape is 20-30 kg ha-1, but for cereals, it is 10-15 kg ha-

1. Plant S concentrations vary between 0.1 and 0.5%. The content is generally high during vegetative growth stages compared to maturity. The N: S ratio is also an important factor which influence the S requirement of plants since both N and S are closely linked in synthesis of protein, addition of N must be considered in scheduling S fertilization. Generally, it is established that one part of S is required for every 15 parts of N, and their ratio lies in the narrow range of 15:1. However, fertilizer recommendation also depends on climatic conditions, locations, soil types, and cultivars (Kruse et al., 2007).

The production of oilseed crops and their quality is significantly influenced by supply of proper S nutrition. Quality attributes of oilseeds like oil content, glucosinolate concentrations, protein concentrations, etc. were found to increase with appropriate S application. Similarly, the growth and development of many crop species are affected; and are responsive to S supply. Salvagiotti et al. observed high nitrogen use efficiency in wheat as S addition enhanced the N uptake. Biological N

2 fixation and consequently higher

dry matter accumulation in legumes is influenced by S fertilization because of increased nodulation and better root growth.

ReferencesTripathi N. Role of FCO in promoting quality secondary

and micronutrients. Fertiliser News. 2003; 48: 111-114.

Ali A, Arshadullah M, Hyder SI, Mahmood IA. Effect of different levels of sulfur on the productivity of wheat in a saline sodic soil. Soil and Environment. 2012; 31(1):91-95.

Lucheta AR, Lambais MR. Sulfur in agriculture. Revista Brasileira de Ciência do Solo. 2012; 36:1369-1379.

Saito K. Sulfur assimilatory metabolism. The long and smelling road. Plant Physiology. 2004; 136:2443-2450.

Walker KC, Booth EJ. Sulphur research on oilseed rape in Scotland. Sulphur in Agriculture. 1992; 16:15-19.

Kruse C, Jost R, Lipschis M, Kopp B, Hartmann M, Hell R. Sulfur-enhanced defence: effects of sulfur metabolism, nitrogen supply, and pathogen lifestyle. Plant Biology. 2007; 9:608-619.



20198

40. Soil Sampling and Testing: A Tool for Soil ManagementROOHI1 AND HARDEEP SINGH SHEORAN2

1Regional Research Station, CCS Haryana Agricultural University, Karnal-132001, Haryana 2Deptt. of Soil Science, CCS Haryana Agricultural University, Hisar-125004, Haryana *Corresponding Author Email: [email protected]

Some of the soils were already deficient in nutrient and some were nutrient rich earlier. But with intensive cultivation, soils which were having sufficient level of nutrients gets exhausted and there is a need to sustain nutrient status by adding nutrient back to the soil reserves. The quantity of nutrient removed or added we can’t tell exactly just by seeing the soil. Soil testing is a tool to analyze the soil physical as well as chemical status and could help to analyze the amount of inorganic fertilizer that need to be added for enhancing crop yield. This will not only help to improve yield but can also diagnose the effect of toxic metals on environment. The objective of soil testing is to measure soil nutrient status which further can answer following questions of farmers:1. Does my soil is deficient in nutrient?2. Does my crop need fertilizer?3. How much fertilizer do I need to add?4. What type of fertilizer should I use?

This not only answers farmer’s questions but can also help them to reduce the cost of cultivation and helps to select crop based on the soil type. As most of the farmers are unknowingly adding fertilizer in

excess, it effects the farmer’s economics, soil health and environment. So, soil testing is widely adapted tool. It involves collection of soil sample, preparation, analysis (Chemical and Physical parameters), interpretation of analysis results and fertilizer recommendation based on the soil status and crop requirement.

Time of collection: Usually after the harvest or just before sowing of the crop is the right time of sampling. In the standing crop, if plant show deficiency symptoms then sample should be collected in between the rows. So that proper preventive measure can be taken up by farmers. Sampling is recommended once in three years for all soil. In case of intensive cropping, it’s recommended every year.

Collection of soil sample: First clean the surface and then take the sample using auger or spade. Sampling depth differs based on crop (Table 1) Take samples from several locations in a zig-zag pattern.

� Using auger: Drive auger up to plough layer (0-15cm) and take out the sample

� Using Spade: Make a ‘V’ shape cut up to 15cm and take out the sample from upper thin slice of soil from to bottom at the sampling spot.

FIG 1. Collection and preparation of soil sample (Source: Parewa et al., 2016)



TABLE 1. Depth for sampling for different crops

Crop Depth (cm)Shallow rooted crop (Paddy, millets) 15cmDeep rooted crops (Sugarcane, cotton, vegetable)

30cm

Horticultural crops Three different depths i.e., 30,60, 90 cm from 5 different sites

Soil Sample PreparationMix the collected representative sample thoroughly and remove pebbles, stones, gravels. Reduce the volume of sample to 0.5 kg by quartering process. In quartering process, first sample is divided into 4 equal parts and discard the opposite portion and remaining portion should be mixed together and repeat the same until we are left with 0.5kg of sample. Store the sample in polythene bag and label it with important information (Fig 1).

Precaution during Sampling1. Sampling tools or polythene bag (for storage)

should be clean properly.2. Avoid mixing of sampling of field having different

soil color, slope, drainage or different management was followed.

3. For acidic and saline affected soil, take samples which are more prone to symptoms. Avoid sampling after irrigation, fertigation or manuring practice was done.

4. Avoid sampling near to roads, bunds, wet spot, trees or dead furrows.

5. For agricultural crops, Sampling depth taken should be 15cm. For deep rooted crop, it is 30cm and for horticultural crop, it can go up to 90-120cm.

6. Store the sample away from fertilizers, herbicide, pesticide etc.

ReferenceParewa, H.P., Jain, L.K., Mahajan, G.R. and Bhimawat,

B.S. (2016). Soil Health Card: A Boon for the Indian Farmers. Indian Journal of Plant and Soil, 3(2): 77-81.

HORTICULTURE

20123

41. Salicylic Acid: A Potential Phytochemical for Sustainable Fruit ProductionFARHANA KHATOON, DR HIDAYATULLAH MIR AND KHUSHBOO AZAM

Department of Horticulture (Fruit and Fruit Technology), Bihar Agricultural University, Sabour, Bhagalpur-813210, Bihar

Acetyl salicylic acid (ASA) is a derivative of salicylic acid (SA) which when applied exogenously, undergoes spontaneous hydrolysis and is converted to SA (Popova et al.,1997). SA is a natural phenolic compound involved in regulation of many processes in plant growth and development and also known for its induction of plant defense against biotic and abiotic stress. Exogenous application of SA may also induce the expression of pathogenesis-related protein and establish systemic acquired resistance. White willow is a natural source of salicylic acid and SA can be extracted from bark of the tree. SA is a potential growth regulator for improving crop yield under deficit soil water condition.

Role of Salicylic Acid on Fruit Crop1. Flowering behavior: salicylic acid acts as

chelating agent, induces flowering and also effects the florigenic activity by functioning as endogenous plant growth regulator. It has been reported to influence flowering in olive (El-Razek

et al., 2013). Salicylic acid promotes flowering and has been found to increase the number of male and hermaphrodite flower per panicle by increasing the flowering stimulus in mango (cv. kesar) (Ngullie et al., 2014).

2. Post-harvest storage life: Post-harvest application of methyle salicylic acid decrease the ACC oxidase and ACC synthase activity and helps in retaining the firmness of kiwifruit during storage and prevent it from softening (Fattahi et al.,2010). SA acts as anti- senescence and its exogenous application in ponkan mandarin reduce the post-harvest fruit decay. Its treatment also reduces the susceptibility to chilling injury in pomegranate and avocado.

3. Fruit ripening: SA delays fruit ripening process in banana by inhibiting the ethylene biosynthesis pathway and it reduces the activity of major cell wall degrading enzymes like cellulase, polygalacturonase and xylanase (Srivastava et al.,2000) Some findings reveals that exogenous



pre and post-harvest application of salicylic acid on sweet cherry decreases the endogenous ethylene production and its treatment also delay the ripening process in guava.

4. Plant growth and development: SA influences the biochemical and physiological processes of plant by enhancing ion uptake, membrane permeability, heat production and growth and development of plant. Strawberry treated with proper dose of SA showed increased leaf area, fresh root and shoot weight and leaf nitrogen concentration (Jamali et al 2011).

Whiite Willow (Salix slba) Function of Salicylic Acid in Plants

5. Photosynthetic activity: Salicylic acid have an important role in regulation of various metabolic processes, water status in plants, activity of enzymes such as carbonic anhydrase and Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase), chlorophyll and carotenoid contents, also affect leaf and chloroplast structure and stomatal closure. Spray of SA increase the chlorophyll content in strawberry which enhance the photosynthetic rate in plant.

6. Fruit yield: Salicylic acid enhances yield of the crop by increasing photosynthetic activity in leaves and translocation of more photoassimilates to fruits. Mango (cv. Kesar) trees sprayed with 2000 ppm salicylic acid recorded higher number of fruits per tree) (Ngullie et al., 2014). Foliar application of SA on strawberry cultivars increased early, late, and total fresh weight yield per unit area. SA spray also improves the yield in

olive tree.7. Fruit quality: SA decreases polyphenol oxidase

activity and improve anthocyanin pigments of the fruit pericarp. Application of SA was found effective to overcome pericarp browning in litchi (Kumar et al.,2013). Salicylic acid treatment in mango cv. Kesar resulted in significantly highest total soluble solids (ºBrix), reducing sugar (%) and minimum titrable acidity (%).

8. Biotic stress tolerance: SA is an important mediator of the plant defense response to pathogens. Activities of defense enzymes of POD (peroxidase), PAL (phenylalanine ammonia-lyase), chitinase and b-1,3-glucanase in the young fruit were notably enhanced by SA sprays on the trees. Disease incidence and development in the pear fruit inoculated with P. expansum were significantly reduced by SA treatment during fruit development stage which provide protection against post-harvest diseases.

9. Abiotic stress tolerance: exogenous treatment of SA on plant provide resistance against drought and salt stress. SA improves leaf water status under water deficit condition by closing stomata. Plant under abiotic stress generate reactive oxygen species (ROS) which cause damage to cell components. Exogenous SA activates antioxidant defense system to scavenge ROS and help to tolerate stress. In strawberry SA spray increases the activities of antioxidant enzymes (POD, catalase (CAT) and ascorbate peroxidase (APX) and improved plant drought stress tolerance (Ghaderi et al.,2015) SA spray provide osmotic adjustment to the plant by accumulating different osmolytes such as sugars, sugar alcohol and proline and provide the ability of plant to survive under high salt condition. SA increases fresh shoot weight, dry shoot weight, fresh root weight, dry root weight, and chlorophyll content in strawberry plant under salinity stress (Joseph et al., 2010).

ConclusionSA is very much effective to improve vegetative growth and biological yield of the plant. SA play an important role in pigment biosynthesis and enhances photosynthetic rate, SA is having florigenic activity, inhibits ethylene biosynthesis and increases storage life, act as an antioxidant and reduces browning in fruits, SA influences various enzymatic activities and improve quality attributes of different fruit crops with increased stress resistance. New molecules like SA are ecofriendly approach to increase the treatment of crop production, so promotion of these molecule is highly required for achieving the dream of sustainable fruit production.

ReferencePopova, L., Pancheva, T., & Uzunova, A. (1997). Salicylic

acid: properties, biosynthesis and physiological role. Bulg. J. Plant Physiol, 23(1-2), 85-93.

Ngullie, C.R., Tank, R.V. and Bhanderi, D.R. (2014). Effect of salicylic acid and humic acid on



flowering, fruiting, yield and quality of mango (Mangifera indica L.) cv. KESAR. Adv. Res. J. Crop Improvement. 5 (2): 136-139.

Fattahi, J., Fifall, R., & Babri, M. (2010). Postharvest quality of kiwifruit (Actinidia deliciosa cv. Hayward) affected by pre-storage application of salicylic acid. South Western J. of Hortic. Biol. and Environ, 1(2), 175-186.

Srivastava, M. K., & Dwivedi, U. N. (2000). Delayed ripening of banana fruit by salicylic acid. Plant science, 158(1-2), 87-96.

Jamali, B., Eshghi, S., & Tafazoli, E. (2011). Vegetative and reproductive growth of strawberry plants cv. ‘Pajaro’affected by salicylic acid and nickel.

Joseph, B., Jini, D., & Sujatha, S. (2010). Insight into the role of exogenous salicylic acid on plants grown under salt environment. Asian Journal of Crop Science, 2(4), 226-235.

Ghaderi, N., Normohammadi, S., & Javadi, T. (2015). Morpho-physiological responses of strawberry (Fragaria× ananassa) to exogenous salicylic acid application under drought stress.

Kumar, D., Mishra, D. S., Chakraborty, B., & Kumar, P. (2013). Pericarp browning and quality management of litchi fruit by antioxidants and salicylic acid during ambient storage. Journal of food science and technology, 50(4), 797-802.

20146

42. Seed Production Technical Process in Special Reference CucurbitsDR. RAKESH KUMAR MEENA AND TARUN NAGAR

Assistant Professor, Career Point University, Kota

IntroductionSeed quality has a special importance in agriculture because despite the climatic conditions required by our crops, the average production of almost all crops is very low. The main reason for this is the frequent use of low-quality seeds by the farmers of the state. The seed is considered to be of excellent quality, having 100% genetic purity, free from seeds of other crops and weeds, free from the effects of diseases and pests, which is full of vigor and strength and has high germination capacity. In which the field is good and the yield is good. Livelihood of about 70 percent of the population of the country and state is based on agriculture. Whose desired improvements in economic and social status are possible only through strengthening of agriculture. The high-quality seeds of these growing species have a high position in sustainable agricultural production. Temperature exceeds more than 300C stability of gynoecious sex expression is affected. Homozygous gynoecious lines are maintained by using GA3 (1500 ppm) or at silver nitrate (200-300 ppm) or silver thiosulphate (400 ppm) to induce staminate flowers when sprayed at two to four true leaf stage. Maintenance of the gynoecious lines has been possible through exogenously applied silver nitrate (Kalloo and Franken, 1978). By providing only certified seeds of the latest species to farmers, production can increase by 15 to 20 percent. Munger (1979) reported that the gynoecious lines Gy 14, SR551F, Gy 3, Gy 57 and Table green 68 are the most suitable to produce F1 hybrids in slicing and pickling cucumbers in temperate regions.

The leaves have blood sugar improvement properties (Rai et al. 2008). Achieving homozygosity in male and female parents is difficult. Select male parents with potential vigour, more number of male

flowers, internodes, and disease resistance attributes to be used as testers. Clones (female parent) × Testers (male parent) F1 Selection for superior seedlings and subsequent clonal generations. Pointed Gourd Hybrid (CHES-38 × CH-2) developed at CHES, Ranchi. (V.S.R. Krishna Prasad et al. 2006)

TABLE: Foliar Spray of PGRs to Induce Increased Proportion of Pistillate Flowers

PGR Conc (mg/l) CucurbitsCycocel (CCC) 250-500 effective in cucumberEthephon (CEPA) 150-200 Most cucurbitsGibberellic Acid (GA) 150-200 WatermelonIndole acetic acid (IAA)

10 Snake gourd & bitter gourd

NAA 20-200 Cucumber, melons & gourds

Maleic hydrazide (MH)

25-10050-150

Cucumber, muskmelon, Bottle gourd, ridge gourd

Seed Production Technical ProcessIn this method, a plan is produced in accordance with the seed standards in scientific ways so that the work of production, processing, storage and distribution is executed effectively and the quality of the seed remains till the seed is sown. This process has the following characteristics.1. Genetically and physically pure base seeds are

used.2. Advanced agronomic methods and crop protection

are adopted.3. Specific separation distances from sources of

genetic or physical contamination are taken care.



4. Seeds of unsuitable plants are extracted from the crop in time.

5. Weeds and plants of other crops are also expelled from time to time so that these seeds do not mix in the crop seeds.

6. The diseased plants are also removed before spreading the disease.

7. Special care is taken in seed crop harvesting, mowing, mowing, cleaning etc. so that there is no mechanical damage and mixing.

8. Special attention is paid to prevent pest, disease infection, etc. during storage.

9. To check genetic and physical purity. In addition to tests are done, germination test, humidity test etc. are also done.

10. Seeds are processed with special vigilance so that the quality of the seeds remain consistent with the standards.

11. The processed seeds are filled in appropriate bags and sealed by attaching the certificates.

12. Seeds are stored at a minimum temperature and humidity to prevent disease and Keep seeds safe from pests and germination capacity is not affected.

QualityThere is a provision of seed certification to ensure quality of seeds at desired level. Authentication of parent seeds is done by a committee constituted, while the responsibility for certification of basic and certified seeds is with the seed certification agency of the state. The authentication process is completed in the following step.

Seed verificationFor the production of basic and certified seeds, it is necessary to use breeder and base seeds respectively. The production of seeds of the same category is allowed under special circumstances. At the time of inspection, the seed certification agency verifies the seed source by bill, store receipt and tag.

Crop inspectionTwo observations are necessary at the time of flowering

and harvesting. At the time of inspection, the seed crop should not contain undesirable plants. The crop should also be weed free. Counts are taken from place to place in the field at the time of inspection. The number of counts depends on the area of the field and the number of plants a count. If the number of plants planted in the count exceeds the prescribed standard, the crop is canceled.

Laboratory testAfter legislation, judgments from each lot are taken and sent to the laboratory for testing. Parent seeds are tested in the university and the basic and certified seeds are tested in the seed certification agency’s laboratory. If no judicial seed is found to conform to the standard, then it is canceled. The basic and certified seeds are tested in the laboratory of the seed certified institution. If no judicial seed is found to conform to the standard then it is canceled.

TaggingAfter legalization, the seeds are filled in bags of such size that one acre of seed should be brought in it. The golden yellow tag on the parent seed is provided by the seed certification agency, the white and blue tags on the respective breeder and the base and certified seeds respectively.

References1. Krishana Prasad VSR, KE Lawande and V Mahajan.

(2006). Performance and diversity pattern in the land races of Allium cepa L. Indian J. Plant Genet. Res.

2. Rai PK, Jaiswal D, Singh R, Watal G (2008). Glycemic property of Trichosanthes dioica leaves. Pharm Biol 46:894–899.

3. Kalloo and S. Franken. (1978). Chemical induction of staminate flowers in four determinate gynoecious lines of pickling cucumber. Gartenbauwissenschaft 43(6) 280-282.

4. Munger, H.M. (1979). A summary of cucumber released from Cornell breeding program. Veg. Improv. Newsl. 21: 3-4.

20151

43. Grafting in Solanaceous and Cucurbitaceous Vegetable CropsUMESH, B. C. AND JEEVITHA, D.

Ph.D. Scholars Dept. of Horticulture UAS, Dharwad-580005 *Corresponding Author Email: [email protected]

Soil borne pathogens can be effectively controlled by using an alternative approach called grafting which is a novel technology used in many vegetables now a days. Grafting as a technology for the commercial production

was later on adopted by many countries in Europe, Middle East, Northern Africa, Central America and other parts of Asia. For the production of many fruit-bearing vegetables these seedlings besides providing



resistance against biotic and abiotic stresses, increase the yield of the cultivars. This technique is considered eco-friendly for sustainable vegetable production because the resistant rootstock reduces dependence on agrochemicals. Grafting improves quality of the plant and is used to induce resistance against low and high temperatures. Scion quality is directly contribute to growth and yield parameters. Due to high post graft mortality of seedlings, this technology is still in infancy in India. For its commercial application in India, sharpening of grafting skills need to be standardized.

HistoryJapan and Korea are the first countries to start the grafting programme as early as 1920s with watermelon (Citrullus lanatus) grafted onto pumpkin (Cucurbita moschata) rootstock. Soon after, watermelons (Citrullus lanatus) were grafted onto bottle gourd (Lagenaria siceraria) rootstocks. Brinjal (Solanum melongena) was grafted onto scarlet eggplant (Solanum integrifolium Poir.) in the 1950s. Later, grafting was introduced to North America from Europe in the late 20th century and it is now attracting growing interest, both from greenhouse growers and organic producers. Grafting of vegetables was originated as a random experiment in Korea and Japan. In temperate regions of the world summer season vegetables are grown under green houses. With the ban of fumigant fungicides and nematicides since 1970s in European countries, grafting of susceptible scions on resistant rootstocks is become an effective alternative against soil borne pathogens. Grafting helps in production of healthy vegetables because here the usage of chemicals was less and also grafted plants less prone to disease and pests. Nearly45 million grafted tomato plants are produced in Spain and over 20 million ton in France and Italy. Even though grafting originated in East Asia, it didn’t get attention among the growers of South Asia. Grafting was popularized among cold countries where the year round production is possible under green house.

Key for Success � Selecting suitable scion and rootstock is important

in this grafting procedure � Grafting clips should also be selected according

to the size. Too large clips cannot hold the grafted union together or too small clips gives too much pressure and may deform the union

� Make sure that prepared rootstocks do not have auxiliary bud at the base of remaining

cotyledonary leaf. Rootstock grows -out in the field is a must avoid situation

Methods of Vegetable Grafting � Tongue Approach or Approach grafting � Hole Insertion or Top Insertion grafting � One Cotyledon or Slant or Splice grafting � Tube grafting � Pin grafting � Automated grafting � Robot grafting

Before Grafting � Expose the scions and rootstocks to sunlight for 2

to 3 days before grafting. � Drying of the potted soil where the scion and

rootstock grow by controlled watering to avoid spindly growth.

� Scion and rootstock with similar diameters are important to increase the survival rate

After Grafting � Keep 100% RH for 3 days and then gradually

reduce the humidity. � Keep the light intensity at 3-5 k lux

Disadvantages of Grafting � More labour is required � High cost of grafted seedlings � Fruit quality could be down not all rootstocks are

good � Special care is required � Additional charge of transplanting in case of

cucurbits � It can be overcome by the heavy yielders

Conclusion � Grafting provides a site-specific management tool

for soil borne diseases. � It reduces the need for soil disinfectants and

thereby environmental pollution. � Grafting technology has a potential in promotion

of cultivation in non-traditional and fragile agro-eco system

� Grafting is a rapid alternative tool to the relatively slow breeding methodology aimed at increasing biotic and abiotic stress tolerance of fruit vegetables.

� Since grafting gives increased disease tolerance and vigor to crops, it will be useful in the low-input sustainable horticulture of the future.



20152

44. Hydroponics: The Future of Fruit FarmingHIDAYATULLAH MIR1 AND PREETI SINGH2

1Department of Horticulture (Fruit and Fruit Technology), Bihar Agricultural University, Sabour, Bhagalpur-813210 2Division of Fruits and Horticultural Technology, ICAR-IARI, New Delhi-110012

The ever-growing population, urbanization and decreasing cultivable land has posed a challenge in front of farming community to provide quality nutritive food to the people. These challenges have led to the development of many technologies by the researchers, and hydroponics is one among them, which aims at catering the growing demand for quality horticultural produce, mainly vegetables and fruits. Hydroponics is basically a technology of growing plants in soilless medium containing mineral nutrients in water. Hydroponics cultivation of horticultural crops is gaining popularity among the farmers and entrepreneurs mainly in urban areas as it requires soil-less media, limited space and provides production throughout the year. It has become one of the most popular agricultural technology which aims to overcome global food, land and water shortages and produce quality and nutritious food with limited resources. This technology is becoming popular not only among the progressive farmers but also among urban and peri-urban households as it provides round the year nutritive and safe horticultural produce and also utilizes kitchen garden, rooftop and balcony in urban areas.

Hydroponics technology is quite popular in cultivating high value vegetable and flower crops, but growing of fruit crops possess several difficulties owing to its large size and perennial nature. Fruit trees require large land area for cultivation and soil for support and growth. But there are fruit crops like strawberry, blueberry, which are similar to vegetable and flower crops in growth habits, are smaller in size and have a short life span which makes them suitable for hydroponics cultivation. Other than cultivating these crops in hydroponics facility one can also think about raising fruit seedlings of the woody perennial fruit trees as they will require less area and less time. Some of the advantages of hydroponics technology are- optimum control of nutrition in the media along with the other environmental parameters, helps in avoiding soil-borne pathogens and diseases, leads to enhanced water and nutrient use efficiency and ultimately results in high yield and quality products with year-round production. Large scale adoption of this technology is limited due to factors like high installation and operational cost, intensive and skilled labour.

Techniques of Hydroponics: There are basically three techniques of hydroponics systems-

1. Nutrient Film Technique (NFT): In this technique the nutrient solution is recirculated through the

roots of plants placed in channels made up of wood, tubes, plastic or concrete and the nutrient solution is either pumped through it or passed under gravitation reaching the root system effectively. This technique is effective only for the plants with large root system.

2. Deep Technique Film (DFT): In this technique plants are grown with their roots submerged in floating nutrient solution on a flat table. This method is effective for growing plants with short root system.

3. With Substrate: In this technique, various materials of mineral and organic origin such as coco peat, pine bark, coconut fibre, vermiculite, perlite, rock wool, sand, etc. are used in structures such as tubes and pots to support plant growth and development. These inert material media has high water holding capacity and porosity. In this system, nutrient solution passes through the media and is subsequently drained and collected in collection tank which is re-circulated again and again. This system is popular for strawberry cultivation around the globe.Hydroponics in Fruit Production:

Herbaceous, small structured and annual fruit crops like strawberry, blackberry, blueberry, and raspberry are suitable for production under the hydroponics system. Strawberries and blueberries cultivation is considered best under hydroponics system because of their requirement of acidic growth medium which can be easily controlled and maintained under hydroponics. In temperate and semi-arid regions, strawberry is largely cultivated under hydroponic conditions in greenhouses, because of controlled environmental conditions and lower water requirement. Hydroponics and greenhouse technology together results in higher yield, forcing possibilities, and better pests control thus, reduces the usage of water and chemicals. Among all the fruit crops, it is strawberry that is widely cultivated under hydroponics condition and has resulted in quantitative and qualitative increase in yield than soil cultivation.

Strawberry is the most popular fruit grown under hydroponics system and it allows commercial production of fruit round the year. For its commercial cultivation under hydroponics, NFT is the most desired technique with free draining media like perlite and vermiculite, as strawberry is more susceptible to crown rot and fruit rot caused due to inadequate drainage and waterlogging. For off-season production, day neutral



cultivars should be planted in free draining, clean and sterilized media. The developing fruitlets should be isolated from the damp media using a white plastic mulch as it may result in fruit rot (Morgan, 2000).

Blueberry cultivation and its expansion is limited by the propagation difficulties. Hydroponic system tends to overcome this limitation and allows better plant growth with enhanced yield.

Banana is another fruit plant which can be grown successfully in hydroponics. Being herbaceous and monocarpic it becomes more suitable for a hydroponics system. Dwarf Cavendish banana was grown in high light intensity with high potassium supplement (Morgan, 2000).

Hydroponics in Production of Fruit Seedlings: The initial development phase of seedlings directly affects the orchard productivity, so opting for better seedling production method can result in better productivity. Seedlings of woody plants such as grape, citrus, peach, guava and pear have been successfully produced using hydroponics. Hydroponics could be a useful technology to raise fruit crops and seedlings but further research is required to develop a better

and efficient system along with focus on optimization of nutrient solution and substrate used for raising the seedlings.

Conclusion: Hydroponics to a greater extent has the potential to overcome the limitations of soil-based culture and climate change. For hydroponics technology to be successful in growing fruit crops and seedlings, development of low-cost techniques and structures is of utmost importance as it will involve lower installation and operational cost along with easy maintenance and less labour. In addition to this optimization of perfect system for specific crop is also important. As technologies like hydroponics have the potential for doubling farmers income so to promote the hydroponics cultivation of horticultural crops several state governments and agencies like National Horticulture Board (NHB) are providing subsidies to the farmers for the capital cost involved.

ReferenceMorgan, L. 2000. Grow your own hydroponic

strawberries. The best of The Growing Edge. New Moon Publishing, Inc. Oregon. USA, pp. 99-102.

20208

45. Hydroponics: Advanced Technique of Vegetable Crop ProductionKHYATI SINGH1* AND MUKESH KUMAR MEHLA2

1Department of Vegetable Science, CCS Haryana Agricultural University, Hisar, Haryana-125004 2Department of Soil Water and Engineering, Maharana Pratap University of Agriculture and Technology, Udaipur, Rajasthan-313001 *Corresponding Author Email: [email protected]

IntroductionHydroponics is an advanced technique that has recently reached to the stage of commercial application in India. In this technique, plants are grown in nutrient solutions where inert medium like peat moss, vermiculite, gravel, saw dust, coir dust, rockwool, etc. may or may not be used to give mechanical support to the plants. The term hydroponics is taken from Greek words, hydro (=water) and ponos (=labour) and that means water work (Sharma et al. 2018). When plants are grown in fields, they uptake nutrients from soil, while in hydroponics growing, nutrient solution must be added in water. In simple words, hydroponics means cultivation of plants in the absence of soil, using water containing fertilizers as nutrient source as growing medium. Water is as good as soil for providing nutrients to plants.

Why we need Hydroponics?There are many reasons which contribute to increasing emphasis on hydroponics in India such as the quality of soil has been compromised due the modern agricultural practices over the years. The cultivable lands are

declining with the great rise in industrialization and urbanization. The rising temperature, hot winds, droughts, and unpredictable weather conditions have negative effect on the plant growth and yield. The polluted water sources and poor water management are another reason for adopting soil less cultivation instead of soil cultivation. There are many high value vegetable crops are successful under hydroponics system like tomato, lettuce, cucumber, watermelon, spring onion, pepper, spinach, coriander, potato, etc. Salad crops like basil, swiss chard and microgreens which require low fertiliser inputs can also be grown (Bulgari et al. 2017).

Basic requirements for HydroponicsThe growing popularity hydroponics techniques is due to its proficient management of crop production and resources. For the good production of vegetables year after year, one must be skilled with the fundamentals of hydroponics like crop, nutrient medium and water. It requires attending the plants regularly; hence the availability of labour is must. It should be established at suitable location and the source and availability of



clean water should be ensured before adopting the system. As concentration of ions in water change with time, temperature, pH, electrical conductivity, and dissolved oxygen should be the measured adequately for proper management of nutrients in water. The nutrient balance can be improved by periodic analysis of nutrient ratios and adjustment of nutrient solutions (Son et al. 2016).

Hydroponic Structures and their OperationHydroponic system is based on the principle of recycling and reuse of nutrient solution and supporting media. The different types of structures are explained as below:

Wick systemIt is the simplest structure which does not require any pumps, electricity, and aerators (Shrestha and Dunn, 2016). It is a passive system as it does not require moving parts. Plants are cultivated in grow tray filled with absorbent medium such as coir, perlite and vermiculite and a nylon wick from plant roots is passed into nutrient solution reservoir. Plants uptake water and nutrient via capillary action. This is suitable for small plants, spices or herbs which require relatively low amount of water.

Ebb and Flow systemThe grow beds are flooded with nutrient solution and water using water pump to a required level for some time to provide moisture and nutrients to plants. It is the first commercial system of hydroponics. The major problem in this system is the incident of algae, mould and root rot. A modified structure with filtration unit is made to overcome this problem.

Drip systemThis system can be used for both home garden and commercial units. An appropriate proportion of Water and nutrient solution is supplied to individual plant roots from the reservoir through pump (Rouphael and Colla, 2005). Nutrient solution drips in moderately absorbent growing medium in which plant is grown.

Deep water culture systemThis is simple method where roots of plants are directly suspended in water and nutrient solution and air stone provide air to the roots. The stryofoam makes the platform on which plants float over water. It is suitable for larger fruit bearing plants especially tomato and cucumber. For example, hydroponics buckets system.

Nutrient Film Technique (NFT) systemIn this method circulation of water or a nutrient solution is throughout the entire system and a water pump (without a time control) which pumps solution to the growth tray. The trays are slightly slanted so that nutrient solution after running through roots and goes down back into a reservoir. This system is suitable for various leafy greens production and it is commercially used for lettuce production.

Merits and Demerits of HydroponicsHydroponics is simple method of vegetable production with relatively low cost of cultivation as there are no chances of incidents of soil borne diseases and insect pest and the expenses of fungicides and insecticides are reduced or eliminated. Water consumption is reduced to less than 1/10th - 1/5th of that used in soil cultivation. It is used to grow off season crops as the influence by climate change on plants is minimized. The yield of crop is increased to many folds for example the average yield of lettuce in soil cultivation method is 9-10 ton/acre while in hydroponics its average production is 300-400 ton/acre. Similarly, average production of tomato under soil cultivation and hydroponics system is 10-12 ton/acre and 180-200 ton/acre, respectively. The major demerit that Indian practitioners have to overcome is the high initial cost, requirement of skilled manpower and awareness to the changing market demand.

ReferencesSharma, N., Acharya, S., Kumar, K., Singh, N. and

Chaurasia, O.P. 2018. Hydroponics as an advanced technique for vegetable production: an overview. Journal of Soil and Water Conservation, 17(4): 364-371.

Bulgari, R., Baldi, A., Ferrante, A., and Lenzi, A. 2017. Yield and quality of basil, swiss chard, and rocket microgreens grown in a hydroponic system. New Zealand Journal of Crop and Horticultural Science, 45(2), 119-129.

Son, J.K., Kim, H.K. and Ahn, T.I. 2016. Hydroponic Systems. In: Plant Factory: An Indoor Vertical Farming System for Efficient Quality Food Production, 213-221 https://doi.org/10.1016/B978-0-12-801775-3.00017-2

Rouphael, Y. and Colla, G. 2005. Growth, yield, fruit quality and nutrient uptake of hydroponically cultivated zucchini squash as affected by irrigation systems and growing seasons. Scientia Horticulturae, 105(2): 177-195.

Shrestha, A. and Dunn, B. 2013. Hydroponics. Oklahoma Cooperative Extension Services HLA-6442.



MEDICINAL AND AROMATIC PLANTS

20143

46. Amazing Amla JuiceDR. HARPREET B. SODHI

Assistant Professor, College of Agribusiness Management, S. D. Agricultural University, Sardarkrushinagar – 385506 (Gujarat), India *Corresponding Author Email: [email protected]

IntroductionGooseberry is the berry of species named Ribes which is native of Europe and parts of Asia and Africa. Amla, the Indian gooseberry is a very renowned fruit since many decades for its endeavoring properties and nutritional benefits. With the increasing prominence of medical conditions almost completely taking over people’s lives, it is important to have some amount of an understanding of the various factors that play a role in keeping your body healthy. Besides this, one should also ensure that any complications are minimized to a substantial degree. The ability of the human body is dictated to a significant degree by the kinds of foods that it is provided. However, it is also necessary that one has an understanding of which foods are known to be healthier choices than others as this is likely to put you in a great position to be able to take better care of your body. One of the healthier natural substances that is widely used in a number of home remedy recipes to fight off a variety of medical conditions is the Indian gooseberry. Greek and Indian medicine science advocates that the gooseberry and its juice have a lot of health benefits. Here are some health benefits of Amla Juice.

Health Benefits of Amla JuiceSource of Vitamin C: Some of the more prominent gooseberry benefits revolve around the fact that the tiny berry is extremely high in vitamin C content. Amla juice contains vitamin C that is twenty times more than in orange juice. Vitamin C present in the Amla juice improves the tannins which are required to shield the heat and light. This characteristic makes it the most powerful rejuvenating agent known to man thus far. Because of this vitamin C potency, the gooseberry benefits are taken advantage of by being commonly included in a number 80 of supplements as well as tonics in order to allow the vitamin C to be easily assimilated into the body.

� Brings Natural Glow on Face and Helps Removing Blemishes from Skin: Amla juice works perfectly for the removal of pimples and acne scars from your skin. If you apply the paste made from Amla for 10 to 15 minutes it will heal the spots and decrease the acne affected skin. This way the antiseptic properties of Amla helps skin to look more beautiful and fresher. Drinking Amla juice with the addition of honey in the morning hours

brings natural glow to the face and it also makes the skin free from blemishes.

� Stronger Hair Growth: Applying Amla juice that is mixed with water to the scalp can restore the spirit and energy of the hair. To make hair stronger and shiny, add Amla powder and lemon juice, apply over the scalp and retain it for 20 to 30 minutes and rinse off with warm water. Use can also use the Amla oil for scalp massage. It will strengthen the hair from roots and brings the natural glow to hair. Few people use this to get relief from stress.

� Retains Body Temperature: In the summer season, juice from Amla retains our body cool and sorts out the heat from the body. Amla also works as shield to the radiation and it protects from harmful UV rays, thereby saving our skin from getting exposed to the harsh weather conditions and keeps it hydrated and cool.

� Lowers Osteoclasts: People who suffer from bone and calcium problems should consume Amla as Amla juice lowers the osteoclasts the cells which break down the bones. It also reduces the chronic cough, allergic asthma and tuberculosis.

� Blood Purification: Amla juice when taken with honey everyday twice helps in relieving asthma and bronchitis complications. Amla’s antioxidant properties help the body to purify the unwanted products from your blood and make it healthy. It also works wonder for blood purification.

� Life Span of a Person: Ayurveda states that eating Amla on regular basis or drinking Amla juice increases life span of a human. The fresh Amla comprises of water, fiber, minerals, proteins and carbohydrates which when consumed it make you strong and healthy thereby increasing your life span.

� Urinary Tract Cleanliness: Amla juice also helps to cure Urinary Tract disorder. It also helps to get rid of Urinary Tract infections. Try 30 ml of Amla juice two times daily to get the relief from burning urination.

� Source of Nutrients: Amla also comprises of many minerals and vitamins such as Carotene, Phosphorus, Calcium, Iron, and Vitamin B Complex. Amla is also known to be a powerful antioxidant.

� Increases Red Blood Cells: For people having low count of hemoglobin or red blood cells are



recommended to have Amla juice. Amla juice is responsible to boost up the count of red blood cells in the body.

� Lowers Cholesterol Levels: Studies have listed the gooseberry nutrition water facts as each berry containing about 80 % and the combination of its ingredients helping with substantially lowering cholesterol levels as well. The gooseberry benefits can be taken advantage of in a fresh fruit, juiced or even dry form of the fruit.Other Health Benefit: Amla juice provides

strength to heart muscles, which is a best remedy to treat heart problems. It also prevents the incidence of heart ailments. It strengthens the heart muscles and as such, facilitates the free flow of blood in the body

without any obstruction. It is rich source of iron and helps in promoting the normal functioning of the circulatory and reproductive systems and also act as a body building agent by repairing old cells and forming new ones. Amla juice mixed with turmeric powder with some honey controls diabetes problem. Gooseberry helps in treating various eye ailments including nearsightedness and cataract. Amla juice with honey can be taken to enhance your eye sight. It stimulates bowel movements and helps to cure chronic constipation. Amla juice rejuvenates the body and prevents the signs of aging. It can also be used as an anti-aging remedy. Taking Amla juice also strengthens the immune system, thereby protecting from various infections and diseases.

FORESTRY

20099

47. Calliandra (Calliandra calothyrsus) Fodder: A Protein SupplementCHICHAGHARE AR*

*Department of Silviculture and Agroforestry, College of Forestry, KAU, Thrissur *Corresponding Author Email: [email protected]

Introduction � It is a small, fast growing, multipurpose and N2-

fixing tree originated from Central America � Fresh green or dried leaves are used to feed

livestock. � It is feed as supplement with protein deficient

roughage or grasses � It has moderate palatability to a livestock. � Calliandra trees are vigorous and having good

coppicing ability. � It can be planted as boundary plating or grass-tree

mixture or in agricultural lands � About 3 kg of fresh calliandra fodder replaces 1 kg

of dairy meal without affecting milk yield. � It is having no threat of invasion

Soil and Climate � It thrives well upto 1900 m elevation, bellow

100 cm of annual rainfall and mean temperature above 20°C.

� It can be grown on various types of soils but sensitive to aluminium saturated soil and intolerant to poor drainage and frost also.

� It has ability to withstand dry period of 2-4 months.

Propagation � Propagated by seed � Seeds are harvested dry and dried in shed � Seeds should be soaked for 48 hours before sowing

for obtaining healthy and vigorous seedlings

� Seeds can be sown in polybags or raised beds at 10 X 5 cm spacing

� 3-4-month-old seedlings should be transplanted in pit before or onset of monsoon.

� Double hedge row with 50 cm spacing is very common in boundary planting

� Sometimes one or two rows of callindra intercropped with 4-6 rows of other fodder grasses

Management and Harvesting � Weeding is necessary for first few months due to

slow early growth rate � Watering is necessary for initial year later it

survives by its own. � Basal dose of 2.5 g of P fertilizer can enhance

growth but in case of intercropping no fertilizer required

� Relatively callindra is not infested with any major diseases

� Generally, first cutting carried out after attaining a height of 2 m but in intercropping, it is better to kept height lower to minimize competition.

� Calliandra can sprout very well if we cut even at ground level.

� Cut should be sharp to reduce chances of injuries to plants.

� First cutting should be carried out at 30 cm above ground to produce adequate branches at the base and cutting height can be increased later.

� It can withstand sever pruning. Harvesting interval should be 2 months during the rainy



season and 3-4 months in summer � According to KAU, 60 X 60 spacing with pruning

height of 1 m and harvest interval of 12 weeks gives good yield for Calliandra fodder in humid climate.

� For obtaining optimum fodder in summer, it is recommended to prunned trees 6 months prior.

Nutritive Value � Calliandra fodder can be mixed with napier grass

as supplements to increases crude protein content upto 13% for milch animals.

� Dried leaves can also be fed in summer. � As supplement callindra fed as 1/4 to 1/3 of the

total diet normally. � Calliandra produces 3 kg/m2/year dry matter

from single hedge. � About 250 m long hedge with 50 cm spacing

(500 trees) is required to feed 1 cow a 6 kg fresh calliandra (2 kg DM) per day per year.

TABLE 1: composition and digestibility of calliandra fodder

CP 21 to 25Acid Detergent Fibre 22 to 31%Neutral detergent fibre 45 to 50%Digestibility: 40% to 60%.Ash 4 to 5%Fat 4 to 5%

FIG 1: Two hedge row boundary planting of calliandra fodder in Kerala

FIG 2: Calliandra intercropped with Hybrid Napier grass and with guinea grass

FIG 4: Block planting of calliandra fodder in small holder farm in Kerala

ReferencesAnu Sagaran, K. 2013. Performance of Calliandra

calothyrsus M. under Diverse Management Regimes in a Coconut Based Hedge Row Fodder Production systems. MSc Thesis, Kerala Agricultural University, Thrissur.

Tuwei, P.K., Kang’Ara, J.N.N., Mueller-Harvey, I., Poole, J., Ngugi, F.K. and Stewart, J.L. 2003. Factors affecting biomass production and nutritive value of Calliandra calothyrsus leaf as fodder for ruminants. The Journal of Agricultural Science, 141(1): 113-127.

PLANT BREEDING AND GEENTICS

20134

48. Apomixis In Crop ImprovementTABINDA ALI AND VARTIKA BUDHLAKOTI

Department of Plant Breeding & Genetics, G. B. Pant University of Agriculture & Technology, Pantnagar (Uttarakhand).

Classically, apomixis can be defined as Asexual or agamic reproduction by seeds i.e., Agamospermy. This is in contrast to amphimixis or sexual reproduction (Nogler, 1984). During embryo development there is an

absence of chromosome reduction. Cytological analysis of ovule development process and characterization of genetic variability or uniformity of an individual in comparison to their parents helps deduce the mode of



reproduction of the plant. Apomixis can be of broadly three types viz., adventitious embryony, diplospory (Sporophytic type) and apospory. Sporophytic and gametophytic apomixis differs on the fact that in the former there is no alteration of generations prior to embryo development. The formation of embryo is directly from a somatic cell in the ovule, mostly a nucellar cell and sometimes from the cell of integuments. For seed development nutritious endosperm is of supreme importance and this can be catered to through amphimixis that happens during the gametophytic phase of the lifecycle. Amphimixis is concurrent with Sporophytic apomicts. As a result, reduced embryo sacs and adventitious embryos occur simultaneously. So, after the fertilization of the central cell of reduced embryo sac or polar bodies, endosperm develops that provides nutrition to either an adventitious embryo or zygotic embryo when one out competes the other, or both may develop to relative maturity resulting in poly-embryony.

Obligate Gametophytic apomixis doesn’t have amphimixis. It’s rare to find a completely obligate apomict. They hardly exist. Development of mega-spore mother cell is arrested or it somehow cannot complete meiosis. So, in the absence of meiosis, haploid (n) megaspore cannot develop and all cells of the unfertilized ovule retain somatic chromosome number. When megaspore mother cell or one of its diploid daughter cells develops into an unreduced female gametophyte, this phenomenon is called diplospory. There are various sorts of diplospory viz., mitotic diplospory or Antennaria types which is ubiquitous. In this megaspore mother cell directly undergoes mitosis and another one which occurs in Taraxacum sp. where there is restitution in meiosis I (Nogler, 1984).

In case of Apospory, somatic cell is the origin of unreduced female gametophyte therefore possibility lies that both apomixis and amphimixis occurs in the same ovule in facultative apomicts. This is rare and most of the times one process outcompetes the other. When an unfertilized gamete develops into a plant then the process is called Parthenogenesis. There are some rare cases when an unreduced egg fertilizes resulting in a hybrid embryo with high ploidy levels. There is a syngamy between egg and sperm when meiosis occurs in an apomict. Embryo development from an unreduced egg could be precocious or early e.g., Tripsacum dactyloides. Salmon system in wheat is a good model to explain parthenogenesis in plants. In a particular cytoplasmic background, there is a wheat rye translocation in the nucleus resulting in male sterility and haploid parthenogenesis. Salmon system isn’t apomictic but it provides useful isogenic lines for molecular studies including the construction of egg- cell specific cDNA libraries.

Pseudogamy or fertilization of central cell is really important for the production of endosperm in many apomictic species. There are some exceptions to it e.g., plants of Asteraceae family do not require fertilization and there is an autonomous endosperm development. The maternal to paternal genome contribution to the

endosperm and its relationship to the ploidy level of embryo have a strong effect on seed viability of amphimicts. In maize normal endosperm development happens only when maternal to paternal genome ratio in the endosperm is 2m:1p. (Lin, 1984). In certain panicoid apomicts and in Pennisetum species have four nucleate embryo sacs that is fertilized to create 2n+n endosperms or reconstituting 2m:1p ratio (Ozias-Akins et al., 2003). This change in the developmental pattern occurs when apomixis is introgressed into sexually reproducing Pennisetum glaucum from an apomictic relative. There is an increase in the binucleate central cells as compared to the uninucleate central cells and all this is related to reduced seed set.

Apomixis is widely distributed among angiosperms. Most widespread in the grass (Poaceae) and sunflower (Asteraceae). Apomixis occurs due to deregulation of the sexual processes in various developmental stages. Sporophytic apomixis is a dominant trait and this was studied in a cross Citrus volkameriana X Poncirus trifoliata and 3:1 segregation ratio was obtained in this cross for apomixis. (Garcia et al., 1999). In Apospory when aposporus male parent is crossed with strictly sexual female parent the phenomenon of apospory appears to be simply inherited. Apospory behaves as a dominant trait. The association of apospory with the heterochromatic region of the genome, rich in retrotransposons as in Pennisetum, raises the intriguing possibility that chromatin structure/ RNA Interference could play a role in control of apomictic gene expression.

Gametophytic apomixis has two morphologically different forms viz., diplospory and apospory. They differ in heterochronicity and regulatory control of gene expression. If we want to manipulate the genetic expression of a crop plant then it is imperative to study their genetic mechanism. Crops which are maternally alike are useful for creating hybrids. It remains a conundrum that isolation of the genetic components of apomicts and their transfer to the crop species or synthesis of apomixis from combination of mutations for its components in sexual plants. In addition, interspecific crosses formed as a result of gene escape from apomictic crops into wild relatives will not have meiotic irregularities leading to sterility and therefore will survive under natural conditions (van Dijk and van Damme, 2000). Transfer of apomixis to crop species through wide crosses has not been successful so far, but transgenic technology offers a more powerful way to introgress apomixis into crop species (Kandemir, et al, 2015.). Alteration of a single gene in a sexual plant can bring about functional apomeiosis, a major component of apomixis (Maruthachalam, et al.,2008).

Apomixis is a nuisance when plant breeding is done to produce sexual progeny such as inbreds and hybrids but its of supreme importance when the crosses are to be maintained indefinitely. Plant breeders have developed various schemes to use apomixis for fixing of heterosis and hence the way forward is to avoid apomixis when the crossing is done in the apomictic species but once a hybrid or inbred is created then it can be maintained and multiplied by the process of



apomixis.

FIG.: Various classifications of Apomixis.

ReferencesKandemir, N. and Saygili, I. (2015). Apomixis: New

horizons in Plant Breeding, https://journals.tubitak.gov.tr/agriculture/abstract.htm?id=16663.

Lin B-Y. (1984). Ploidy barrier to endosperm development in maize. Genetics 100:475–486

Maruthachalam, R., Marimathu, P. A. M. and Siddiqi,

I. (2008). Gamete formation without meiosis in Arabidopsis. Nature 451, 1121-1124.

Nogler, G.A. (1984). Gametophytic apomixis. In: Johri BM, ed. Embryology of angiosperms. Berlin, Germany: Springer, 475–51.

van Djik, P., and van Damme, J. (2000). Apomixis technology and the paradox of sex. Trends Plant Sci 5:81-84.

20090

49. RNA Interference: A Novel Approach for Crop ImprovementPAWAN KUMAR1 MANOJ KUMAR2 AND SANDHYA KULHARI3

1Pawan Kumar, Scientist, Indian Institute of Soil and Water conservation, Dehradun-248195, (UK) 2&3Assistant Professor, Agricultural Research Station Ummedganj, Agriculture University, Kota-324001 (Raj.), *Corresponding Author Email: [email protected]

IntroductionGene silencing is a process by which a specific gene is down regulated and probably that evolve as a genetic resistance system against microorganism and invading nucleic acids. There are several routes of gene silencing identified in plants such as: post-transcriptional gene silencing or RNA interference, transcriptional gene silencing, microRNA silencing and virus induced gene silencing. RNA interference (RNAi) term coined by Andrew Fire in 1998 and awarded Nobel Prize for their work on nematode Caenorhabditis elegans in Physiology or Medicine in year 2006. RNA interference (RNAi) is a process that inhibits gene expression by the double-stranded RNA (dsRNA) that can cause the degradation of target messenger RNA (mRNA). It leads to post transcriptional gene silencing (PTGS) activate by double stranded RNA (dsRNA) molecules

to check the expression of particular genes. RNAi is a novel approach to modify the gene expression for better quality traits, nutritional improvement, resistance against pathogen and tolerance to abiotic stress in different crops. Recently, it emerges as a powerful and more determinant technology to study the gene loss of function phenotype which leads to gene functional analysis when no mutant alleles are unavailable.

RNA InterferenceRNAi is an RNA-dependent regulation of gene expression in a cell to prevent the expression of a certain gene process that is controlled by the RNA-induced silencing complex (RISC). It is initiated by short double-stranded RNA molecules in a cells cytoplasm, where they interact with the catalytic RISC component containing argonaut that is protein family



and plays a central role in RNA silencing processes. When the dsRNA catalyzed by RISC complex, the RNA is imported directly into the cytoplasm and cleaved to short fragments by Dicer. The RNAi involve two premier pathways are RNA introduction and mRNA degradation. RNA interference pathway mostly follows four common steps such as (i) dsRNA cleaved by dicer (ii) entry of SiRNA into RISC complex (iii) silencing complex activation (iv) mRNA degradation. In the first step of RNAi, the dsRNA in the cell, which is perfectly homologous in sequence to the target gene, is recognise by DICER enzyme and siRNA of 21 - 25 nucleotides produced. Then, the SiRNA incorporated into multi-component nucleus complex into the RNA induced silencing complex (RISC). The next step involves unwinding of the SiRNA duplex by helicase enzyme and further re-modelling of the complex to create an active form of the RISC. The active component of an RISC is endonuclease called agronaute protein which cleave the target mRNA strand complementary to their bound SiRNA and degrade the mRNA. After the cleavage is complete, the RISC departs and the SiRNA can be reused in a new cycle of mRNA recognition and cleavage. Through the cleavage of dsRNA into small SiRNA by dicer result in some degree of amplification, it is not sufficient to bring about continuous mRNA degradation. It provides very convincing genetic and biochemical evidence that RNA dependent RNA polymerase plays a vital role in increasing the RNAi effect. So, RNAi technology is coming out as a convincing approach in which short dsRNA

suppress the expression of specific gene by inducing the homologous sequence of the target mRNA in the cytoplasm.

Application of RNAi for Nutritional Value in Crop PlantRNAi process holds the key to future technological applications. The ultimate goal of genome sequencing projects to accelerate the identification of the biological function of genes. RNAi technology is proving to be useful to analyze quickly the functions of a number of genes in a wide variety of organisms. Given the gene-specific features of RNAi, it is conceivable that it will play an important role in crop improvement in many ways. It is also used therapeutic applications and plant anti-nutritional factor could also be removed if the anti-nutritional biosynthesis genes are targeted with the RNAi constructs.

Biotic and Abiotic Stress ResistanceThe RNAi strategies could be employed to control the biotic and abiotic stresses by improving defense mechanism in crop plants. However, further research is needed to explore the novel strategies amounting to efficient and durable gene silencing. Efficacy of RNAi was suggested by many researchers for better control over the parasitic weeds, virus, bacteria, fungi, nematodes and insects (Table 1). The miRNA plays important regulatory roles in development and stress response in plants by negatively affecting post-transcriptional gene expression.

TABLE 1 The gene targeted for crop improvement through RNAi technology

Trait Target Gene Host Application ReferenceEnhanced nutrient content

Lyc Tomato Increased concentration of lycopene (carotenoid antioxidant)

Sun et al. (2012)

APX Tomato Higher Vitamin C content Zhang et al. (2011)SBEII Wheat, Sweet

potato, MaizeIncreased levels of amylose for glycemic management and digestive health

Regina et al. (2006)

DET1 Brassica napus Carotenoid with decreased sinapate esters

Wei et al. (2009)

SBEII Barley Amylose content Regina et al. (2010)FAD2 Canola, Peanut,

CottonIncreased oleic acid content Liu et al. (2002)

SAD1 Cotton Increased stearic acid contentZLKR/SDH Maize Lysine-fortified maize Houmard et al. (2007)

Reduced alkaloid production

CaMXMT1 Coffee Decaffeinated coffee Ogita et al. (2003)COR Opium poppy Production of non-narcotic

alkaloid, instead of morphineAllen et al. (2004)

Altered phenotype TA29 Tobacco Male sterility Nawaz-ul-Rehman et al. (2007)

BCP1 Arabidopsis thaliana Male sterility Tehseen et al. (2010)orfH522 (male sterile)

Tobacco Fertility restored Nizampatnam and Kumar (2011)

Ethylene sensitivity

LeETR4 Tomato Early ripening tomatoes Meli et al. (2010)ACC oxidase gene

Tomato Longer shelf life because of slow ripening

Xiong et al. (2005)



Trait Target Gene Host Application ReferenceReduced production of lachrymatory factor synthase

lachrymatory factor synthase gene

Onion “Tearless” onion Eady et al. (2008)

Gene targeted for biotic stress resistance through RNAi technology

S. no.

Defence improvement Resistance against Targeted gene Plant used

1 Insect resistance Helicoverpa armigera CYPAE14 CottonCorn rootworm V-ATPase A Maize cyst nematodes miR396 Arabidopsis

2 Virus resistance Rice Dwarf Virus (RDV) PNS12 RiceBean golden mosaic virus (BGMV) AC1 gene BeanRice Stripe Virus CP gene and disease-specific protein Rice

3 Nematode resistance

Meloidogyne incognita splicing factor and integrase TobaccoMeloidogyne 16D10 Arabidopsis

4 Bacteria resistance

Xanthomonas citri subsp. citri (Xcc) PDS and CalS1 Lemoncrown gall tumours (Agrobacterium tumefaciens)

(iaaM and ipt) Arabidopsis

5 Fungus resistance

Phytophthora infestans SYR1 Potato

(Modified from Saurabh et al., 2014)

Extensive studies of miRNAs have been performed in model plants such as rice, A. thaliana and other plants. Wang et al. (2011) identified drought-responsive miRNAs in a legume model plant, Medicago truncatula. They reported that 22 members of 4 miRNA families were upregulated and 10 members of 6 miRNA families were down-regulated in response to drought stress. The Arabidopsis seedlings were exposed to different abiotic stresses such as drought, cold, salinity, and oxidative stress. Chen et al. (2012) provided valuable information of miRNAs in Al3+ toxicity and tolerance. Sunkar and Zhu (2004) described that miR393 was strongly up-regulated by cold, dehydration, high salinity and abscisic acid (ABA) treatments. Several stress-related miRNAs have been discovered in rice and only two miR393 and miR169g have been found to be related to abiotic stress; both were up-regulated by dehydration (Zhao et al. 2007). Hwang et al. (2011) identified drought stress-responsive miR171 family members (miR171a, miR171b and miR171c) in potato (Solanum tuberosum) plants.

Future ProspectsThe global demand for food is likely to increase with the continuing population growth and consumption. Malnutrition is a major problem especially in developing countries. To ensure a healthy diet for healthy world, there is a necessity to develop bio-fortified cereals, fruits and vegetables with nutritionally enrichment. On other hand crop plants should be developed for the stress (biotic and abiotic) tolerance. So, there is a need to develop plant varieties that will tolerate phytopathogens and pests, along with changing environmental conditions. RNAi based researches have proved its potential in crop improvement to overcome the problem of food security, malnutrition and famine. With RNAi, it would be possible to target

multiple genes for silencing using a thoroughly-designed single transformation construct. Moreover, RNAi can also provide broad-spectrum resistance against pathogens with high degree of variability, like viruses. The crop plants developed through RNAi should undergo risk assessment related to food safety and environment protection. So, there is a need to tailor customize vectors according to the necessity of crop improvement. Although much progress has been made on the field of RNAi over the past few years, the full potential of RNAi for crop improvement remains to be realized.

ReferenceSaurabh S., Vidyarthi A.S. and Prasad D. 2014 RNA

interference: concept to reality in crop improvement. Planta, 239: 543–564. doi: 10.1007/s00425-013-2019-5.

Wang T., Chen L., Zhao M., Tian Q., Zhang W.H. 2011 Identification of drought-responsive microRNAs in Medicago truncatula by genome-wide high-throughput sequencing. BMC Genomics,12: 367. doi: 10.1186/1471-2164-12-367.

Chen L., Wang T.Z., Zhao M.G., Tian Q.Y. and Zhang W.H. 2012 Identification of aluminum-responsive microRNAs in Medicago truncatula by genome-wide high-throughput sequencing. Planta, 235: 375–386.

Sunkar R, Zhu JK. 2004 Novel and stress-regulated microRNAs and other small RNAs from Arabidopsis. Plant Cell, 16:2001–2019.

Zhao B., Liang R., Ge L., Li W., Xiao H., Lin H., Ruan K. and Jin Y. 2007 Identification of drought-induced microRNAs in rice. Biochemical and Biophysical Research Communications, 354: 585–590. doi: 10.1016/j.bbrc.2007.01.022.

Hwang E.W., Shin S.J., Yu B.K., Byun M.O. and Kwon H.B. 2012 miR171 Family Members are involved in



Drought Response in Solanum tuberosum. J Plant Biol, 54: 43–48, doi 10.1007/s12374-010-9141-8.

20116

50. Model Organisms for Genetic StudiesPRASANTA K. MAJHI1, MOUNIKA KORADA2 AND SONALI V. HABDE3

Research Scholar, Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, U.P., India *Corresponding Author Email: [email protected]

Model OrganismsThe model organisms are used in the biological research because i) they may help overcomes ethical and experimental constraints that hold for the target life form, ii) they provide a framework on which to develop and optimize analytical methods that facilitate and standardize analysis, and iii) they are thought to be representative of a larger class of living beings for whatever biological phenomenon or process the community is interested in. Therefore, it is necessary

to predict how similar different organisms might be with respect to biological processes of interest. The availability of fully sequenced genomes of the model organisms and the advances in comparative genomics and computational systems biology allows us to understand the genetic networks for exploiting the behavior of specific processes between the organisms. Here, some of the organisms are discussed along with their desirable features which make them as model organisms for biological, physiological, molecular and genetic studies.

Model organisms Desirable Features Arabidopsis thaliana (Wild Mustard, 2n=10)Family: Brassicaceae

Small genome size (114.5 Mb/125 Mb total) has been sequenced in the year 2000 (Sequence Viewer, AGI).A rapid life cycle (about 6 weeks from germination to mature).

Escherichia coli (E. coli)[Gram negative, facultative, anaerobic, rod-shaped and coliform bacterium]

The phenomenon bacterial conjugation was first studied by Joshua Lederberg and Edward Tatum (1946) using E. coli as a model bacterium.It is the first organisms to have its complete genome sequence of E. coli K12 was published by Science in 1997.The work of Stanley Norman Cohen and Herbert Boyer in E. coli, using plasmids and restriction enzymes to create recombinant DNA, became a foundation of biotechnology. From 2002 to 2010, a team at the Hungarian Academy of Science created a strain of Escherichia coli called MDS42, which is now sold by Scarab Genomics of Madison, WI under the name of “Clean Genome”. One of the first useful applications of recombinant DNA technology was the manipulation of E. coli to produce human insulin.Major attributes that make E. coli an excellent model organism:Genetic Simplicity: E. coli cells only have about 4,400 genes whereas the human genome project has determined that humans contain approximately 30,000 genes.Single-celled organism: There are no ethical concerns about growing, manipulating, and killing bacterial cells, unlike multicellular model organisms.Rapid growth and reproduction: Double its population about every 20 minutes.Foreign DNA Hosting: The genetics of E. coli are well under stood, and can be readily manipulated or engineered.

Saccharomyces cerevisae (Baker’s or Budding yeast):

Up to 30% of genes implicated in human disease may have orthologs in the proteome of yeast.Genome size of ~12 Mbp with full genome sequence in 1996 as the first eukaryotic organism to be sequenced including protein-DNA, protein-protein, and genetic interactions.Research area: Aging, regulation of gene expression, signal transduction, cell cycle, apoptosis, programmed cell death, protein folding.In recent, the successful identification of a highly conserved regulatory complex implicated in human leukemia complex, named COMPASS (Complex of Proteins Associated with Set1), was originally identified by studying protein interactions of the yeast Set1 protein, which is the ortholog of the human mixed-lineage leukemia (MLL) gene.

Caenorhabditis Elegans (Nematode)

Short lifespan i.e. 2-week, easy for culture, rearing and feed.Research area: Ageing processes, genetically and physiologically disorders.Used to study neural development; due to the availability of a comprehensive connectivity map and only 302 neurons and ~7000 synapses.



Model organisms Desirable Features Danio rerio (Zebrafish) Approximately, 80% of the genes involved in human diseases are equivalent in Zebrafish.

Very well characterized genome size ~1.5 Gbp.Most of the organ systems in Zebrafish are like humans, for example, heart, eyes and kidneys, therefore, organ development can be studied.

Mus musculus (Mouse, 2n=40)

The genome has 85% similarity with human.Genome size ~2.5 Gbp.Research area: Ageing, high blood pressure obesity, heart disease, neurodegenerative disorders and stress.

Rattus norvegicus (Rat) The first genetic studies were carried out by Crampe from 1877 to 1885 and focused on the inheritance of coat color.Research Area: Autoimmune diseases, aging, breast cancer, blood diseases, eye disorders, cardiovascular diseases, neuromuscular diseases and kidney diseases.

ReferencesKlaudia, M. and Wojciech, P., (2020). The Natural

History of Model Organisms: The Norway rat, from an obnoxious pest to a laboratory pet. Polish Academy of Sciences, Poland, DOI: 10.7554/eLife.50651.

Online Source: https://andor.oxinst.com.

Phillips, T. (2019). E. coli is Critical to Genetic Advances. https://www.thoughtco. Com.

Shahin, M., Baharak, S., Shankar, S. and Ananth, G. (2015). Scope and limitations of yeast as a model organism for studying human tissue-specific pathways. BMC Systems Biology, 9:96. DOI 10.1186/s12918-015-0253-0.

20120

51. Molecular Cross-Talk between Ethylene and Erfs for Submergence Tolerance in RiceKORADA MOUNIKA*, PRASANTA KUMAR MAJHI AND RASHMI K

Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, U.P., India *Corresponding Author Email: [email protected]

IntroductionEthylene, the first identified simplest gaseous phyto-hormone, has a role in regulating diverse physiological pathways in plants. There is a significant increase in the biosynthesis of ethylene under different stress conditions as well as during various developmental stages such as seed germination, root elongation, tissue differentiation, organ abscission, leaf and flower senescence (Schaller and Voesenek, 2015). Likewise, in rice, the genetical, physiological and morphological processes are regulated by ethylene under various stress conditions.

Transcription factors (TFs) are proteins that help turn specific genes “on” or “off” by binding to nearby DNA and regulate a number of biological processes under normal and stress conditions. APETALA2(AP2)/ethylene response factor (ERF) is a transcription factor which plays a major role in stress response in rice. Ethylene signaling cascade involves a variety of transcription factor families in rice. The regulation of stress-responsive genes by transcriptional regulatory factors, also known as transcriptome reprogramming, helps in rice under critical environmental stresses. The up- or down- regulation of ethylene signaling pathway under adverse stress conditions results in expression

of many stress related genes in various families and among them AP2/ERF superfamily is one of the largest TF super families in crop species.

Molecular Cross-talk between ERFs and ethylene is for Submergence Tolerance in Rice. The synergistic or antagonistic functions of different Plant Growth Regulators indicate that ethylene plays a major role in the acclamatory responses to flooding and submergence in rice. Under conditions of low oxygen, which occurs during submergence, ethylene helps in regulating the cell death, leading to formation of adventitious roots and aerenchyma, and induces petiole and internode elongation. As a result, the photosynthetic organs of rice extend above the water surface. The process of diffusion of ethylene is slower in water as compared to air and due to this ethylene concentration rises rapidly and stimulates initiation of coleoptiles, mesocotyl, leaf, internode and petiole elongation under submerged conditions of rice (Kendrick and Chang, 2008).

The quiescence mechanism is a vital concept to induce submergence tolerance in rice, where elongation of the plant is inhibited during flooding and there will be subsequent re-growth when the water recedes. Recent molecular and genomic analysis have identified a quantitative trait locus (QTL) designated



as Submergence1 (Sub1), encoding a cluster of up to three ethylene-responsive factor (ERF) genes- Sub1A, Sub1B and Sub1C. Sub1A primarily promotes submergence tolerance in lowland rice whereas Sub1B and Sub1C are regulatory factors that are induced in a wide variety of Japonica and Indica rice cultivars. All of these Sub1 genes belong to the B-2 subclass of

ethylene response factors (ERFs)/ which possess a single 58- to 59-residue ERF domain. Some genes like the SNORKEL1 and 2 (SK1 and SK2) are additional Sub1A-related genes in deep water rice that play a role in ethylene-induced internode elongation during low oxygen conditions (hypoxia), excess water (flooding) and submergence (Hattori et al., 2009).

FIG.: Diagram representing the link between ethylene and gene expression under submergence in rice. The activity of Sub1A gene is observed in the ethylene signalling cascade under submergence stress in rice. (Perata and Voesenek, 2007)

A detailed study of the Sub locus has shown two alleles of Sub 1A, namely Sub1A-1 (the tolerance-specific allele) and Sub1A-2 (intolerance-specific allele). When Sub1A-1 allele is introgressed into submergence-intolerant rice variety, Sub1C expression has been down-regulated in addition to the up-regulation of alcohol dehydrogenase 1 (Adh1) expression (Fig.). The Sub1 locus containing Sub1A-1 allele indicates negative regulation of ethylene and GA for carbohydrate consumption and cell elongation and positive regulation for ethanolic fermentation during submergence (Abiri et al., 2017). The obtained results indicate that the overexpression Sub1A-1 enhances the submergence tolerance in rice crop.

References1. Schaller, G.E., Voesenek, L.A. (2015). Focus on

ethylene. Plant Physiol. 169: 1-22. Kendrick, M.D., Chang, C. (2008). Ethylene

signaling: new levels of complexity and regulation. Curr. Opin. Plant Biol. 11(5): 479-485.

3. Abiri, R., Shaharuddin, N.A., Mahmood, M., Yusof, Z.N., Atabaki, N., Sahebi, M., Valdiani, A., Kalhori, N., Parisa, A., Hanafi, M.M. (2017). Role of ethylene and the APETALA 2/ethylene response factor superfamily in rice under various abiotic and biotic stress conditions. Environmental and Experimental Botany. 134: 33-44.

4. Hattori, Y., Nagai, K., Furukawa, S., Song, X.-J., Kawano, R., Sakakibara, H., Wu, J., Matsumoto, T., Yoshimura, A., Kitano, H. (2009). The ethylene response factors SNORKEL1 and SNORKEL2 allow rice to adapt to deep water. Nature. 460(7258): 1026- 1030.

5. Perata, P., Voesenek, L.A. (2007). Submergence tolerance in rice requires Sub1A, an ethylene-response-factor-like gene. Trends Plant Sci. 12 (2): 43-46.

20125

52. Crop Wild Relatives of Maize, Rice and Cotton for Tolerance to Abiotic StressBASAVARAJ P S

Scientist Plant Breeding, ICAR-National Institute of Abiotic Stress Management-Pune

Demand for food is estimated to approximately twofold by 2050 because of explosion of population growth of world. In the past “Green Revolution: significantly contributed world food production and food security,

however, restricted availability and constant reduction of various resources such as cultivable land and irrigation resources are severe limitation for future food security of the world the future years. Development of



high yielding cultivars with resistance to abiotic and abiotic stresses, enhanced nutritional quality and the capacity to acclimatize to the ever-changing climate are necessary for sustainable agriculture. Limited genetic diversity of present-day varieties/hybrids is becoming a foremost setback for crop improvement programs. Hence, exploiting crop wild relatives (CWRs) is a favorable approach to broaden genetic base of modern elite varieties. Studies on characterization of genetic resources and breeding programs in the past have demonstrated that modern high yielding varieties/hybrids have lower level of tolerance to biotic and abiotic stress in comparison to crop wild relatives.

Crop Wild Relatives (CWR) can be defined as wild plant species that are genetically related to crops, but have not been domesticated. CWRs provide a diverse array of traits with the potential to reduce the extent of yield loss as a consequence of stresses. These CWR- derived resistant traits transferred to susceptible modern crops through conventional breeding (if there is a sexual compatibility), transgenesis, or other recent techniques. Linkage drag is potential bottleneck associated with introgression of traits through conventional methods arising from CWR, conversely, marker-assisted backcrossing breeding drastically eliminates with least number of generations. Transgenes is a preferred over other methods, if biological constraints posing barrier to transfer desirable loci from CWRs to crops. Theoretically, transgenesis is an attractive due to no linkage drag, however, practically; it is a time taking and costlier process. Transgenesis also depends on the complexity of the donor genome, availability of an efficient transformation system for the recipient crop, and, last but not least, the final product will have to undergo a lengthy, expensive, and complicated deregulation process.

Maize (Zea mays): is a principal crop of the world predominantly grown for animal feed and biofuel. Maize is affected by many abiotic stresses like drought, acidic soils, waterlogging and mineral toxicity etc. causing severe yield losses. Many of the wild relatives known to provide resistance to these stresses are provided in the table.

TABLE 1. Maize crop wild relatives for abiotic stress tolerance

Abiotic Stress

Crop wild relatives

Putative cause of resistance/tolerance

Drought tolerance

Eastern gamagrass

Deep root system

Acid soil Eastern gamagrass

Unknown

Aluminum tolerance

Eastern gamagrass

Unknown

Salinity tolerance

Eastern gamagrass

Conserve sodium in the leaves; Maintaining the turgor pressure; Mechanism of highly efficient sodium ion release from the tissue

Abiotic Stress

Crop wild relatives


Waterlogging tolerance

Zea nicaraguensis

Barrier to radial oxygen loss in basal areas of adventitious roots

Z. luxurians UnknownEastern gamagrass

Constitutive formation of root aerenchyma

Rice (Oryza sativa): second principal food crop of the world contributing to the majority of dietary intake for nearly half of the global population, but known to suffer from many of abiotic stresses. Among various stresses drought, heat, cold, acidic soil, salinity and aluminum toxicity are of great significance. Genetic variability for resistance/tolerance to these stresses is limited in the germplasm of cultivated rice, however, wild species of Oryza are considered rich sources of unexplored genes for these traits. Identification and transfer of these genes from wild species to modern cultivars is, therefore, an attractive option and details of the study are given in table 2.

TABLE 2. Crop wild relatives of Rice against abiotic stress tolerance

Abiotic Stress Crop wild relatives

Resistance/tolerance genes, gene loci, and QTL identified in wild species

Drought and Heat tolerance

Oryza glaberrimaO. barthiiO. meridionalisO. australiensisO. longistaminata

Several gene/QTL identified in O. meridionalis

Acid soil and aluminum tolerance

O. rufipogon Several QTL identified in O. rufipogon

Salinity tolerance

O. coarctata Genomics and transcriptomics studies to underway to identify key genes/pathways

Cold tolerance O. rufipogon QTLs identified in O. rufipogon

Cotton (Gossypium Spp): Globally cotton is primarily grown as fiber and oil seed crop. Genus Gossypium constitute of about 50 species, including five allotetraploids, and other diploid species. Drought, heat and salinity are the major constraint in cotton production. Many wild relatives of cotton significantly associated with tolerance to several abiotic stresses, exploitation of these through pre-breeding or introgression breeding is one of the promised approaches to incorporate tolerance/resistance in cotton. Important species of cotton are provided in table 3.



TABLE 3. Crop wild relatives of cotton against abiotic stress tolerance

Abiotic Stress Crop wild relatives


Drought tolerance G. tomentosumG. herbaceumG. darwinii

Unknown

Salt tolerance G. tomentosumG. davidsoniiG. aridum

Unknown

Heat tolerance G. tomentosum Unknown

Conclusion: Various abiotic stresses, climate vagaries, and rapidly growing demand for food production are causing serious threat to world agriculture. Development of superior cultivars by exploiting diverse sources of genetic variation and recent molecular and genomic tools has the potential to improve food production process. CWRs serve as

valuable resources for genetic improvement of crop plants. Leveraging the untapped genetic diversity available in CWRs for improvement of crops is an attractive option for improving crops. The use of modern technologies for identifying and dissecting the molecular, genetic, and genomic bases of traits in CWRs can accelerate this process in the line of abiotic stress tolerance.

ReferencesMammadov J, Buyyarapu R, Guttikonda SK,

Parliament K, Abdurakhmonov IY and Kumpatla SP (2018) Wild Relatives of Maize, Rice, Cotton, and Soybean: Treasure Troves for Tolerance to Biotic and Abiotic Stresses. Front. Plant Sci. 9:886.

Mondal TK, Henry RJ (2018) The Wild Oryza Genomes. Compendium of Plant Genomes (Chittaranjan Kole edition), Springer Publishers. ISBN 978-3-319-71997-9

20144

53. Transgenic Male Sterility in Crop Plants: A ReviewDR. G. B. SAWANT1*, DR. G. R. GOPAL2 AND DR. S. G. MORE3

1&2Assistant Professor, Department of Agricultural Botany; 3Assistant Professor, Department of Horticulture, Aditya Agriculture College, Beed- 411 122 (Maharashtra) *Corresponding Author Email: [email protected]

Male sterility, as a pollination control tool, gains the utmost importance in development of any hybrid and thus genetic improvement of any crop. It is nothing but the failure of plants to produce functional anthers, pollen or male gametes (Rick, 1944). Koelreuter was the first to report this male sterility in flowering plants by observing anther abortion in some plant species and some inter-specific hybrids (Darwin, 1876). Whereas the first manually developed male sterility system (Cytoplasmic-Genetic Male Sterility) was reported by Jones in onion in 1943. Subsequently the reports of using male sterility systems in many crops as in carrot by Welch and Grimball (1947), induced ‘cytoplasmic male sterility’ in pearl millet by Ethidium bromide (Burton and Hanna, 1976) have become the cases of some of the historical achievements. This male sterility can arise due to different causes as described by Kaul (1988). Manifestation of male sterility traits is associated with morphological changes (complete absence of anthers, reduced anthers), histological changes (premature breakdown of tapetum, delayed collase activity, decreased esterase, reduced growth regulators), irregular micro sporogenesis, biochemical changes (decreased amino acid content, proteins, enzymes), decreased ATP production, etc. Male sterility could be classified into 5 major classes viz. 1) Genetic male sterility (GMS) including both Environment insensitive and Environment sensitive genic male sterility; 2) Cytoplasmic male sterility (CMS); 3) Cytoplasmic-genetic male sterility (CGMS);

4) Transgenic genetic male sterility and 5) Chemically induced male sterility which is non-genetic in nature and thus non heritable.

Transgenic Genetic Male Sterility: Need and CommencementThe conventional approaches of Genetic and cytoplasmic-genetic male sterility suffer through a number of constraints as absence of marker genes in GMS that does not permit the sorting of male sterile and fertile progeny, undesirable effect of the cytoplasm on progeny phenotype, non-availability of suitable restorer gene sources or if available, their unsatisfactory fertility restoration may be due to unsatisfactory pollination, etc. Further environmental effect may cause breakdown of male sterility as apparent especially in environment-sensitive male sterility. These and other reasons confine the applicability of these systems to the limited crops. Transgenic male sterility on the other side provides a novel approach rendering the possibility of overcoming these constraints employing the gene cloning strategies.

It is a type of genic male sterility that incorporates the Transgene i.e. a gene introduced into genome of an organism from any related or unrelated organism by genetic engineering that induces male sterility in that plant. The transgene construct contains the male sterility gene within an expression cassette of appropriate site-specific promotors, operators, regulators along with desired marker genes that



facilitate sorting of individual transgenic and non- transgenic and thereby male sterile or non-sterile plants. Such genetically modified male sterile plants developed after genetic transformation are known as ‘transgenic male steriles’. The first transgenic genetic male sterile was devised by Mariani et al. (1992) in tobacco using barnase/barstar system, the only one system currently in commercial use by which the SeedLink Invigor firstly successfully introduced a GE canola hybrid in 1996 (Department of Agronomy, Iowa State University) with low levels of erucic acid (below 2%) and glucosinolates (30 µmoles/g (CODEX 1999), and are often referred to as “double low” varieties.

Many approaches to develop transgenic male steriles manipulate any of the male sterility manifestation mechanisms given above that disrupts male fertility at different stages. But the next step of fertility restoration is also essential to be searched for the commercial use of these mechanisms in hybrid seed production.

Approaches for Development of Transgenic Male SterilityA. Cell Cytotoxicity

1) Causing pollen abortionThe first transgenic male sterility system devised by Mariani et al. (1992) in tobacco employed this mechanism of pollen abortion using a chimeric gene construct consisting of a pollen or anther tapetum-specific promoter, a cytotoxic gene and a transcription terminator to transform normal male fertile to male sterile plants. They fused TA29, the tapetum-specific promoter isolated from tobacco anthers, with barnase gene from Bacillus amyloliquefaciens and with RNase T1 gene from Aspergillus oryzae to be introduced into tobacco and oilseed rape through genetic transformation, respectively. These RNAses selectively degenerate only tapetal layer under TA29 promotor and prevent pollen formation leading to male sterility. Both ribonuclease genes were closely linked to the bar gene which confers resistance to the herbicide Phosphinothricine (PPT) spraying of which eliminates male fertile, PPT- susceptible plants in the field assuring 100% F

1 hybrid seeds on the male-sterile

plants. Barstar is another gene from same Bacillus amyloliquefaciens which in dominant form encodes a protein inhibiting the Barnase RNAse and thus restores the male fertility. It can be utilized either by inserting it in the same expression cassette formerly producing male sterility or by transforming separate plants with it and their onward use in hybrid seed production. Since barnase male sterility gene is dominant, the male sterile plants (Barnase/-) can be maintained by crossing to any normal male fertile line (-/-). Later using this scheme, some laboratories successfully reported the transgenic male steriles development in oilseed rape, wheat, rice, maize, cotton and also in vegetables viz. cauliflower, tomato, cabbage, lettuce, watermelon and eggplant.

2) Genetic manipulation of hormone levelsSome reports have demonstrated the induction of male sterility by changing endogenous hormone levels to the limits causing cell cytotoxicity. Spena et al. (1987) reported two such genes inducing male sterility in tobacco viz. “rol b” and “rol c” from Agrobacterium rhizogenes driven by the tissue specific promoter ‘tap l’ and ‘35S CaMV’, respectively, flanked with herbicide resistance gene that greatly impaired the flower development due to increased indole acetic acid and decreased gibberellin levels in anthers. Restoration of fertility can be achieved through transformation of other cultivars with the antisense gene inhibiting the expression of male sterility gene (Schmulling et al., 1993). Huang et al. (2011) have acquired a patent on methods of creating reversible transgenic male sterility using tissue-specific promoters expressing cytokinin oxidase gene ckx1 inhibiting pollen formation or male organ development in plants which could be reversed by simple application of cytokinin, a cytokinin oxidase inhibitor or any combination thereof.

3) Inducible male sterility by foreign proteinsA patent is awarded to Albertsen et al. (1999) for method of creating male sterility by anther-preffered expression of high levels of avidin or streptavidin which is a tetrameric biotin- binding glycoprotein derived from both avians and amphibians, found in egg white and also having insecticidal properties. Restoration of fertility can be achieved simply by timely application of biotin, a cofactor that plays a role in multiple eukaryotic biological processes and towards which avidin has a strong and specific affinity. They produced transgenic plants using this system in corn, soybean, canola and sunflower.

4) Pollen self-destructive engineered male sterilityMc Cormick et al. (1989) and Wood (1990) demonstrated that high endogenous levels of auxins like IAA can be combined with pollen self-destructive mechanisms. They used a chimeric gene consisting of pollen-specific promoter ‘LAT59’ and a gene ‘fins2’ that selectively converts indole acetamide (IAM) into IAA at very high concentrations destroying the pollens and render the plants male sterile. Although the detailed characterization of such a transgenic was not reported, this technology would eliminate the tedious maintenance of male steriles and fertility restoration in F

1 hybrids. A single spray of IAM would

compensate the necessity of spraying gametocides/CHAs repeatedly at a specific growth stage and interval. Another such chimeric combination inducing male sterility involves TA-29 promoter with a gene encoding β-glucuronidase (GUS) enzyme which after spraying with protoxins like sulfonyl urea or maleic hydrazide, cause male sterility through their breakdown in the tapetum. The similar inducible destruction of tapetal cells was presented by Kriete et al. (1996) in tobacco and rapeseed by TA-29 promotor driven expression of the gene ‘argE’ encoding L-ornithinase enzyme



that deacetylates the non-toxic herbicide derivative N-acetyl-phosphinothricin leading to the release of the active herbicide and thereby to only tapetal cell death in all the transgenic plants.

One more system, the RHS system also known as glyphosate-mediated male sterility system developed by Monsanto (Feng et al., 2014) in maize also uses such engineered pollen destructive mechanism produced by RHS and RR transgene constructs of which the RHS transgene cassette containing the CP4-EPSPS gene encoding 5-enolpyruvyl-shikimate 3-phosphate synthase impart resistance in plant body to the herbicide glyphosate except in tapetal cells and microspores due to its poor expression there when driven by the enhanced 35S promoter. The RR transgene construct comprises a double expression cassette providing high constitutive expression of CP4-EPSPS which imparts robust resistance to glyphosate in whole plant body keeping male fertility intact and thus enables the hybrid seed production by simply over-the-top sprays of glyphosate in fields having interplanted rows of an RHS female line with those of an RR male line.

B. Male Sterility by Antisense RNA or RNA Interference (RNAi) TechnologyMale sterility in plants may be produced by Antisense RNA and RNA interference (RNAi) technique in which introduction of double stranded RNA (dsRNA) into a diverse range of organisms and cell types causes transcriptional and post-transcriptional silencing of target genes, here in the case, the genes involved at different stages of pollen development. The research group of Cao found some differentially expressed fragments between the genic male sterility line and the maintainer line of Chinese cabbage-pakchoi through mRNA differential display and cDNA-AFLP analysis, and obtained the full-length of genes using RACE technique and verified their function using antisense RNA and RNAi. They transferred a number of antisense genes inhibiting the pollen development and pollen germination in Chinese cabbage-pak-choi and broccoli, e.g. anti-gene CYP86MF encoding cytochrome P450 (Yu et al., 2004; Huang et al., 2005), anti-gene BcMF3 and BcMF4 (Liu et al., 2006; 2007) and antisense DAD1 encoding phospholipase A1 (Chen et al., 2009; 2010). The antisense actin gene was transferred into tomato giving rise the plants with abnormal and nonviable pollen cells. But on pollination with wild type plant pollen, these transgenics produced normal seeds ensuring the fertility restoration, steady inheritance and no any other morphological differences.

C. Male Sterility through Early Degradation of CalloseCallose wall degradation by callose enzymes such as β-1,3-dextranase or glucanase is a key and strictly time-limiting process in normal pollen development and it takes place only after microspore has formed the exine. These callose enzymes are secreted by tapetum cell layer liberating the microspores into anther cells.

Curtis et al. (1996) transformed lettuce plants with gene encoding β-1,3- dextranase linked to tapetum-specific promoter, causing early degradation of developing microspore callose wall and leading to male sterility. Similarly, Scott et al. (2008) have produced male sterile transgenic tomato plants by fusing glucanase gene with tapetum-specific promoter A3 or A9.

D. Male Sterility through Modification of Biochemical Pathways

1) FlavonoidsFlavonoids are one of the major flowers colouring pigments in plants which also have role in reproduction and defence-related mechanisms. A large number of genes are involved in the biochemical pathway of flavonoids synthesis which are produced as phenyl-propanoid based secondary metabolites via chalcone synthase (Chs) (Forkmann, 1991). The gene encoding chalcone synthase (Chs) under control of CaMV 35S promoter has tapetum-specific expression and is directed by anther box as evident from the reduction of anther and pollen pigmentation in about 14 % of transgenic plants. The copy number and orientation of the inserted anther box did not affect the antisense phenotype in a quantitative or a qualitative way.

2) Jasmonic acidJasmonic acid (JA) plays an important role in regulating flower opening, anther dehiscence and pollen maturation and is synthesized from linolenic acid (LA) through octadecanoid pathway (Weiler, 1997). Ishiguro et al. (2001) identified an Arabidopsis mutant defective in anther dehiscence (dad1) showing impaired anther dehiscence due to the reduced accumulation of JA in flower buds whereas the counterpart DAD1 was found to encode a chloroplastic phospholipase A1 to supply free LA catalysing the initial step of JA biosynthesis. Exogenous application of JA or LA rescued the impaired dehiscence making the plant again male fertile.

3) CarbohydratesCarbohydrates play essential role in sustaining growth and signals that influence pollen and anther development in vivo and in vitro (Pacini, 1996; Clement and Audran, 1999). During carbohydrates supply, extracellular invertase mediates phloem unloading via an apoplastic pathway enabling their transport from source to sink. The gene encoding an invertase Nin88 from tobacco was cloned and shown to be characterized by a specific spatial and temporal expression pattern. Tissue-specific antisense repression of Nin88 under control of the corresponding promoter results in a block during early stages of microsporogenesis, causing male sterility without any other morpho-physiological changes (Goetz et al., 2001). Whereas, exogenous supply of carbohydrates was found partially reversing this effect and opened up the possibility of maintaining this male sterility system. Fertility restoration can theoretically be achieved by crossing this GMS system with transgenic plants expressing distantly related



invertase (may be from bacteria), which is not under the control of the antisense repression of a plant invertase.

E. Transgenic Induction of Mitochondrial Rearrangements for CMSStability of the mitochondrial genome in plants is found to be controlled by nuclear loci by suppressing illegitimate mitochondrial DNA rearrangements during development (Sandhu et al., 2007). One such nuclear gene is Msh1, disruption of which leaded to the type of mitochondrial DNA rearrangements associated with naturally occurring cytoplasmic male sterility in tomato and tobacco. The male sterility showed maternal inheritance pattern in which segregants failed to reverse the male sterile composition rendering it stable, non-transgenic male sterility. Yang et al. (2010) also disclosed how a novel chimeric open reading frame gene ‘orf220’ when expressed via mitochondrial-targeting sequence of the β-subunit of F1-ATPase (atp2-1) downregulates the genes related to pollen development, thus causing male sterility in Mustard. Although reproducibility of this system is in question, it provides a method to develop novel protoplasm male sterile lines for unleash as non-GMO or non-transgenic materials.

F. Engineering CMS via Chloroplast GenomeRuiz and Daniel (2005), while investigating expression of the polyhydroxy butyrate pathway in transgenic chloroplasts, addressed the specific male sterility inducing role of β-ketothiolase encoded by phaA gene. They studied the effect of light regulation of the phaA gene under different photoperiods which shows successful expression in transgenics only via the chloroplast genome using the psbA promotor and 5’ untranslated region. The β- ketothiolase was efficiently expressed in all tissue types examined, including leaves, flowers, and anthers. The transgenic lines were normal but male sterile with collapsed morphology of pollen due to an accelerated pattern of anther development affecting their maturation. Reversion of the male sterility occurred under continuous illumination giving viable pollen and a good seed set.

G. Male Sterility through Site-Specific Recombination TechnologyAn expedient approach of genetically engineered

male sterility system is evident by use of site-specific recombination technology that generally contains a recombinase and its specific recognition sequence. Common site-specific recombination system includes Cre/loxP and FLP/FRT. The plants are transformed with the gene construct containing male sterility gene expression box flanked by two loxp sites in the same direction recognising Cre-recombinase and male sterile line is produced. Similarly, in another transformation experiment cre gene inserted expression vector of plants is used to yield restorer lines. In absence of cre gene, male sterility gene expression box within loxp sites produce male sterile phenotype which on hybridization with cre gene containing restorer line get reversed to male fertility due to cutting off the same male sterility expression box by the cre-recombinase encoded by introduced cre gene. This system was initially used by Bayer and Hess (2005) in tobacco for stilbene synthase (sts) transgene inducing pollen sterility while the successful attempt for Barnase gene was carried out by Cao et al. (2010) in eggplant.

These are the different approaches aroused or ought to be called forth in near future of inducing transgenic male sterility in crop plants. Acquiring as a transgenic or GMO (Genetically Modified Organisms) status by the final products of these systems raises the regulatory and legislative problems in a number of countries concerning their environmental safety issues. Hence aside these approaches, many other novel systems are also being evolved that employs ‘transgenic construct driven non transgenic male sterility’ systems as like SPT (Seed Production Technology), MCS (Multi-Control Sterility), etc. furnishing the final product as non-transgenic or non-GMO and being extensively applied at commercial level for hybrid seed production of some crops, e.g. maize. Even instead of the transgenic status issue, genetically engineered male sterility systems also encounter the other difficulties viz. non-availability of efficient gene construct, possible dispersion of transgene to other related species, prerequisite of efficient transformation and tissue culture techniques and very high initial cost investment. May the further advancement in the field of molecular biology and biotechnology erase these problems and ease the creation of cost-effective genetically engineered male sterility systems at the serve of hybrid seed production in crops.

20153

54. AnInnovativeTechnique:ArtificialSeedTAMILZHARASI, M.

Ph.D., Scholar, Centre for Plant Breeding and Genetics, TNAU, Coimbatore *Corresponding Author Email: [email protected]

Artificial seed or synthetic seed is defined as encasing of somatic embryos, shoot bud, or aggregates of a cell of any tissues artificially which has the potential to establish

a plant in the either in-vitro or in-vivo environment. Initially, synthetic seeds applied to somatic embryos with high economic value. Later this technique has



been introduced to other vegetative parts such as organogenic or embryogenic calli and shoots tips. This led a new path to link with other innovative and novel techniques like micropropagation which includes organogenesis and axillary bud proliferation. The outcome of the above techniques has resulted in naked structures, which are highly vulnerable to desiccation and biotic stress. Here is a need for technique like a synthetic seed that can overcome the above drawbacks by protective coatings and would also help to plant on a larger scale and increase the germination rate under the natural environment. Though the encapsulation of micro propagules is considered as a foremost technique that gives protection, it can also disseminate the propagules that have been developed in vitro.

Significance � Artificial seeds play a vital role in sustaining and

preserving the biodiversity of plants which can be benefited by other organisms through mutualistic effect.

� It is an essential technique to proliferate the plant species which are unable to set seeds (vegetative propagation) like seedless watermelon or grapes.

� Seed multiplication process in male or female sterile line as a parent in hybrid seed production technology. The conventional method of seed production in sterile lines incur more labour, resource, and time-consuming approach.

� Artificial seed technology provides a new avenue in the field of plant breeding and biotechnology. It helps in reproduction of transgenic plants by a somatic cell of a trait of interest and this could be useful to locate all the plants produced from a gene of interest.

� Reproduction of polyploids with elite traits was difficult in conventional breeding as the recombination takes place, but the is traits could be preserved using artificial seed technology.

� The advantage of long-term storage with vegetative reproduction of artificial seed has a huge impact on agriculture. The crops used in artificial seed production are categorized into two classes viz., i) Crops with high-quality somatic embryo and ii) Towards the application of commercialization.

� Synthetic seed resembles a zygotic embryo in terms of its components like seed coat and endosperm which resulted in easy handling and storage than the vegetative propagules.

� Apart from encapsulation, it is supplemented with growth hormones, plant growth-promoting microorganisms, and nutrition to increase growth and development of the somatic embryo to plants.

Advantages over other Methods of PropagationSynthetic seed or artificial seed has added advantage over micro and clonal propagation. They are large-scale and high volume, propagation method, maintain genetic uniformity of plants, low cost per plant, rapid multiplication of plants, and direct delivery of the propagules to the field (Bhatia and Bera,2015).

Classification of Synthetic SeedDepending upon the technology used to produce the synthetic seed, it is classified into two types,1. Desiccated synthetic seed2. Hydrated synthetic seed

Desiccated Synthetic SeedThere are two methods used in desiccation either slow or rapid method. In a slow method, desiccation is achieved in one or two weeks in a chamber with low relative humidity. Whereas in the rapid method, Petri dishes are left free from sealing overnight. It is applied to somatic embryos with desiccation-tolerant types. This method has been used and standardized in a wide range of temperate to tropical origins of a plant cell in several species by Engelmann et al., (2008).

Hydrated Synthetic SeedEncapsulation of somatic embryos with hydrogels (sodium alginate, potassium alginate, carrageenan, sodium pectate, or sodium alginate with gelatine). Recalcitrant types of somatic embryos are used. Nature of hydrogels should be nontoxic to the embryo, easily soluble in water and dried to form a thin film, does not support the growth of microorganisms (Kitto and Janick, 1985).

Part to be used for the Production of Synthetic Seed

Somatic embryosAmong the various parts of the plant, somatic embryos are widely used in synthetic seed production. Since they have advantages like the presence of both plumule and radicle to produce shoot and root, respectively. It maintains the uniformity of the genotypes or any other plant (clonal propagation) whereas in the sexual reproduction it results in variation among the progeny by the process of recombination. Site-directed changes in superior cultivar can also be obtained through somatic embryogenesis.

Shoot Buds and Shoot TipsPresence of unipolar nature of shoot buds and shoot tips with lack of root meristem. After regenerating the roots, the encapsulation process takes place.

Embryogenic massesTo produce the clonal plants and genetic transformation studies, embryogenic masses are used. It is a laborer intensive process to maintain the culture which requires periodic transfer to fresh media.

ProtocormsEncapsulation of propagules (protocorm) of orchids led to a higher multiplication rate than the normal vegetative propagation. It gave healthy plantlets when it was transferred to nutrient medium or natural environment portrayed by Corrie and Tandon, 1993 in Cymbidium giganteum.



Requirements for the Production of Synthetic Seed

Gelling agentsAmong the gelling agents (agar, alginate, carrageenan, carboxymethyl cellulose, guar gum, sodium pectate, gum tragacanth) alginate is used widely due to the advantage of quick gelatinization, moderate viscosity, the spin ability is low, biocompatibility, low cost and a greater number of capsule formations.

Artificial endospermThere is a difference between zygotic and non- zygotic (somatic) embryos. Unlike zygotic embryos, somatic embryos lack seed coat and endosperm. To combat the deficiencies, it is supplemented with growth regulators and nutrients in an encapsulation matrix which act as artificial endosperm. These artificial seeds can be stored for long term at 4˚C even up to six months without any compensation in viability.

AdjuvantThe important role of adjuvant is inhibiting the microbial attack and desiccation. Adjuvants can be nutrients, microorganisms, fungicides, pesticides, and insecticides. For example, supplementing activated charcoal increases the vigor and conversion of synthetic seeds. The reason behind this is charcoal breaks the alginate by which it increases the respiration. It retains the nutrients in a capsule and releases at a slow rate to the growing medium.

Steps Involved in Synthetic Seed ProductionSelection of explants

↓Induction of callus in explants

↓Somatic embryogenesis establishment in callus culture

↓Production of matured somatic embryos

↓Large scale production of embryos

↓Induction of desiccation and tolerance

↓Encapsulation of matured somatic embryos

↓Either invitro germination or planting in the field.

Applications of Synthetic Seeds1. This technology is useful in the propagation of

endangered species, genetically modified plants, sterile plants, and hybrids.

2. It is easy to maintain because of uniformity in genetic and phenotypic levels.

3. Synthetic seeds are easy to handle, transport and mechanized sowing is also possible. It reduces the production and maintenance cost per plant.

4. The exchange of synthetic seeds to other countries is very easy. Since it is free from pest and disease infestation. It doesn’t deviate from any plant quarantine obligations.

ReferencesCorrie, S. and Tandon, P. (1993) Propagation of

Cymbidium giganteum wall, through high frequency conversion of encapsulated protocorms under in vivo and in vitro conditions. Indian Journal of Experimental Biology, 31, 61-64.

Engelmann, F., Gonzalez-Arnao, M. T., Wu, W. J., & Escobar, R. E. (2008). Development of encapsulation-dehydration. In B. M. Reed (Ed.), Plant cryopreservation: A practical guide (pp. 59–76). Berlin: Springer Verlag.

Kitto SL, Janick J. (1985). Production of synthetic seeds by encapsulating asexual plant embryos of carrot. Journal of the American Society for Horticultural Science, 110, 277–82.

20154

55. Optical MappingSOHAM HAZRA* AND SHOUVIK GORAI

Department of Genetics and Plant breeding, B.C.K.V, Nadia, West Bengal- 741252 *Corresponding Author Email: [email protected]

Optical mapping is a kind of gene mapping technique which is used for constructing high resolution from optical maps. Optical maps are single, stained molecules of DNA. It equips orders and genome wide information and can detect large structural variations

in the DNA. This method of mapping was originally discovered by Dr. David C. Schwartz in the 1990s. since its development Optical mapping has evolved through the course of time. Initially DNA molecules were put on molten agarose which had restriction enzymes



pre-mixed with it. Next application of electrostatic interactions was taken into account and due to this the resolution has increased a lot such that fragments from 30kb to as small as 800bp can be sized. Nowadays, use of nest generation system called ‘Nanocoding’ is used, that traps elongated DNA molecules in nanoconfinements and boosts the throuput. Optical mapping has its direct implications in human and microbial diversity studies.

Optical mapping differs in several aspects in relation to conventional mapping techniques. The major upper hand of optical mapping over restriction mapping is that the DNA fragment can be preserved and no recinstruction is needed. Another vital advantage is that it omits the PCR process since maps are directly constructed from genomic DNA molecules. However, to minimise the false positive multiple optical maps are constructed from the same genome.

The uses of optical mapping are still not fully utilised. Initially it was used to map the whole genome of bacteria, parasites and other lower organisms. To validate the bacterial genome optical mapping has also been used. But until now only David C. Schwartz lab has utilised the technique to map the higher organisms like mouse, human rice and maize.

The optical sequence cycle undergoes the following steps:

1. DNA barcoding2. Template nicking3. Gap formation4. Flurochrome incorporation5. Imaging6. Photobleaching7. Repeat steps 4-6

Advantages of Optical Mapping1. Minimal DNA sample required2. Singla molecule analysis.3. Avoids the time consuming and costly

amplification step.4. While most of the other mapping techniques

requires large amount of small sequence reads, optical mapping can assay large DNA molecules.

5. Omits the PCR process.6. Multiple sequences can be obtained from the

same DNA molecule to reduce the false positive.

FIGURE 1: Optical mapping cycle sequence

Disadvantages of Optical mapping1. High level of precision is required since single

molecule is sequenced.2. Multiple incorporation is difficult since

the flurochrome label is not removed after incorporation resulting in bulky labels.

20167

56. Brown Planthopper: A Serious Threat to Rice ProductionISHWARYA LAKSHMI VG1 AND BASAVARAJ PS2

1Ph.D. Scholar, Dept of Genetics and Plant breeding, College of agriculture, Rajendranagar, Hyderabad. 2Scientist, Plant Breeding, ICAR-NIASM, Baramati.

Rice (Oryza sativa L.) is the major dietary intake for more than half of the world’s population. Yield of rice is halted by both biotic and abiotic stresses. Nearly 52% of the global rice production is annually lost because

of the damage caused by biotic stress factors, of which 21% is attributed to insect pests attack. Though rice is infested by more than 100 species of insects, 20 of them are considered as serious pests since they cause



significant damage to rice crop. Among the notable serious insect pests, Brown Planthopper (BPH), Nilaparvata lugens Stahl (Homoptera: Delphacidae), is considered as being one of the most destructive insect pests causing significant yield loss in most of the rice cultivars of Asia.

In the field, all the growth stages of the rice plant are completely vulnerable to BPH. Attack by the insect leads to yellowing of leaves, stunting of plants, reduction in growth, vigour, number of productive tillers and grain-filling. Heavy infestation of the insect causes complete drying and death of plants leading to a condition known as hopper-burn. In addition to causing several direct injuries to plants, it also causes indirect damage by acting as a vector for major viral diseases viz., rice tungro virus, grassy stunt and ragged stunt virus.

BPH Insect BiotypesBased on their ability to infect and feed on rice varieties, four different biotypes of BPH have been identified. Biotypes 1 and 2 are widely distributed in Southeast Asia, while biotype 3 is produced in a laboratory present in the Philippines. Biotype 4 occurs in the Indian subcontinent and is the most devastating.

BPH Resistant Genes and DonorsUntil now, 38 BPH resistance genes have been identified from different resistance sources. The majority of the BPH resistance genes are mapped on six of the 12 chromosomes namely, chromosomes 2, 3, 4, 6, 11, and 12. The BPH resistance loci are clustered on three chromosomes with Cluster A on the long arm of chromosome 12 having eight loci, clusters B and D located on the short and long arm of chromosome 4 possessing 13 and four QTL/gene loci, respectively. This includes the recently identified BPH34, BPH30, and BPH36 resistance genes and cluster C on the short arm of chromosome 6 with six gene loci. Out of total BPH resistance genes, 20 genes were fine mapped and only eight were cloned and characterized. These cloned genes were found to be encoding coiled-coil nucleotide-binding site and leucine-rich repeat protein, CC-NBS-LRR protein, plasma membrane-localized lectin receptor kinases, B3 DNA-binding domain protein and CC-NBS-NBS-LRR protein, respectively (Ji et al. 2016).

Systematic breeding programs have led to the identification of several donors related to BPH resistance in both landraces and wild species of rice. Currently, major genes related to resistance have been identified from diverse cultivated as well as wild species of rice using classical and molecular approaches of genetics. Fourteen resistance genes have been identified from seven wild rice species, O. glaberrima, O. officinalis, O. australiensis, O. rufipogon, O. eichingeri, O. latifolia and O. minuta, while the remaining genes have been identified from cultivated rice species (Wu et al. 2018).

Candidate GenesBPH resistance genes that have been cloned tend to

encode several kinds of proteins such as lectin kinase, SCR domain, B3 DNA binding protein, and exocyst-localized unknown protein. Three BPH resistance genes namely, BPH26, BPH18, and BPH9 have reported to encode a member of coiled-coil–nucleotide-binding site–leucine-rich repeat (CC-NBS-LRR). This NBS-LRR protein family is concerned with the activation of the salicylic acid (SA) dependent and jasmonic acid pathways that intervene with the inhibition of sucking in the phloem sieve element and causes a high degree of resistance against different biotypes.

Approaches for Enhancing BPH ResistanceBreeding for BPH resistance had always been a great challenge for breeders and biotechnologists as the resistance to the insect is not well durable due to adaptability and the evolution of biotypes infestation. There are several approaches for incorporating and enhancing BPH resistance in elite cultivars. These include:

Molecular Breeding for BPH ResistanceSince a long time, several BPH resistance genes have been identified and transferred into elite susceptible varieties at IRRI, and a series of improved cultivars with BPH resistance were developed and released. However, the improved cultivars carrying single resistance gene lost effectiveness due to the evolution of new BPH biotypes. Hence, in order to develop new varieties with more durable and stable BPH resistance, stable introgressions and use of more genes, preferably pyramided into multiple gene lines is required.

Marker assisted selection and backcross breeding for introgressing BPH resistanceBased on the position of BPH-resistance genes on the chromosomes previously reported, recently SNPs, SSRs and InDel markers linked to related genes are designed and are being utilized to track the target genes in the segregating populations. A minimum of 6–8 backcrosses are usually required to completely recover a recurrent parent genome using conventional breeding methods, but MABC enables the procedure to be shortened to 3 or 4 backcrosses. Thus, MABC is an efficient method for introgression of resistance from BPH donor to an elite cultivar using repeated backcrossing technique.

Pyramiding of BPH resistance genesUsing MAS based conventional breeding strategy; advancement has been made in pyramiding two or more major BPH resistance genes into susceptible cultivars. In order to develop new cultivars with durable BPH resistance, along with gene pyramiding exploition of genetic diversity is equally important for ecological reasons. Moreover, multilines (NILs, ILs, or PLs) carrying different assortments of genes can also help in managing BPH populations to lower levels.

Other approachesEven transgenic lines resistant to BPH were also developed. There was significant inhibition to brown



planthopper by decreasing the survival, overall fecundity, retarding development, and reducing the feeding of BPH on phloem tissue by insect bioassay and feeding tests. Resistance to BPH was achieved using homozygous transgenic rice lines expressing GNA, developed by genetic transformation and genetic analysis-based selection.

ConclusionIn order to accomplish complete resistance to BPH, introgression and pyramiding major genes may provide durable resistance and improve yield potential of

cultivars. Development of functional SNPs associated with BPH resistance could also act as a powerful tool for improving elite rice cultivars for BPH resistance.

ReferencesJi H, Kim SR, Kim YH et al (2016) Map-based cloning

and characterization of the BPH18 gene from wild rice conferring resistance to brown plant hopper (BPH). Insect Pest Sci Rep 6:34376.

Wu SF, Zeng B, Zheng C, Mu XC, Zhang Y, Hu J, Zhang S, Gao CF, Shen JL (2018). The evolution insecticide resistance in BPH (Nilaparvata lugens Stal) of China in the period 2012-2016. Sci Rep 8:4586

20181

57. LathyrusaCropwithitsNutritiousandToxicEffectAMRITA GIRI AND MINU MOHAN

Department of Genetics and Plant Breeding, Indira Gandhi Krishi Vishwavidalaya Raipur (C.G.) *Corresponding Author Email: [email protected]

Lathyrus is a legume crop belong to family Fabaceae distributed worldwide with 187 species and subspecies. These Species are found in the Old World and the New World. The centres of diversity for Old World species is mainly in Asia Minor and the Mediterranean region However, only one species – Lathyrus sativus is widely cultivated as a food crop, while other species are used to a lesser extent for both food and forage, however In India grass pea is mainly cultivated as fodder crop, while Some species are used as ornamental plants for decoration purpose, especially the sweet pea (L. odoratus). Lathyrus sativus, commonly known as grass pea and also known as blue sweet pea, chickling pea, chickling vetch, Indian pea, white pea and white vetch. It is cultivated in Asia and East Africa and mainly in India, Bangladesh and Ethiopia due to its higher adaptability under adverse climatic conditions unlike other pulse crops we can grow lathyrus in adverse diverse environmental condition due to its droughty nature with low input as it not required any special care and management. It can easily survive in rainfed condition as well as in waterlogged condition. Beside these entire characters grass pea is rich in protein content, essential amino acids and vitamins. If we overlook the nutritional qualities of lathyrus we find following values for L. sativus, energy 362.3 cal, protein 31.6%, fat 2.7%, nitrogen-free extract 51.8%, crude fibre 1.1% and ash 2.2%. Aletor et al. (1994) reported on selected lines of three Lathyrus species. They found that crude protein averaged 32.5% (of dry matter). As we know increasing population of India is a big threat now a days and there is scarcity of food which provide sufficient nutrient for the human body, in such condition we need to grow such food components crops which are high in nutrients like protein, vitamins and several other nutritional supplements needed for a human body. It can be attained by Grass pea, as it is the richest source of

protein after soybean with other essential nutrients. It can be used as chapatti and dhal. In addition to all this benefits lathyrus leaves approximate 36-49kg/ha nitrogen for the succeeding crop and in this way, it also provides benefit to soil by maintain soil fertility. As coin has two sides of their own effect, there is also a different side which is presence of ODAP content in Grass pea which is leading a motor neuron disease called lathyrism (paralysis of lower limbs). identified in L. sativus by Bell (1962) when he found ninhydrin-reacting compounds in many Lathyrus species. The precursor b (isoxazolin-5-on-2-yl)-alanine (BIA) is responsible for biosynthesis of b-ODAP in young seedlings. ODAP is present in all tissues of L. sativus plants, irrespective of age or variety, but maximum content was observed in the leaf at vegetative stage and in the embryo at the reproductive stage. It has been reported that continuous consumption of grass peas may lead to paralysis in human and animal. However, attempts have been done to provide growers with varieties with low ODAP content led to the release of the variety Pusa-24, Further research efforts led to the production of six more varieties (LSD1, LSD2, LSD3, LSD6, Pusa-305, and Selection 1276) with low ODAP content (up to 0.2%). Subsequent some other varieties named Ratan (Bio L 212), Mahateora and Prateek were released with Low ODAP content of 0.06, 0.08 and 0.08 respectively. But this is still not sufficient for safe consumption of lathyrus, beside all of this we need to draw our attention toward the direction of producing improved varieties with zero ODAP content so that we can use the benefit attributes of this crop for human and animals, however many biotechnological works are running in this aspect and our scientist are trying to silent the gene of ODAP content, If we get success it will be the new invention in agriculture field.



FIG: Ratan

20187

58. Distant Hybridization in Crop PlantsVERSHA

Ph.D. Scholars, Dept. of Genetics and Plant Breeding, CCS HAU, Hisar-125004 *Corresponding Author Email: [email protected]

Distant hybridization raises the crosses between distant species, in compare to close hybridization, which occurs between varieties of the same species or subspecies. Distant hybridization can combine the biological characteristics of distant species, which breaks species limits and increases genetic variations, resulting in genotypic and phenotypic changes and the creation of new variations or species. The concept of distant hybridization has also been expanded to geographically distant hybridization. Thomas Fairchild in 1717 recorded the first authentic distant hybridization for the crop improvement in the production of a hybrid between Carnation (Dianthus caryophyllus) and Sweet willian (Dianthus barbatus). Many excellent crop varieties have been obtained and popularized in large areas, for instance, the distant hybridization of couch grass (Elytrigia repens), common wheat (Triticum aestivum), and rye (Secale cereale) resulted in the creation of new crop species that do not exist naturally. Distant hybrids contain the genomes of different species, laying the foundation for recombination and communication between the genes of different species.

Requirement of Distant Hybridization � Diseases and insect resistance and quality � Wider adaptation � Mode of reproduction

There are many difficulties or major interspecific crossability barriers encountered in distant hybridization like:

� Temporal and spatial isolation of species � Pre-fertilization barriers:

– Pollen –Stigma interaction– Pollen Tube-Style interaction– Pollen Tube- Ovule interaction

� Post fertilization barriers

– Non-viability of hybrid embryos– Failure of hybrid to flower– Hybrid sterility– Lack of recombinant

Techniques to remove the difficulties or crossability barriers in distant hybridization

� Somatic hybridization � Alien Addition lines � Alien substitution lines � Embryo Rescue � Somatic hybridization: Production of hybrid

plants through the fusion of protoplasts of two different plant species/varieties is called somatic hybridization and such hybrids are known as somatic hybrids. Klercker (1892) was first isolated plant protoplasts by cutting plasmolysed cells with a fine knife.

� Alien addition lines: These lines carry one chromosome pair from a different species in addition to the normal somatic chromosome complement of the parent species. The main purpose of alien addition is the transfer of disease resistance from related wild species to the required one e.g., transfers of mosaic resistance from Nicotiana glutinosa to N. tabacum.

� Alien substitution lines: This line has one chromosome pair from a different species in place of the chromosome pair of the recipient species. Alien substitution lines have been developed in wheat, cotton, tobacco and oats. The alien substitution shows more undesirable effects than alien additions and more useful in agriculture.

� Embryo rescue: When embryo fails to develop due to endosperm degeneration, embryo culture



is used to recover hybrid plants that’s known as hybrid rescue e.g., H. vulgare x Secale cereale. Embryo rescue generally used to overcome the endosperm degeneration.

Limitations of Distant Hybridization � Distant crosses are associated with problems of

cross incompatibility, hybrid sterlity and hybrid breakdown. It causes difficulties in interspecific or intergeneric gene transfer.

� Sometimes these hybrids have several undesirable characters such as non-flowering, late maturity and useless combination like Raphanobrassica

� Desirable characters are generally linked with some undesirable characters which cause difficulties in the use of desirable genes from wild species.

� Specials techniques are required to make distant hybrids.

Applications of Distant Hybridization � Development of new varieties (e.g.; Gossypium

hirsutum x G. barbadense hybrid varieties of cotton)

� Production of new crop species (e.g.; Triticale hexaploide)

20216

59. Role of Cell Signaling in Crop ImprovementMEENAKSHI RATHI

Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar 125004

World’s population is about 7.8 billion which is increasing at the rate of 1.2 % annually and to feed this population only 50 million sq.km land is being used to produce food thus we need to increase productivity to feed mankind. Since the green revolution conventional breeding methods played vital role in improving crop productivity but in present scenario it has to be supported by more advance technologies. In this article, role of cell signaling and how modification in the signaling pathways may be helpful for upcoming challenges in agriculture have been discussed.

Cell signaling is a biological mechanism that occurs in cell which gives cells an ability to receive or generate signals in response to their surrounding environment. Cell to cell communication between cells is mediated by extra-cellular signal molecules. Some of those operate over long distances, other signal only to immediate neihbrouring cells. Cell signaling is about communication between different group of cells and tissue…how one group of cells informs another group of cells what to do. Properties of cell signaling are specificity, affinity, cooperative, sensitization, integration and amplification. That means signaling molecules are very specific to trigger stimuli, action is well coordinate by all cells and integrated by different stimuli also signals are amplified for easy detection.

Cell signaling has various components viz. signaling molecule, receptor, transducer, secondary messenger, transcription factor. Signal molecules are the one which generate signal or act as stimuli. Molecules like hormones, elicitors, change in ion concentration act as signal molecules. These signal molecules are detected by receptors present on the cell surface of plant and transduce signal further. Receptors can be ion gated channel, R gene products, G-protein etc. R gene recognizes the elicitor of the pathogen and thus resistance response is triggered. When products are able to recognize the avr gene product, then they

induce resistance to that pathogen. The heterotrimeric G-protein complex is composed of Gα, Gβ, Gγ subunits and activated by cell-surface receptors called G-protein-coupled receptors. GTP binding initiates structural rearrangements which disengage the Gβγ and results in dissociation of heterotrimer. The free subunits then send signals through the interaction with downstream proteins, called effectors. Signal transducer amplifies the signal and activates transcription factors. Calcium-dependent protein kinases (CDPKs) are encoded by large gene families. CDPKs are regulated by the binding of Ca2+ to the regulatory domain (calcium activation domain or CAD). The Arabidopsis genome encodes a minimum of 34 putative CDPKs (Harmon et al., 2001). In rice plants, a membrane-associated CDPK was activated by cold treatment (Martin and Busconi, 2001) these finding shows the presence of CDPKs in crop plants. Pathogen/microbe-associated molecular pattern (PAMPs/MAMPs) and damage-associated molecular patterns (DAMPs) are detected by pattern recognition receptors (PRRs) on the external face of plant cells. PRRs then send signal to MAPK (Plant mitogen-activated protein kinase). MAPK induces multiple defense responses like stomatal closure, reactive oxygen species (ROS) generation, defense gene activation, hypersensitive response (HR) and cell death. VIP1, a bZIP transcription factor, is phosphorylated by MPK3, which consequently leads to the re-localization of VIP1 from the cytoplasm to the nucleus to activate the PR1 pathogenesis-related gene. There are 20 MAPKs, 10 MAPKKs, and approximately 60 MAPKKKs in the Arabidopsis genome based on sequence homology. (Hamel et al. 2006, Ichimura et al. 2002) and thus activation or deactivation of a transcriptional factor can be done in desired direction. MAPK and CDPK then activates different transcription factors like AP1, EREB, bZip which in turns up or down regulate PR genes, SAR inducing genes, DREB



gene (in dought stress) which triggers responses such as hypersensitive reaction, LEA protein synthesis, stomatal closure etc thus provides defense mechanism in plants.

Clear picture of cell signaling helps in enhancing defence mechanism of plants for example vivek et al. (2013) isolated ZoCDPK1 from a stress cDNA generated from ginger using RT-PCR technique and over-expression CDPK1 gene in transgenic tobacco conferred tolerance to salinity and drought stress accompanied with high percentage of seed germination, higher relative water content, expression of stress responsive genes and increased photosynthetic efficiency. Whereas in another study by Wang et al. (2017) gene silencing of GhMPK20 in cotton enhanced

resistance against fusarium wilt. These studies show we have ample of opportunities to improve crop resistance and productivity if we understand cell signaling pathways. Modification of the pathways at any stage according to the need will help in better response of plants against biotic-abiotic factors. The major advantage of the technique is it does not involve Trans genes hence no ethical and moral issues could be raised. Future cell signaling studies would involve integration of genomics, transcriptomics, proteomics and metabolomics data that will provide complete picture of cellular mechanisms and their responses. Thus, we will be able to manipulate the pathways to increase resistance against biotic-abiotic factors.

FIG: Systematics pathway of cell signaling during stress;



SEED SCIENCE AND TECHNOLOGY

20088

60. Homa Farming and its Uses in AgricultureISLAVATH SURESH NAIK

University of Agricultural Sciences, Dharwad

IntroductionAgnihotra refer to the daily twice doing homa and offerings made by those in the srauta tradition. This tradition dates back to the Vedic age; the bhramhans perform the agnihotra ritual chanting the versus from the Rigveda. The tradition now practiced in many parts of south Asia in the Indian sub-continent including primarily India and most particularly in Nepal. Origin is in all four Vedas like rig Veda, yagura veda, Sama veda and atharvana veda. Actually, it was not bothered and forget and in 1985 shri swami param pujya sadguru gajanam maharaj again started to tell about this spiritual practice. Homa farming has its origin from Vedas. Principle involved in it is ‘’you heal the atmosphere and healed atmosphere will heal you’ ‘It is called revealed science. It is an entirely spiritual practice that dates from Vedic period. The basic aspect of homa farming is the chanting of Sanskrit mantras (Agnihotra puja) at specific times in the day before a holy fire. The timing is extremely important

Types of Homa1. Agni hotra homa: It is most important and

practised exactly at sunrise and sunset timings only.

2. Vyahrithi homa: It can be performed at any time except sunrise and sunset. It is also performed at when commencing at om trayambakam homa.

3. Om trayambakam homa: It should be performed at least 4 hours a day. It should be performed 24 hours on full moon and no moon day.

AgnihotraAgnihotra is the basic Homa fire technique, based on the bio rhythm of sunrise and sunset, and can be found in the ancient sciences of the Vedas. Agnihotra has been simplified and adapted modern times, so anybody can perform it. During Agnihotra, dried cow dung, ghee (clarified butter) and brown rice are burned in an inverted, pyramid-shaped copper vessel, along with which a special mantra (word-tone combination) is sung. When the mantras are chanted in the meter in which they are composed, by Supreme Grace the inherent meaning and the power and vibrations impact the entire creation. The healing occurs at the grass roots level in the subtlest manner. This power of the mantra is locked into the ashes that develop in the fire upon the oblations. These vibrations pulsate the entire universe in a profound subtle but sure impact and

affect. It is like fragrance and beauty of a flower when the flowers are in the form of a bud. On uttering the word Swaaha and the offering in the fire a phenomenal nourishing and energizing impact is created on the entire life and creation.

How to perform agni hotra1. Smear few cow dung chips with ghee and arrange

them in the Agnihotra pot.2. Mix about a teaspoon full of brown rice with a

small amount of ghee and keep them aside.3. Start the fire few minutes before sunrise/sunset.a) While chanting the mantra offer the rice smeared

with ghee (just enough that one can hold in the tip of five fingers) at the utterance of ‘Swaaha’in the fire.

b) There are only two offerings at Sunset or Sunrise each in the Agnihotra fire.

Agni hotra mantraAgnihotra mantra MeaningAgnaye swaáhá, Unto the fireAgnaye idam na mama I am offering all.Prajápataye swaáhá, This offering is not minePrajápataye idam na mama it is Thine.

Effect of agnihotra on atmosphere � Homa applications are a practical contribution to

environmental protection because they purify the atmosphere and improve the quality of air, water and soil.

� The effects of so called subtle (vital) forces and energies on plants, as well as an humans and animals.

Effect of agnihotra on soil � Increases the availability of soluble nitrogen. � Stabilizes the amounts of potash, nitrogen and

trace elements in it. � Increases the amount of water soluble

phosphorous available to plants in the soil, this has great effect on their growth and reproductive cycles.

� Earth worms proliferate in the environment, due to their increase in their hormonal production they distribute moisture in the soil and provide it with humus.



Impregnation of seeds and bulbsMixture of cow urine and water in a ratio of 50:50 to which up to 4 table spoons of agnihotra ash per 5 litres of solution are added and stirred. Seeds and bulbs should soak in this solution for 30-40 minutes. This strengthens the germinating plant and makes it more resistant to pests. Like cow dung cow urine has antibacterial effects and provides a protective coating around the seeds and bulbs. After this time of treatment seeds are spread on filter paper, or other absorbent paper, to dry. They should be dry enough to spread, but moist enough so that the core of the seed doesn’t dry out. Bulbs may be planted immediately after treating with the solution.

FertilizersIn addition, plants can be fertilized with a mixture of agnihotra ash, stinging nettles and water. The special liquid fertilizer strengthens plants. The stinging nettles are fermented i.e. decomposed in the water for 7-14 days, depending on the weather conditions and the number of nettles needed. This mixture should then be diluted to a solution with a ratio of 1:9 in other words,1part stinging nettles solution is mixed with 9 parts of water and filtered with a fine screen [sieve] in to a spraying container or watering can.

Plant nutrient solutionTo make an Agni hotra plant nutrient solution, up to 4 table spoons of agnihotra ash and up to 4 table spoons of pulverized dried cow dung are stirred in approximately 5 litres of water and then applied to plants. This may be repeated every 14 days, depending on how much it needed.

Spray solutionA nutrient solution to be sprayed can be made by mixing up to 4 table spoons of Agnihotra ash with 5 litres of water. This spray solution is left standing for 3 days and then filtered through a fine screen before it is used to protect plants against pests and diseases. A spray solution can also be made from certain fern blossoms, in which approximately 150 grams of blossoms, mixed with 2 litres of water and 2 table spoons of Agnihotra ash, are left standing to ferment for 7-10 days. Filtered through a narrow-meshed screen and then finally distributed on the plants with a sprayer. This helps keep away pests such as snails.

Gloria biosol an effective homa bio fertilizerGloria biosol is a very effective bio fertilizer which can be produced simply in homa atmosphere. Biosol liquid can be used for foliar application to nourish plants and soil. Biosol is superior to vermiwash as it contains high number of beneficial microorganisms and energy of homa process. Agnihotra ash has a significant positive effect on all the materials used and makes the biosol rich in macronutrients.

Materials are mixed in a large tank (200, 500 or 1000 litre). One copper Shree Yantra disc is placed in the tank. The tank is then sealed and kept for 20

to 30 days. After digestion is complete, the slurry can be removed. Biosol is used diluted with Agnihotra ash water solution in the ratio of 1:10. For one hectare of agricultural area, 200 litres of Biosol in solution are required. Biosol in solution can be sprayed on any type of crop at an interval of fifteen days. The application of the Biosol solution should be made before sunrise or after sunset. We can preserve Biosol liquid in air tight cans it will last longer, say about six months. Left over solid Biosol which is having maximum macronutrients should be mixed with any type of organic manure at a ratio of 1:5.

Agricultural benefits of agnihotra ash � Faster germination rate with the application of

agnihotra ash. � Higher yield of crops. � Green and healthy crops. � Resistance to pests and diseases. � Early blooming of flowers. � Crops are fertilizer or chemical free.

Conclusion � By using homa practice the environment in which

it is done is free of diseases and pest attack to crops grown there.

� It energises the area it is practiced. It also heals the environment where it is practiced.

� Homa ash obtained also supplies nutrients to the plants and thereby giving high yield and high-quality produce.

ReferencesBrunda, R., Basarkar, P.W., Dharmatti, P.R., Patil,

A.A., and Babalad, H.B., (2011) Response of tomato (Solanum lycopersicum L.) to Homa Organic Farming Practices. Journal of Applied and Natural science.6(4);213-218.

Hattalli Sachin Amrutraya, Basarkar, P.W., Dharmathi, Math and Sreenivasa., (2011) Biochemical studies on Homa organic farming practices in cabbage (Brassica oleracea var. saint) Journal of Applied and Natural science.6(6);210-214.

Kumari Namrata, Bhaskar, P.W., Babalad, H.B., and Sreenivasa, M.N., (2012) Yield and yield attributes and economics of soyabean as influenced by homa organic farming practices. Karnataka J. Agric. Sci.25 (2):270-272.

Kumar R, Kumar A., Chakraborty, S. and Basarkar P.W., (2017) Effect of Homa Organic farming on growth yield and quality parameters of Okra. Journal of Applied and Natural science. 9(4):2205-2210.

Rajeev Kumar, Pushpa Singh, R.K., Rai, Mala Kumari, Adyant Kumar, A.D., Pathak and Nagaratna S Olekar., (2018) Homa organic farming reduces pest and diseases infestations in okra and improves its quality and yield. Journal of Entomology and Zoology studies. 6(6):211-218.



20122

61. Diagnosis of Seed Borne PathogensSUNIL KUMAR

Ph.D. Scholar, Department of Plant Pathology, SKN College of Agriculture, SKNAU, Jobner, Jaipur (Rajasthan) - 303329 *Corresponding Author Email: [email protected]

IntroductionSeed borne pathogens we now that it has greater impact on agriculture as well as seed industries and detection of seed borne pathogens is very much essential as the seed borne inoculums may give rise to severe disease investigation in the field condition and severe loss to the growers. There are conventional methods as well as molecular methods that are deployed for the detection of diagnosis of seed borne pathogens. Some of them are like:

Methods1. Blotter method2. Agar method3. Paper towel method4. Embryo Extraction method5. De-hulling and embryo extraction6. Extraction and Agar plating method7. Extraction and Polymerase Chain Reaction

method8. ELISA method

These are some of the common seed borne methods applied for seed health testing for detection and diagnosis of seed borne pathogens.

Blotter method: Say for example Alternaria dauci that causes disease in Daucus carota (carrot) and it’s a seed borne pathogen. It can be diagnosed with Blotter method. In this particular method we take 3 layers of blotting paper on both the lids of petriplate and the we moisten it and then we place the seeds at uniform distances and then we incubate it for period and then once pathogens grow outside the seed we can visualize the pathogens under microscope at different magnifications. And then by looking at the morphology of the fungal pathogen associated with it we can very well identify the pathogenic nature of and the pathogens associated with the carrot seeds. So, in short three layers of 90mm filters are placed in both the lids after soaking with sterilized distilled water. We have to drain away the excess water.

Then we should place at least 10 seeds on the plate, evenly placed, on the surface of the filter pare, then incubate it for 3days at 20oC in the dark. Then transfer the plate to freezer and maintain a temperature of -20oC for 24 hours and after freezing incubate for 6 days at 20oC with alternating 12hour period of darkness and near NUV lights and plates should be approximately 25cm below the lights and should not be stacked. Then we can examine as we have shown in the previous slide, the fungal spores or fungal growth on the seed

surface and we can establish the relationship between the inherent thing pathogen like fungi that is on the carrot seeds following this particular method.

The same Blotter method is also used for detecting seed borne pathogens such as Alternaria radicina in carrot, Botrytis cinerea in sunflower seeds. Then Alternaria radicina in carrot can also be established through malt agar method.

Malt agar method can also be diagnosed through Leptosphaeria maculans and Plebdomus biglobosus in Brassica seeds, then Ascochyta pisi in Pisum species and the method is basically Aseptically place around 10 seeds evenly spaced on the agar surface of malt agar plate. Then incubate it for 10days under 20oC with altering 12hour periods of darkening and near UV light (NUV). The similar way plate should be approximately 25cm below the lights and plate should not be stacked. Subculture of reference culture to a malt agar plate at the same time of the seeds are plated and incubated in the test plates so that we can have a comparative idea between the actual culture and the fungal growth that is taking place on the seed surface. Then examine the plates visually under stereoscopic microscope to establish the morphological characteristics of the associated fungi.

Colletotrichum lindemuthianum in bean then Bipolaris oryzae in rice these are some of the other pathogens that are also used in agar plate method for detection as seed borne pathogens.

Then the next method is Rolled paper towel method where seeds are placed between two paper towels and paper towels were moistened and the seeds were allowed to germinate and grow as seedlings and by looking at the seedling health one can establish from the germinating seeds of infected and healthy seeds then looking at the fungal spores or fungal colony strata associated with the infected seeds to establish the relationship between fungi that is causing certain seed borne diseases.

Embryo Extraction method - it is used for Ustilago nuda in case of barley. So the method basically as such we have to place the seeds 1 litre of freshly prepared 5% aqueous solution of sodium hydroxide and maintain at 20oC for 24hours. After soaking, the entire sample should be transferred to a suitable container and washed in warm water to separate the embryos, which appear through the softened pericarps. Then collect the embryos in a sieve of 1 mm mesh. Additional sieve of larger mesh can be used to collect pieces of endosperm and chaff. Then transfer the embryos to a mixture of equal quantities



of glycerol and water in which further separation of embryos and chaff can be made and transfer the embryos to a beaker containing 50ml of lactic acid solution and clear them by maintaining the lactic acid solution at boiling point for approximately 5min in a fume cupboard. Then transfer the embryos to fresh glycerol for examination. The scutellum becomes more transparent when embryos are left in glycerol for 1-2 hours making the examination much easier.

So, this is how we can go for Embryo Extraction method for detection of Ustilago nuda in barley seeds. Then we have to examine it under microscope for presence of the Ustilago nuda pathogen. Ustilago nuda in barely seeds can also be done by dehulling and embryo extraction method. Here, the embryo extraction method is the same but the dehulling method has to be preceded the embryo extraction method. In case for dehulling place the working sample in glass beaker with 25-37% sulphuric acid until the seeds are covered. Incubate in an oven at 75oC for 50min or until the seeds turn a medium brown colour. Carefully pour of the sulphuric acid solution, rinse seeds by pouring water into the beaker, gently mix and pour off the water and add new water and remove loosened hulls by stirring robustly with a rod. Remove hulls by carefully removing the water. If hulls remain, add new water and either use a hand mixer at low speed or continue stirring. Repeat the procedure until all hulls are removed. Be careful not to lose any kernels i.e. seeds without hulls. So, after dehulling we can go for embryo extraction method as it is mentioned earlier.

The next stage Extraction and Polymerase Chain Reaction method - So, this is used for detection of Xantomonas campestris, pathogonas campestris in Brassica seeds. Here the pathogen is first cultured

on petriplates and then DNA is extracted from the bacterial cultures and PCR amplification is done using specific primers for detecting the pathogens. Presence of specific bans on the cultures confirms whether the pathogen is of the targeted ones or and other microbes that is associated with. The detail about Polymerase Chain Reaction method will be discussed in subsequent talks. Similarly, Elisa method is used for detection of certain viral diseases of that are seed borne in nature for example squash mosaic virus, cucumber green mottle mosaic virus and melon necrotic spot virus in cucurbit seed.

So, another test that is known as Grow-out test. So this infected seeds are grown on sample pots and they are allowed to grow in a way that disease symptoms are manifested on the leaves. So, this is another confirmatory test which we call it as Grow-out test. It can be also perform for Squash mosaic virus where the symptoms are very much evident after growth of the seedling to a certain stage and by doing this test by looking at the symptoms one can confirm whether the pathogen is associated with the seeds or not. So, these are some of the basic methods for detection and diagnosis of seed borne pathogens and with the time different molecular tools have been deployed for detection of seed borne pathogens more accurately along with the races and strains that are associated with the disease seed lot and finally we can conclude that with this methods we can certainly able to establish a seed lot whether it is affected or not with certain fungal bacterial or viral pathogens.

Suggested ReferencesAgarwal, V.K. and Sinclair, J.B. 2014 (IInd Edition).

Principles of Seed Pathology. Levis Publishers.Karuna, V. 2011. Seed Health Testing-Principles and

Protocols. Kalyani Publishers.

20145

62. X-Rays in Seed Science and TechnologyC. TAMILARASAN* AND L. ANILKUMAR

Department of Seed Science and Technology, TNAU, Coimbatore, Tamil Nadu - 641003. *Corresponding Author Email: [email protected]

IntroductionX-rays is a form of electromagnetic radiation which is similar to light but with shorter wavelength and capable of penetrating solids and ionizing gases. The wavelength ranges from 0.1 to 10 nm. It was first discovered by Roentgen in 1895 who described that radiation is invisible but it can produce visible images through fluorescent screen. It is one of the faster and non-destructive method which is used in seed analysis. X-rays with higher photon energy and shorter wavelength are called hard X-rays whereas X-rays with lower photon energy and longer wavelength are called soft X-rays.

PrincipleX-rays are generated from X-ray tube, when a beam of electrons are striking the target at higher voltage. X-rays produced have different wavelength and different penetrating powers according to its accelerating voltage.

Production of X-RaysX-rays are produced from the X-ray tubes and the most used X-ray tube is modified Coolidge tube. It consists of cathode which is negatively charged and a positively charged particle termed as anode. The cathode is made up of tungsten filament which emit electrons when



heated. The anode consists of a target material which is made up of tungsten or molybdenum embedded in the copper bar. Water circulation is regulated around the anode in order to cool it when it’s get heated. The target is placed at an angle relative to the outcoming beam of electron. The electrons emitted by the cathode are accelerated by the anode and acquires higher energy. When the target stops the electrons suddenly, X-rays are emitted. The magnetic field associated with the electrons undergoes a change and X-rays are produced in the form of electromagnetic waves.

Properties � They travel in straight lines. � It cannot be deflected by electric field or magnetic

field. � They have high penetrating power. � It causes glowing in exposed fluorescent material. � They have the ability to produce photoelectric

emission and ionization of gas.

Invisibility to VisibilityAs the X-rays were invisible electromagnetic radiation it creates the visible image which is needed for our evaluation. The production of X-rays is related to the number of accelerated electrons. This determines the contrast of image produced when viewed under X-rays. In older days, barium platinocyanide screen was used to absorb the X-rays and which convert radiation to photons through a process called fluorescence. Negative flims and photographic paper were used in later days to make the X-rays into visible images and it also has the ability to store the images. At 1990’s a new material called scintillators was used to convert ionizing radiation into light. Through these scintillators high quality images can be visual at lower exposure of radiation. Nowadays these scintillators are coupled with digital camera which resulted in creating signals to provide a digital image that can be analysed and presented through computer.

Image FormationIn 2-dimensional projection of X-rays image is formed based on the amount of X-rays that is transmitted through seed. In 3-dimensional projection, seed is

placed between the X-ray source and the detector. The image is displayed based on the orientation of the seed and density distribution inside the seed. In 3-D X-ray imaging, the first step involved is interaction of the X-ray with object of seed structure and detection. The second step deals with reconstruction of 3-D structures from the data and resulted in tomographic images of object.

X-Rays - A Non-Destructive AnalysisThe first seed analysis done using X-rays is carried out in coniferous tree seeds by Lundstorm in 1903. X-rays are the potential tool to examine the seed not only in the Quiescent state but also in the activated state. This is a non-destructive and rapid method for analysis.

� X-ray radiographs gives the detailed information about the degree of insect infestation during storage or impact of damaging during field drying, threshing, harvesting and processing.

� In seed processing, effectiveness of a machine only decides the quality of the seed. Therefore, X-ray imaging of the seed lots can be used to adjust the machine parameters inorder to get the good quality seed.

� Seed characteristics such as embryo, endosperm development, size and position of cotyledons, seed coat thickness can be visualize through X-ray radiography.

� X-ray scanning is helpful in identifying the empty seed lot in pasture seeds.

� X-rays are used to inspect the embryo thickness in primed seed and to know thickness and homogeneity in seed coating and pelleting.

� X-rays are used to detect the polyembryony seeds in several crops.

� The level of hybridity in produced seed lots can be identified through X-ray scanning which is useful in rejecting selfed seeds present in the lot.

� Seed physical properties like shape, size and colour can be visualized by the X-rays which is useful in seed multiplication, maintenance breeding or in new varietal identification.

� The physical properties of the seed in different varieties analysed using X-ray radiography is also used in varietal identification.

20147

63. Genetic Use Restriction TechnologySRIDEVI RAMAMURTHY

Department of Agriculture, School of Agriculture and Biosciences, Karunya Institute of Technology and Sciences, Karunya Nagar, Coimbatore 641 114, Tamil Nadu, India. *Corresponding Author Email: [email protected]

IntroductionGenetic use restriction technology (GURT) also known as terminator technology is the genetic modification

and activation of genes to produce the sterile seeds, thus also called as suicide seeds. In the 1990s, the terminator technology was developed by United States Department of Agriculture and Delta and Pine Land



company. In 2002, Monsanto acquired Delta and Pine Land company and also termed this technology as gene protection technology. This technology switches the embryo of the second-generation seeds to be sterile and prevents the farmers to reuse the seeds that tends them to purchase new seeds for every sowing season.

Genetic Use Restriction TechnologyThe GURT process uses the components such as target gene, promotor gene, blocker sequence, trait switch and gene switch. The plants that possess GURT contains target gene triggered by the promotor gene. The promotor gene is prevented from target gene by blocker sequence. The gene switch after receiving the external stimuli amplifies the input and translate it into biological signal. The biological signal is received by the trait switch and an enzyme is created to cut the blocker sequence. The promoter gene can make target gene to express after elimination of blocker sequence. In other process, an operator has to bind with trait switch that makes the enzymes to cut the blocker sequence. This process is also prevented by binding of repressors with trait switch. Once the external input is introduced, in the place of trait switch, repressors bond with it that allows the enzymes to cut the blocker sequence and thus the trait is expressed.

Types of GURT � V-GURT � T-GURT

V-GURTVariety specific genetic use restriction technology (V-GURT) is related to plant variety level and results in production of sterile seeds. The target gene gets activated when the plant attains reproductive stage. The target gene also known as disrupter gene is a cytotoxin that denatures DNA in the plant and affects the normal functioning of seeds and seeds do not germinate. Chemical inducer triggers the genetic process that inhibits the seed development process if the seeds are replanted. The second-generation seeds will be sterile and thus farmers are unable to store it till next sowing.

T-GURTTrait specific genetic use restriction technologies that are restricted to trait level and thus known as T-GURT. This technology is considered as second generation of V GURT. T GURT affects the traits like germination, growth, flowering, tolerance level with the help of promoters. The transgene expression is regulated by promoters by the use of recombinase or gene silencing method. The genetic changes made in the crop can be

altered by chemical treatment and other factors like heat. Unless farmers buy the chemical activator, they could not acquire the enhanced trait incorporated in the crop.

Advantages of GURT � Reduces propagation of volunteer plants due to

production of sterile seeds. � This technology can be used to limit the spread of

genetic modified crops. � Prevents V GURT sprouting that reduces the seed

quality. � Prevents escape of transgenes into wild relatives

and reduces impact on biodiversity. � Protects the intellectual property rights of biotech

companies. � It is advantageous to the private firms, as the

farmers could not able to reuse the seeds.

Disadvantages of GURT � Due to production of sterile seeds, which makes

the seeds useless. � As the chemicals treated before sowing, the seeds

may be hazardous to animals and humans. � The plants may transfer pollen grains to other

wild crops and make them sterile too. � The toxic gene can be activated by chemicals

which affects the soil microorganisms. � Inadequate inducing agent to activate the

terminator genes may not trigger the genes and results in non-germination of seeds in next generation.

ConclusionTerminator technology may offer considerable incentives in agricultural sector for increased private sector innovation. It will also have adverse significance on conservation and development of plant genetic resources. Genetic use restriction technology has both good and negative impacts on agricultural systems. The developed countries do not have much impact on farmers but in developing and under developed countries terminator technology may affect farmers.

Referenceshttps://www.sites.ext.vt.edu/newsletter-archive/

cses/1999-02/1999-02-03.htmlhttps://en.wikipedia.org/wiki/Genetic_use_

restriction_technologyhttps://www.slideshare.net/siddarudh/gurt-geneetic-

usehttps://www.biotecharticles.com/Agriculture-Article/

Terminator-Gene-Technology-Types-Advantages-and-Disadvantages-4123.html



20183

64. Halogen Dry Seed TreatmentN. VINOTHINI1, POOVARASAN T.2,

BHAVYASREE R. K.3, AND M. SAKILA4

1Teaching Assistant, Agricultural College and Research Institute, TNAU, Eachangkottoi, Tamil Nadu – 614 902 2Ph.D. Research Scholar, Dept. of Seed Sci. Tech, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu – 641 003 3Assistant Professor, Agricultural Research Station, Kerala Agricultural University Mannuthy, Kerala – 680 651 4Assistant Professor, Agricultural College and Research Institute, TNAU, Eachangkottoi, Tamil Nadu – 614 902

Seed is a biological element and it’s ageing in an evitable process earlier physiological development whether the seed is in mother plant or it storage. During seed storage rapid loss of vigour and viability of seed is one of the significant constraints looked by the seed business and relating financial implications. Seeds stored under ambient condition, ageing of seeds cannot be arrested completely by adoption of suitable storage technologies it can however be controlled to an applicable extent. Seeds are dry dressing with halogen has conferred impact by bringing down lipid per oxidation and there by extension of seed vigour and viability during storage. Seeds when treated with halogen reduce physiological and pathological deterioration and slow down the deterioration senescence in a number of crop seeds.

SeedImportant in seed performance implies to a seed invigoration by any post-harvest treatment that concentrated on enhancement in seed germination, seed storage and better performance. A qualitative improvement in the seed should bring by seed invigoration, which should persevere much after the treatment and treatment impact essentially be physiological in nature.

Definition of HalogenationsHalogenation is a chemical method that happens when hydrogen atoms are supplanted by a halogen in an organic compound such methodology is called as halogenation. The elements fluorine, bromine and chlorine are present in halogens. Halogenation is to balance and alleviate the production of free radical that accelerated the deterioration of seeds.

Purpose of HalogenationsIt gives a security component to the seed ideally directly in the start of seed storage, which will lighten deteriorative seed senescence during storage. This treatment could likewise broaden their protection against the outward factors of seed deterioration specifically seed pathogen and insects.

Significance of Seed HalogenationsThis treatment stays away from utilization of water. Extended storage ability does not require drying back to safe moisture content. Mainly used for large scale application. This treatment likewise has the extra cost- saving advantage that will occur if the dry treatment is given before the seed is bagged and stored. In halogenations treatment, seeds are directly exposed to halogens like chlorine, bromine and iodine or through the carriers.

Methodology of Incorporation Of HalogensThe carrier is first exposed to the halogen in vapour in optimum quantities. The vapour saturated carriers are dry dressed with the seeds. These chemical compounds are likewise added directly to the carrier.

Anti-Microbial Property of HalogensAnti-microbial properties of halogen serve as a principle factor in extension seed storability. Unsuitable for growth and development of micro-flora is the beneficial effect of halogen treatment. Diminishes production of volatile aldehyde is the beneficial effect of halogen treatment, which is apparently the result of lipid peroxidation. Other than other impact, the anti-fungal role of chemical may give some helpful impact on germinability.

How to Halogenate the Seed?The seeds are exposed to very low concentration of any one of the following halogens like chlorine, bromine, iodine, alcohols, iso-propanol or ethanol in a closed airtight container for 16-72 hrs. The time of exposure and chemical concentration of exposure would depend on the initial quality of seed used for treatment. The halogens are acquired by adding 1ml of sulphuric acid to 50 mg of KCL or KBr or KI in a small petri-dish put in the base of a glass desiccators in which seeds are kept on the porcelain plate for required duration of exposure. (Methodology adapted from: Dharmalingam et al., 1998).



Methods for Application of Halogen Formulation to SeedApplication of fungicide seed treatment and pesticide seed similar to this the halogen formulation could be added to the seed. The chemicals compounds applied either dry dressing or slurry treatment utilizing 5 ml Kg-1 of seeds. The efficacy of slurry treatment is higher than dry treatment.

Halogenated Seeds in StorageThe shelf life of seeds prolonged during storage by treating the seeds with halogens like iodine or chlorine. Several research studies also reported that iodination is best for seeds to prolong their storability compared to chlorination.

Field Performance of Stored Halogenated SeedThe seeds treated with halogens in storage can likewise be prolonged to field level by increasing yield potential of the crop when compared to untreated seeds. Several researchers reported that the seeds treated with halogen can improve the seed yield to 7-15 percent.

Under laboratory condition post ageing germinability improved in rice also better field emergence and yield improvement due to halogen treatment.

Factors influencing halogen treatment1. Selectivity of treatment and crop2. Duration of treatment and dosage of halogen3. Dosage of halogen and age of the seed

Advantages of seed halogenations � Seed germination maintenance and protection

from pest during storage � Reduce the storage fungi � Rapid plant growth and high root intensity � Increased chlorophyll content and leaf area � Non-toxic and cost effective � Commonly easily available materials are used and

included in the seed production cycle

Disadvantages � Time consuming and laborious � When compared to other treatment the efficacy of

halogen treatment is low

PLANT PATHOLOGY

20115

65. Fusarium Wilt (Tropical Race 4): A Destructive Disease of Banana in IndiaAJIT KUMAR SAVANI 1*, K. DINESH2 AND PULI. SHASHANK ROY 3

1Department of Plant Pathology, Assam Agricultural University-785013 2 Department of Plant Pathology, University of Agricultural Sciences, Dharwad 3Department of Plant Pathology, Indira Gandhi Krishi Vidhya Peeth, Raipur

Introduction: Fusarium wilt of banana, also known as the Panama disease, is a historically important disease of bananas worldwide (Ploetz, 1990). The disease was first recognized in Australia in 1874 by Bancroft (Bancroft, 1876). Fusarium wilt epidemics in the twentieth century resulted in the devastation of more than 50,000 hectares of exotic Gros Michel (AAA) plantations (Ploetz and Pegg, 1997) leading to a major shift of the entire banana production to resistant Cavendish (AAA) banana varieties such as Williams, Grand Naine and Dwarf Cavendish. Recently a new malady for growers around the world is the global spread of Tropical Race 4 (TR4). The term TR4 was coined to distinguish this race 4 strain from the ones that affect cavendish cultivars only in the presence of predisposing factors such as low temperatures (Promusa, 2011). Since 2015 TR4 has spread to the states of Uttar Pradesh, Madhya Pradesh, Gujarat, Kathihar and Purnea districts of Bihar in India (Damodaran et al., 2018 and Thangavelu, 2019).

Symptoms

External symptoms � The external symptoms are expressed only 4-5

months after planting. However, if the diseased suckers are planted, the symptoms of the disease can be seen even 2 months after planting.

� At initial stage, yellowing of leaf margins of older leaves and later the yellowing progresses towards the midrib and finally the whole leaf become yellowish. Then the yellowing of leaves spreads to upper leaves as well.

� The infected leaves gradually collapse at the petiole or towards the base of the midrib and hang down around the pseudostem and this gives ‘skirt” like appearance to the plant

� The youngest leaves are the last to show the symptom and often stand erect giving ‘spiky’ appearance to the plant

� The newly emerging leaves will be pale with reduced leaf lamina and finally emergence of the leaf will be stopped.



� Longitudinal splitting of the pseudostem and emergence of large number of side suckers before the death of infected plants.

� Normally bunches are not produced and if produced, the fruits are small with few developed fingers.

Internal symptoms � Presence of yellow, red or brown strands on

the corm and continuous black or brown or yellow coloured strands in the pseudostem and sometimes also in the bunch stalk due to the discoloration of vascular tissues caused by the fungus.

Integrated Disease Management of Fusarium Wilt of BananaThe disease can be managed effectively only by following integrated disease management (IDM) practices. They are

� Keep the sign board (indicating beware of TR4 with danger sign and restricted entry) at the entry of each field affected with Fusarium wilt Tropical race 4.

� Demarcate the wilt infected plants with rope/ coloured ribbon for restricted entry inside the field.

� Inject the wilt infected plants with Glyphosate 2-5 ml/ plant in two different places (preferably one at the bottom and second 2 feet above from the ground).

� Follow “come clean and go clean” approaches (wear polythene shoe or foot cover while entering into the field and the same may be removed before the exit of the field. The same may be preserved for next use). Also 2 plastic drums with tap connection at the bottom of the drum may be kept at the entrance of the field. One for keeping water and another one for storing disinfectant (1% poly dimethyl ammonium chloride @ 10 g in 1 lire of water). All the tools used including hand and foot may be washed first in water and latter in

disinfectant � Follow the good agricultural practices so as

to improvement of soil health by applying recommended dose of fertilizers (apply less of N and more of KO, prefer only nitrate nitrogen), apply 1 kg of wood ash, more 2 amount of organic manures such as vermicompost, neem cake, well decomposed farm yard manure etc, (for this banana waste recycling may be followed) application of effective microbes etc

� As soon as the sign of wilt infection is noticed, drenching of Carbendazim (0.1 to 0. 3%) @ 3-5 litres per plant for 3-5 times at 15 days interval and pseudostem injection of 3 ml of 0.1% carbendazim solution at 3rd 5th and 7 th month after planting for all the plants (both Infected and non-infected) may be carried out

ReferencesBancroft, J. (1876). Report of board appointed to

inquire into the cause of disease affecting livestock and plants. In: Votes and proceedings. pp. 1011-1038

Damodaran, T., Mishra, V. K., Jha, S. K., Gopal, R., Rajan, S., & Ahmed, I. (2018). First Report of Fusarium wilt in banana caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in India. Plant Disease. PDIS-07.

Promusa. (2011). Tropical race 4. Promusa secretariat. Biodiversity international. Department of agriculture. France. 540p.

Ploetz, R.C. (1990). Variability in Fusarium oxysporum f. sp. cubense. Canadian Journal of Botany. 68: 1357-1363

Ploetz, R.C. and Pegg, K.G. (1997). Fusarium wilt of banana and Wallace’s line: was the disease originally restricted to his Indo-Malayan region? Australasian Plant Pathology. 26(4): 239-249.

Thangavelu, R., Mostert, D., Gopi, M., Devi, P. G., Padmanaban, B., Molina, A. B., and Viljoen, A. (2019). First detection of Fusarium oxysporum f. sp. cubense tropical race 4 (TR4) on Cavendish banana in India. European Journal of Plant Pathology.1-10.

20118

66. Life Cycle of PythiumASHUTOSH C. PATIL1 AND ANANTA G. MAHALE 2

1 Ph.D. Scholar, Department of Plant Pathology, VNMKV, Parbhani. 2 Ph.D. Scholar, Division of Soil Science & Agriculture Chemistry, SKUAST-Kashmir, Srinagar.

Taxonomic Position of Pythium Domain: EukaryaKingdom: ChromistaPhylum: OomycotaClass: OomycetesOrder: PythialesFamily: Pythiceae

Taxonomic Position of Pythium Genus: PythiumSpecies: aphanidermatum



Symptoms

Pre-emergence damping-off � Common in nursery. � Rotting of seed and radicle before the seedlings

emerge out of the soil.

Post-emergence damping-off � Anew emerged seedlings are killed at ground level

after they emerge from the soil causing them to collapse.

� Pectolytic and cellulolytic enzymes produced by Pythium plays an important role in tissue rupture and debilitation.

Perpetuation � Primary infection is through seed and infected

soil. � Secondary infection through Zoospores carried by

irrigation water.

Life Cycle

Asexual reproduction � The mycelium consists of slender, coenocytic

hyphae with cellulose walls; hyphae are both intracellular and intercellular.

� Haustoria are absent. � They produces filamentous or simple or branched,

lobed or globose zoosporangia terminally or intercalary on the rapidly growing hyaline mycelium.

Life cycle of Pythium sp.

Germination is either by zoospores or germ tube. The sporangiophore consists of sporangia with a bubble like vesicle. The sporangial protoplast moves rapidly through the tube into the vesicle and seem to remain mute while the demarcation of the zoospores is taking place. The zoospores when grow ripe exhibit rocking effort and bounce on the vesicle wall, The vesicular wall rupture like a soap bubble and the zoospores disperse in all directions. The reniform zoospore has two lateral flagella attached to the concave side. The tinsel flagellum is directed anteriorly and the whiplash

projects posteriorly when the zoospore swims in the film of water. Later a period of swimming, the spore comes to rest, encyst and germinates by producing a germ tube.

Sexual reproductionSexual reproduction is oogamous (gametangial contact). The male sex organ (anthredium) and the female (oogonium). The oogonium is globose with an outer layer of periplasm and inner ooplasm (oosphere/egg). Antheridia (paragynous) are much small and



somewhat elongated/club shaped arising laterally either from the oogonial stalk or from the neighbouring hypha. Farther gametangial contact, a fertilization tube develops and penetrates the oogonial wall and enters the periplasm. In the meanwhile meiosis takes place in both the gametangia. Only one functional nucleus remains, others disorganise. Male nucleus now passes through the tube into the oosphere, approaches the female nucleus, unites with it and forms the zygote. The

oosphere develops into a thick smooth walled aplerotic oospore, which is differentiated into two layers viz., the outer exine and inner intine.

� P. aphanidermatum and Pythium myriotylum are the two species mostly severe at higher temperatures.

� P. irregulare and Pythium ultimum are the two species mostly severe in moist and cool conditions

20119

67. Metabolome of Trichoderma: A Novel Strategy for Plant Defense and GrowthPULI SASANKA ROY1*, AJIT KUMAR. SAVANI 2 AND K. DINESH3

1Department of Plant Pathology, Indira Gandhi Krishi Viswavidhyalaya, Raipur (C.G.), 2Department of Plant Pathology, Assam Agricultural University- Jorhat, Assam, 3Department of Plant Pathology, UAS, Dharwad, Karnataka.

IntroductionDue to many recent developments in modern agriculture, we are depending on agrochemicals which degrade the soil leading to environmental pollution. Hence, several studies have been done for application of microorganisms for plant growth and biocontrol of diseases. Among them, Trichoderma spp. have been studied widely, and are presently marketed as biopesticides and biofertilizers, due to their ability to protect plants, promote vegetative growth and control various phytopathogenic agents under different agricultural conditions. The important factor that contributes to their beneficial biological properties is the wide variety of secondary metabolites they produce. The genome sequences of Trichoderma spp. have revealed a vast repository of genes putatively involved in production of secondary metabolites (SMs) (Mukherjee PK et al., 2012). Theses metabolites were found to inhibit the growth and pathogenic activities of the parasites directly and also increase disease resistance by triggering the defence system in the plants. Moreover, these metabolites are capable of enhancing plant growth that enables the plant to counteract the diseases with compensatory vegetative growth and increases nutrient uptake from the soil. The benefits of the Trichoderma can be summarized as:

Benefits1. Rhizosphere competence, allows rapid

establishment of a stable microbial community within the rhizosphere.

2. Suppression of the pathogens using a variety of mechanisms.

3. Overall improvement of the plant growth.4. Plant growth promoters.5. Enhances nutrient availability and uptake.6. Induction of host resistance similar to that

stimulated by beneficial rhizobacteria (Harman G.E. et al., 2004).The production of SMs from Trichoderma

depends upon the strain and varies in relation to the equilibrium between elicited biosynthesis and biotransformation rates. It was reported that Trichoderma produces SMs more than 100 different structures which includes low molecular mass-non- polar compounds such as pyrones, terpenoids, steroids, polyketides, peptaibiotics, NRPSs, siderophores etc. The production of these SMs are regulated by environmental factors, growth substrate etc.

T. virens (Hypocera virens) and T. atroviride (Hypocera atroviridis) are aggressive mycoparasites having diverse repertoire of biosynthetic clusters as large compounds which are necessary to attack against other microbes. Ghisalberti and Sivasitamparam had classified these metabolites into three categories: i) volatile antibiotics, i.e., 6-pentyl-α-pyrone (6PP) amd most of the isocyanine derivatives; ii) water soluble compounds, i.e., heptelidic acid or koningic acid; and iii) peptaboils, linear oligopeptides of 12-22 amino acids rich in α-aminoisobutyric acid, N-acetylated at the N terminus and containing an amino alcohol (pheol or Trpol) at the C terminus.

Sms in Plant Defence and SignallingSMs may act as either elicitors or resistance inducers during the interaction Trichoderma with the plants (Harman et al., 2004). They may be i) proteins with enzymatic activity (Xylanase), ii) avirulence gene like products that induce defence reactions in plants and iii) low molecular weight compounds released from fungal or plant cell walls by the activity of Trichoderma enzymes. They can cause an overall increased production of defence related enzymes like peroxidases, chitinases, β-1-3, glucanases and lipoxygenases (Mukherjee PK et al., 2012).



Gliotoxin was the first metabolite isolated and described from Trichoderma (Brain, 1944). It is a fungistatic metabolite and belongs to ETPs (epipolythiodioxopiperazines) class of peptides. It was implicated in antagonism against Rhizoctonia in the soil (Weindling and Emerson, 1936). The compound is profusely found in rhizosphere of the soil and has received much attention due to its role in biocontrol of soil borne pathogens. Gliovirin is another ETP compound described from Trichoderma. It has high potent antimicrobial properties especially against oomycetes (Howell et al., 1993).

Peptaibols largely produced by Trichoderma/Hypocera are ecologically and commercially important for antimicrobial and anti-cancer properties as well as their ability to induce systemic resistance in plants against microbial invasion. The ability of peptaibols to self-assemble forming voltage dependent ion channels in membranes is responsible for antibiotic properties. Alamethicin and Trichovirin II are prominent among these compounds. The application of Alamethicin produced by T. viride is shown to induce local and long distant-electric signals in plants similar to hypersensitive responses against pathogen attack (Maischak et al., 2010). The application of Trichovirin II produced by T. virens has shown increased systemic resistance response and produced higher levels of phenolic compounds in cucumber plants (Viterbo et al., 2007).

Pyrones (6-pentyl pyrone, 6PP), the ‘coconut aroma’ volatile compound produced by Trichoderma spp. is one of the best described SMs in a biocontrol prespective. These compounds are known for its anti-fungal properties and plant growth promoting activities (Vinale et al., 2008).

Sms in Plant GrowthTrichoderma spp. produces organic acids such as gluconic acids and fumaric acids that decreases soil pH and allow the solubilisation of phosphates, micronutrients and mineral cations like iron manganese, magnesium, useful for plant metabolism. These compounds help plants to absorb nutrients in alkaline or neutral soils.

Trichoderma ssp. has ability to enhance plant biomass production, promoting lateral growth through auxin dependent mechanism (IAA). T. koningii produces koninginins (A-C, E, G;) that acts as plant growth regulators and had a concentration dependent activity in a wheat coleoptile assay (Cutler et al., 1989).

Microorganisms have a high-affinity iron system based on Fe3+-chelating molecules, called siderophores. Siderophore production by microorganisms can be beneficial to plants for two reasons; i) can solubilize iron unavailable for plants, ii) siderophores produced by beneficial microorganisms can also suppresses the growth of pathogens by depriving iron in them (Leong J, 1986).

ConclusionSeveral studies indicated that numerous metabolic

changes occur in the roots of the plant during colonization by Trichoderma ssp. i.e., activation of pathogenesis related proteins and increase the induced resistance to subsequent attack by numerous microbial pathogens of all classes (fungi, bacteria and oomycetes). The isolation of these metabolome will help to overcome the problems related to pathogens and promote plant growth in unfavourable conditions. The metabolome can be produced inexpensively in large scale and can be separated from the fungal biomass easily. No harmful effects are observed in humans due to these metabolites and so eco-friendly. The application of Trichoderma metabolome to induce host resistance and promote plant growth may come to true in near future.

ReferencesCutler HG, Himmetsbach DS, Arrendale RF, Cole

PD, Cox RH. (1989) Koninginin A: a novel plant regulator from Trichoderma koningii. Agricultural and Biological Chemistry, 53, 2605-2611.

Harman GE, Howell CR, Viterbo A, Chet I, Lorito M. (2004) Trichoderma species - opportunistic, avirulent plant symbionts. Nature Review Microbiology, 2, 43-56.

Howell, C. R., Stipanovic, R. & Lumsden, R. (1993). Antibiotic production by strains of Gliocladium virens and its relation to biocontrol of cotton seedling diseases. Biocontrol Sci Technol 3, 435–441.

Leong J. (1986) Siderophores: their biochemistry and possible role in the biocontrol of plant pathogens. Annual Review of Phytopathology, 24, 187–209.

Maischak, H., Zimmermann, M. R., Felle, H. H., Boland, W. & Mitho¨ fer, A. (2010). Alamethicin-induced electrical long distance signaling in plants. Plant Signal Behav 5, 988–990.

Mukherjee PK, Horwitz BA, Kenerley CM. (2012) Secondary metabolism in Trichoderma – a genomic perspective. Microbiology, 158, 35–45.

Sivasithamparam, K. & Ghisalberti, E. (1998). Secondary metabolism in Trichoderma and Gliocladium. In Trichoderma and Gliocladium Basic Biology, Taxonomy and Genetics, pp. 139–191. Edited by C. Kubicek & G. E. Harman. London: Taylor & Francis.

Vinale, F., Sivasithamparam, K., Ghisalberti, E., Marra, R., Woo, S. & Lorito, M. (2008). Trichoderma–plant–pathogen interactions. Soil Biol Biochem 40, 1–10.

Viterbo A, Wiest A, Brotman Y, Chet I, Kenerley C. (2007) The 18mer peptaibols from Trichoderma virens elicit plant defence responses. Molecular Plant Pathology, 8, 737-746.

Weindling, R. & Emerson, O. (1936). The isolation of a toxic substance from the culture filtrate of Trichoderma. Phytopathology 26, 1068–1070.



20124

68. Plant Metabolite Engineering / Manipulation for Plant DefenceBANDANA SAIKIA*1, S. AJIT KUMAR1 AND K. DINESH2

1Department of Plant Pathology, Assam Agricultural University, Jorhat-785013 2Department of Plant Pathology, University of Agricultural Sciences, Dharwad- 580005

IntroductionPlants produce a myriad of secondary metabolites with an immense number of applications. These compounds are important for their interaction with the environment and their survival such as in, symbiosis, as pollinator attractants and defence against herbivores and phytopathogens. Biotic and abiotic factors that limit crop productivity are defended by plant through different mechanisms. Plant secondary metabolites, including terpenes, phenolics and nitrogen (N) and sulphur (S) containing compounds, are the key participant of defence reactions against a variety of herbivores and pathogenic microorganisms as well as various kinds of abiotic stresses. Manipulation of environmental plant stress factors to stimulate biosynthesis of metabolites to tilt plant metabolism towards desired output is an environment friendly way to ensure food security for the growing population.

Secondary Metabolites in Plant DefenceSecondary metabolites (terpenes, phenols, sulphur and nitrogen substances) unlike the primary metabolites donot have direct role in growth and development but high concentration might result in a more resistant plant. Defence metabolites when present as constitutive substances are called prohibitins or phytoanticipins and when synthesized in response to an infection involving de novo enzyme synthesis, known as phytoalexins. Phytoanticipins are high energy and carbon consuming and exhibit fitness cost under natural conditions, but recognized as the first line of chemical defence that potential pathogens have to overcome. In contrast, phytoalexin production may take two or three days, after infection.

Metabolite Engineering in Plant DefenceMetabolic engineering is a process for modulating the complex plant metabolic pathways or the metabolism of the organisms so as to produce the required amounts of the desired metabolite with appreciation of environmental influence. This can be achieved through genetic manipulation or by manipulation of environmental plant stress factors to stimulate biosynthesis of metabolites. Process of genetic manipulation is expensive and requires time and skilled personal. Also, inadequate public acceptance and soil specificity of genetically modified food are still the challenges. Hence, manipulation of plant environment to synthesize defence metabolite is the

way that reaches the farmers.

Manipulation of Plant EnvironmentApplication of Plant growth promoting microorganisms (PGPM) including rhizospheric and/or endophytic microbes has been found to improve plant health. Unlike microbial biopesticides, PGP microorganisms do not necessarily inhibit the pathogens by the mechanism of competition, antibiosis or enzymatic lysis but they may have these characters. Microbes inhabiting the plants not only stimulate plant growth through nutrient acquisition but also provide fundamental support in tolerating biotic stresses directly by microbial secondary metabolites and indirectly by stimulating plant metabolism (Shahid and Mehnaz, 2020). Another approach to manipulate host environment is by application of micro nutrients. In a series of experiments Massey and Hartley (2006, 2007 and 2009) showed that application of silicon to soil is inversely proportional with rice blast severity limiting occurance of the disease by 10%. Grasses take up silicon from the soil in unusually high amounts and deposit it in their leaves making it difficult for blast pathogens to enter the host. In higher amount deposited silicon form sharp granules called phytoliths which make leaves more abrasive to herbivore mouthparts.

ConclusionMicrobe mediated stress tolerance to plants have gain the limelight in past two decades. Many of these microbes have been commercialized. However, their field application id hampered due to lack of knowledge on interrelated metabolic pathways and chemical compounds to suppress pathogens. To overcome this problem, potent microbes should be studied in crops. This may open the doors to harness the potential benefits of plant based biochemicals to make agriculture more sustainable and less reliant on chemical inputs.

ReferencesMassey, F.P and Hartley, S.E. (2006). Experimental

demonstration of the anti-herbivore effects of silica in grasses: impacts on foliage digestibility and vole growth rates.

Massey, F.P and Hartley, S.E. (2009). Physical defences wear you down: progressive and irreversible impacts of silica on insect herbivores. Journal of Animal Ecology 78: 281-291

Massey, F.P. Ennos, R. and Hartley S.E. (2007).



Herbivore specific induction of silica-based plant defences. Oecologia 152: 677-683

Shahid, I and Mehnaz, S. (2020). Microbial secondary

metabolites: Effectual Armors to Improve Stress Survivilaty in Crop Plants. Microbial services in Restoration Ecology, 47.

20130

69. Ontogenic ResistanceP. AVINASH

Division of Plant Pathology, PhD Research Scholar, SKUAST-Kashmir.

Disease resistance mechanism helps the plants to protect by reducing the growth and development of various pathogens. Plant resistance is of 3 types they are true resistance, non-host resistance and apparent resistance.

Ontogenic resistance is a different approach of resistance. The term ‘Ontogeny’ is derived from two Greek words ‘ontos’ and ‘geny’ means developmental history of an organism within its own life. “It describes the ability of whole plants or plant parts to resist or tolerate disease severity as they age and mature”. The effect of plant age on resistance was first described by Anderson et al., 1947 in tomato against leaf mould disease caused by Cladosporium fulvum.

In ontogenic resistance, the ability of whole plants or plant parts to resist or tolerate disease as they age and mature. It may not lead to immunity, but levels of resistance that develop during aging of plant tissue may greatly affect eventual disease severity and may even lead to escape from infection. For example, bean seedlings are highly susceptible to damping off, caused by Pythium ultimum, during the first 6 days after emergence. Later, infection may still lead to some root necrosis but will not result in the collapse of the entire plant.

Forms of Ontogenic Resistance

1. Resistance and developmental transitionsIt may be developed gradually during life of the plant associated with major transitions of plant life cycle. It may be established at Adult transition.

Eg: Tobacco black shank caused by Phytophthora parasitica the resistance correlated with time of flowering stage.

2. Resistance and tissue maturitySeveral plant species develop resistance that is restricted to a given tissue or organ as a function of the maturity of that tissue or organ.

Eg: Soybean root rot caused by Phytophthora sojae in hypocotyl the resistance varies with tissue maturity.

3. Increased or acquired resistance and plant developmentAs the plants become develop and mature, they show

the increased or acquire resistance for the pathogens.Eg: In tobacco to TMV and Peranospora tabacina.

(tobacco blue mold disease)

4. Specific and broad-spectrum resistanceOntogenic resistance may be effective against several pathogens particular race

Eg: specificIn bacterial blight of rice caused X. oryzae pv. oryzae, is developmentally controlled by Xa21 R gene.

Eg: broad spectrumTobacco against Peranospora tabacina (blue mold), Phytophthora tabacina (root rot) expresses resistance during flowering stage.

Impact on Control � Precise definition of onset of age, age related

resistance (ARR). � Optimization of fungicide application schedules

accordingly. � Understanding of the mechanisms and impact on

growth of pathogen helps to predict epidemics. � ARR or ontogenic resistance represents an

additional tool that can be exploited to better management of plant disease.Molecular mechanisms for development of

defense mechanisms are through regulation of R genes, Pathogenesis Related protein (PR) synthesis and salicylic acid pathway. In many pathosystems, ontogenic resistance is quite commonly observed and noted. It is less commonly integrated in disease management systems, though. A fundamental principle of integrated pest management (IPM) is the alignment of control measures with the actual risk of infection and potential for loss. It seems that knowledge of ontogenic resistance is an underutilized resource in devising IPM strategies that could be used to improve plant disease management.



20176

70. Post-Harvest Diseases Caused by Abiotic FactorsP. VALARMATHI

ICAR- Central Institute for Cotton Research (CICR), Coimbatore

Postharvest LossesMost losses of fresh produce occur between leaving the farm and reaching the consumer. Losses during this period have been estimated to be about 20% of the total crop. These losses may be caused by complete wastage of the product or by lower prices due to a reduction in quality. There are generally three main causes of postharvest losses: 1. Disease caused by fungi and/or bacteria, 2. Physical injuries due to insects, mechanical force, chemicals, heat or freezing and 3. Non-disease disorders resulting from storage conditions that upset normal metabolism

Postharvest DisordersDisorders are the results of stresses related to excessive heat, cold, or improper mixtures of environmental gases such as oxygen, carbon dioxide, and ethylene. The term “disorder” refers to those problems in fruit not caused by pathogens. Symptoms are induced by fruit reacting to some kind of stress connected with temperature, light, humidity, atmosphere, or handling. SOME disorders may be caused by mechanical damage, but all are abiotic in origin (not caused by disease organisms) and cannot be controlled by chlorination or most other postharvest chemicals. However, abiotic disorders often weaken the natural defenses of fresh produce, making it more susceptible to biotic diseases those that are caused by disease organisms. Further, in many cases injuries caused by chilling, bruising, sunburn, senescence, poor nutrition, and other factors can mimic biotic diseases.

Physiological DisordersIt can be divided into five general categories: nutritional, temperature-related (low and high), respiratory, senescent and miscellaneous. Nutritional and low temperature disorders are particularly problematic and

are considered in detail. Sunburn on the shoulders of fruit, such as tomato and mango, is a common example of high temperature injury incurred prior to harvest. Respiratory disorders are associated with low oxygen and or high carbon dioxide concentrations in and around harvested produce in controlled atmosphere storage and modified atmosphere packaging. Black heart of potato is an example of low oxygen injury and midrib browning of lettuce is an example of carbon dioxide injury. Senescence disorders, such as mealiness in apples, are generally associated with harvesting over-mature produce and or over storing produce. Other miscellaneous disorders tend to be cause and or product-specific in terms of the relatively unique symptoms expressed. Example, exposure to ethylene causes russet spotting on the midrib of lettuce leaves and bitterness (isocoumarin accumulation) in carrots. Greening of potatoes exposed to light and rooting of onions exposed to high humidity may also be considered miscellaneous physiological disorders.

The cellular biochemical and biophysical mechanisms that give rise to physiological disorders in produce are extremely complex. Moreover, they often involve elusive interactions with the pre- and post-harvest conditions. In diagnosing causes of physiological disorders or attempting to predict the likelihood of symptom expression in harvested produce, the following parameters may need to be taken into account: preharvest environment conditions (eg., temperature, nutrition and water regimes), crop development factors (eg. Yield or crop load, position on the plant and carbohydrate, water and or nutrient partitioning) and postharvest environment conditions (eg., temperature regime, gas atmosphere and storage time).



DISEASE MANAGEMENT

20168

71. Management of Rice DiseasesPRINCE KUMAR GUPTA*, SADHNA CHAUHAN, MANOJ KUMAR CHITARA AND SNEHA SHIKHA

Ph.D. Scholar, Department of Plant Pathology, College of Agriculture, GBPUAT, Pantnagar-263145 *Corresponding Author Email: [email protected]

IntroductionRice (Oryza sativa L.) being the most essential cereal is consumed by about 50% of the world population. According to FAO, it is considered as a staple food providing 2400 calories/ day that is the least food safety required a person/ day. In 2004, “Rice is Life” the theme of International Year of Rice that reflected the importance of rice, which hold the key to our country’s ability to produce enough food for our

people. India is the second-largest rice-producing country by contributing production of 112.91(MT) in area of 43.79 (000ha) with yield 2578 kg/ha after china (DES, 2018). In India, west Bengal is the leading state in rice production followed by U.P., Punjab, T. Nadu, Andra Pradesh and Bihar. Unfortunately, this crop is susceptible to several diseases which reduced its economic importance. So different diseases caused by a pathogen (Fungus, Bacteria and Viruses) are listed in table 1.

Table. 1. Different diseases of rice and their management

Important disease & it’s causing agent

Symptoms Managements

Fungal diseasesRice blast(Pyricularia grisea)

The fungus invades nearly all above-ground parts of the crop. Depending on the site, rice blast symptoms are-Leaf blast, an elliptical or spindle-shaped lesion with brown borders having grey centres is produced which under favourable conditions become enlarge and coalesce which eventually kills the leaves.Collar blast appears when the pathogen infects the collar which later kills the whole leaf blade of the crop. The node of the stem starts turning into blackish and can breaks easily, the condition called a node blast. Infected neck (neck blast) symptoms showed girdling by a greyish brown lesion which leads panicle to fall over the severe infection. The formations of brown lesions on the branches of panicles and the spikelets are also produced by a fungus.

Reduced application of nitrogenous fertilizer.In endemic areas, adoption of seed treatment with Tricyclazole 75 WP @ 2 g/kg or Carbendazim 50 WP @ 1 g/kg found effective in disease control.Foliar spray Tricyclazole 75 @ 0.6g/litre or Carpropamid 30 SC @ 1ml/litre. Or Isoprothiolane 40 EC @ 1.5 ml/litre or Iprobenphos 48 EC @ 2ml/litre or Kasugamycin-B 3 [email protected] ml/litre or Carbendazim 50 WP @ 1 g/litre reduced the incidence of disease.4. Growing of blast resistant varieties like Rasi, IR 64, Prasanna, IR 36, Vikas, Tulasi, Sasyasree etc.

Sheath blight(Rhizoctonia solani)

Symptoms initiate with lesions formation on the leaf sheaths although leaf blades may also be affected. Normally, small, ellipsoid or ovoid lesions of greenish-grey colours are developing near the water line in lowland fields. Under favourable conditions, lesions become enlarge and coalesce results in the formation of bigger lesions having an irregular outline and greyish-white centre with dark brown borders. On plant leaf sheath the presence of numerous large spots causes the death of the whole leaf.

Spray of Validamycin 3 L @ 2.5ml/litre or Thifluzamide 24 SC@ 0.75 g/litre or Hexaconazole 5EC @ 2 ml/litre or Propiconazole 25 EC @ 1ml/litre or Carbendazim 50 WP @ 1g/litre found effective.2. Apply 2-3 split application of nitrogen fertilizer.





Brown spot(Bipolaris oryzae)

Brown spot may also appear as seedling blight, foliar and glume disease. On seedlings, a small, circular and brown lesions are produced which may girdle the coleoptile and leads to distortion of the primary as well as secondary leaves. The root discolourations of the infected plant are also produced on an infection. On the leaves of older plants, a circular to oval lesions having light brown to grey centre which is surrounded by a reddish-brown margin are produced by the fungus. Further severity causes coalition of the lesion which results in the killing of large areas of affected leaves. The fungus may also infect the glumes producing dark brown to black oval spots, and later the grain also gets infected and leads to black discolouration

Proper crop nutrition with clean cultivation.Avoidance of water stress.Treatment of seed with Mancozeb (63%) + Carbendazim (125) @ 2g/kg of seed is effective in the control of the disease.Resistance varieties such as Rasi, IR36, Jagannath should be grown.

Sheath Rot (Sarocladium oryzae)

Normally, rotting will occur on the leaf sheath which encloses the young panicles. The oblong or somewhat irregular spots lesion of 0.5-1.5 cm long, with grey to light brown centres that are surrounded by dark reddish-brown margins is formed. On later, lesions enlarge and coalesce which cover major parts of the leaf sheath. Lesions may also concur sow reddish-brown discolouration in the sheath. Profuse whitish powdery growth may also found inside the affected sheath. The panicle may fail to emerge completely at severe infection so the young panicles remain within the sheath or only partially will emerge. Non-emerged panicles tend to rot, which turns florets red-brown to dark brown.

Seed treatment with Mancozeb 75 WP @ 2.5 g/kg or Captan 50 WP will be effective in disease control.Spraying of Mancozeb 75 WP @ 2.5 g/kg or Hexaconazole 5 EC @ 2 ml/litre or Propiconazole 25 EC @ 1 ml/litre or or Thiophanatemethyl 70 WP @ 1 g/litre effective in field to control the disease progress.

False Smut (Ustilaginoidea virens)

The disease occurs at hard dough to mature stage of the crop. The individual grains of the panicle get transformed into greenish spore balls having velvety appearance. Initially, the spore balls are small and then visible in between glumes, which gradually grow to reach 1 cm or more in diameter and covers the floral parts. At further growth, the membrane gets burst and release spore balls. Later on, the ball becomes orange and yellowish-green or greenish-black. The ball surface gets crack at this stage. The outermost layer of the ball is green and consists of mature spores together with fragments of mycelium. The outer soporiferous area has three-layered. The outermost layer is greenish-black having powdery spores; the middle layer is orange while the innermost layer is yellowish.

Use of disease-free seed for sowing.Hot water treatment of seed @ 52ºC for 10 min Spray was effective in control.Removal of diseased panicles in the field.Seed treatment with Carbendazim @2.00g/kg of seed.Spray of Propiconazole 25 EC @ 1 ml/litre or Chlorothalonil 75 WP @ 2 ml/litre or Copper oxychloride at around flowering.

Bacterial diseasesBacterial Blight (Xanthomonas oryzae pv. Oryzae)

Usually, water-soaked lesion starting at leaf margins, a few cms from the tip, and spread to the leaf base; affected areas become yellowish to light brown due to drying; with the yellowish border between the dead and green portion of the leaf which usually observed at the maximum tillering stage and onwards. In severe infection, it causes withering of leaves or entire young plants (refers as kresek) and production of pale-yellow leaves at a later stage of the growth which also reduced grains quality.

Application of judicious amount of fertilizer (60-80 kg N and required dose of K), Apply N in 3-4 split.Destroyed infected stubbles and weed.Use of resistance varieties like Ajaya, IR64, IR-20 etc.4. Spray 0.05g Streptocycline and Copper Sulfate.





Bacterial Leaf Streak(Xanthomonas oryzae pv. Oryzicola)

A symptom first appears as short, water-soaked streaks between the veins, which later become longer and translucent and turn to light brown or yellowish-brown. Due to numerous streaks large leaf area may become dry. At the late stage, the disease is indistinguishable from the BLB.

Field sanitation by removal of weed host and stubbles.Use balanced N fertilizers.Use of resistant varieties.Use of copper-based fungicides at heading stage.

Viral diseasesTungro(Rice tungro bacillifor virus and spherical virus)

Infected plants become stunted. Later leaves become orange-yellow or yellow. The leaf discolouration starts initially from the tip and spread to the lower part of the leaf blade; often only the upper portion is discoloured. Young leaves may give mottled appearance while old leaf gives rusty-coloured specks of various sizes. The panicles are small and not completely exerted, and bear mostly sterile or partially-filled grains often covered with dark brown specks. Infections also result in a delay in flowering. Disease transmits by green leafhoppers.

Grow resistance varieties such as IR-36/MTU-9002, 1003.Removal of the diseased plant from the field.In nursery apply Carbofuran 3G @10kg/ac or Phorate 10 G 2 5kg/ac found effective in control.Spraying of Monocrotophose 36EC @400ml/ac or Cabaryl 50WP @600g/ac etc.

Grassy Stunt (Rice grassy stunt virus)

Infected plants show severe stunting; excessive tillering, with short leaves that are narrow and, pale green to pale yellow. The plant may have newly-expanded leaves that might be mottled or striped and may also have numerous small, irregular, dark brown or rust-coloured spots. Brown planthopper is a carrier of the virus.

Removal of the infected plant from the field.Grow resistance varieties viz; Nidhi, Radha, Triveni etc.Use of Carbofuran 3G @10g/ac or Isazophos 3G @10g/Ac in a nursery.Spray monocrotophose 36EC @400ml/ac.

Ragged Stunt (Rice ragged stunt virus)

Stunting with reduced tillering is an initial symptom of the disease. The infected leaves are short, dark green, and serrated along one or both edges giving a ragged appearance. The leaf blades become twisted and form a spiral. The vein swellings also appear on leaf sheaths, leaf blade and culms and nodal branches are developed at later growth stages. Brown planthopper transmits the disease.

Removal of the infected plant from the field.Grow resistance varieties viz; Nidhi, Radha, Triveni etc.Use of Carbofuran 3G @10g/ac or Isazophos 3G @10g/ac in a nursery.Spray monocrotophose 36EC @400ml/ac.

References Agricultural Statistics at a Glance. Directorate of

Economics & Statistics. Department of Agriculture, Cooperation & Farmers Welfare. Ministry of Agriculture & Farmers Welfare. Government of India. Krishi Bhawan, 2018; New Delhi-110 00.

Agrios, G.N. (2005). Plant Pathology. Florida: Elsevier academic press.

Food and Agriculture Organisation (FAO), 2000. Agriculture towards 2015/30. Technical Interim Report. April 2000 Rome.

Singh, R.S. (2009). Plant Disease. Oxford and IHB Publication.

NEMATOLOGY

20112

72. Horizontal Gene Transfer is a Blessing for Nematode Parasitism Towards PlantsPRANAYA PRADHAN1* AND JYOTI PRAKASH SAHOO2

Ph.D. Research Scholar, 1Department of Nematology, 2Department of Agricultural Biotechnology, College of Agriculture, OUAT, Bhubaneswar – 751003 *Corresponding Author Email: [email protected]

Nematodes are estimated to include more than one million species, and are present worldwide in marine, freshwater and terrestrial habitats. The most well-

known representative is Caenorhabditis elegans; the first animal of which the genome was completely sequenced. C. elegans is a bacterivorous soil-inhabiting



nematode, and other members of the species phylum Nematoda show a wide range of trophic ecologies. Apart from bacterial feeders, fungivorous nematodes, plant parasites, predators, omnivores, and parasites of invertebrates and vertebrate animals, including humans, are represented. Among plant parasitic nematodes, cyst (Globodera and Heterodera spp.) and root knot (Meloidogyne spp.) nematodes are most notorious for causing major damage to crops such as soybean, potato, and sugarbeet. So here comes the gene transfer regarding the nematode parasitism says that the transmission of genetic information in the form of DNA from one cell to another cell. It is the basis for evolution.

So there is two types of Gene transfer:

A. Vertical Gene Transfer: Transmission of genes from an organism to its offspring Eg-Sexual reproduction in higher animals and plants is the way of vertical gene transfer.

B. Horizontal gene transfer: (HGT) implies the non-sexual exchange of genetic material between species, in some cases even across kingdoms.

Horizontal gene transfer is of two types based on the time aspects.i) Ancient: Genes transferred between

organisms separated a long time ago. The ancient HGT is difficult to detect through codon usage bias and differential base composition.

ii) Recent: Genes transferred between organisms separated recently. Easy to detect based on Criteria of codon usage bias and differential base composition. (Luis Boto., 2009).

Horizontal gene transfer is goes on the three processs- Transformation, Conjugation, Transduction. It has been hypothesized that the ability of nematodes to parasitize plants was acquired by HGT from soil bacteria to bacterivorous nematodes. Nematodes as a group are suitable to study this hypothesis because within the phylum Nematoda plant parasitism arose independently multiple times, and because this feeding type is associated with a number of known, relatively well characterized genes. In this chapter we concentrate on the origin of cellulases, enzymes that are used by plant parasitic nematodes to depolymerise plant cell walls and the functions or actual benifits of HGT in nematodes parasitism.

Origin and Distribution of Horizontally acquired Genes in PPNIn 1998, Smant and co-workers discovered that the potato and soybean cyst nematodes (G. rostochiensis and H. glycines produce and secrete β-1,4-endoglucanase (cellulases). This finding constituted the starting point of a series of papers reporting a range of plant cell wall degrading enzymes (CWDE) from plant parasitic nematodes, including pectate lyases, exo polygalacturonase, endo-ß-1,3-glucanase, endoxylanases, expansins and cellulose binding proteins. Phylogenetic analyses suggest that

plant parasitic nematodes arose from fungivorous ancestors, which had evolved from bacterivores. In 1998, according to Keen and Roberts the nematodes borrowed the cellulase genes from microorganisms at some point in their evolution. This hypothesis is exciting, and at the same time hard to prove.

Cellulases are a good object to focus on when investigating HGT in phytopathogens, because they are widely produced by pathogens that attack plants (including bacteria and fungi) but not found in pathogens that attack animals. Cellulases are produced by many plant pathogenic and plant-feeding organisms and are a rather diverse group of enzymes as illustrated by the fact that cellulases can be found in 14 different glycoside hydrolase families (GHF 5, 6, 7, 8, 9, 10, 12, 26, 44, 45, 48, 51, 61, 74). The cellulases that have been identified and characterized from plant parasitic Tylenchida (Meloidogyne, Heterodera and Globodera and migratory parasitic species Pratylenchus penetrans) are from glycoside hydrolase family 5 (GHF5). Since the discovery of the first GHF5 nematode cellulase in 1998, there has been a strong over representation of bacterial cellulases in the GHF5 family. While cellulases from other eukaryotes were classified into other GH families and analysis indicate that some derive from an ancient ancestor the nematode cellulases consistently showed significant similarity (BLAST E-values < 10−10) to homologues from a range of bacteria, including plant-pathogens. Similarity searches and subsequent phylogenetic analysis resulted in seemingly credible trees in which nematode cellulases clustered as nearest neighbours of certain groups of bacterial cellulases. Hence, it was hypothesized that GHF5 genes have been acquired via HGT from bacteria. The only other eukaryotic cellulases next to the nematode genes in GHF5 originated from different yeast species.

Contrary, the pine wood nematode Bursaphelenchus xylophilus, a pathogenic species that is unique in its ability to feed on live trees and fungi, harbours GHF45 cellulases. GHF45 cellulases have been found in fungi, bacteria, protists, and a very small number of animals. The ancestor of B. xylophilus is likely a non-pathogenic Bursaphelenchus (which are solely fungal feeders) and the pathogenic B. xylophilus GHF45 phylogenetically is more closely related to the fungal cellulases than to ones found in insects. Hence, it was hypothesized that B. xylophilus cellulases are of fungal origin, and acquired by HGT. In both of the above-mentioned cases, GHF5 and GHF45 cellulases, BLAST-based similarity searches were the first indication that the gene may have been acquired by HGT from bacteria to nematode or from fungi to nematode. Furthermore, using similarity searches it has been suggested that GHF45 cellulases are absent from root knot and cyst nematodes, and similarly GHF5 cellulases have not been detected in B. xylophilus. While the GHF5 cellulases have been detected in five root knot and four cyst nematode species, the absence of sequence data from the other two completely unexplored suborders of the order Tylenchida and from other Bursaphelenchus species



precludes using the distribution pattern to strengthen the HGT claim. GH28-polygalacturonases of bacterial origin have only been found to date in RKN and the closely related Pratylenchidae and may represent a more recent HGT event. Polygalacturonases of family GH28 have also been reported in Aphelenchus avenae, but phylogenetic analysis indicates a distinct origin and a higher similarity to fungal GH28 proteins. Furthermore, it is commonly considered that plant parasitism does not reflect the lifestyle of the last common nematode ancestor. Therefore, it is possible that two independent HGT events from the same or closely related source bacteria underlie the presence of these genes.

Functions: Four functional roles have been described for the PPN genes thought to have been acquired by HGT:

1. Modification and degradation of plant or fungal cell walls: The plant cell wall consists mainly of cellulose fibrils cross linked by hemicelluloses and embedded in a matrix of complex polysaccharides called pectins. All identified PPN cell wall–modifying enzymes are capable of breaking down one or more of these cell-wall components. Cellulases act on beta-1,4 glycosidic bonds of cellulose, pectate lyases and polygalacturonases cleave alpha-1,4 linkages of pectate xylanases can degrade xylan (the major component of hemicelluloses), arabinogalactan galactosidases and arabinases are thought to hydrolyze beta-1,4-galactan in the hairy regions of pectin, and expansin-like proteins (found in X. americanum) weaken the non covalent interactions between cellulose and hemicellulose, thus exposing these components to the activity of degrading enzymes. All these proteins are expressed in the nematode’s subventral gland cells and allow the parasite to break the plant cell wall (in combination with the mechanical puncturing of the stylet) during invasion or soften the cell wall in order to allow migration intercellularly (as is the case for Meloidogyne spp.).

2. Suppression of host plant defences: Some PPN are biotrophic pathogens that feed on live

plant cells for several weeks and therefore need to suppress host defenses for the duration of their life cycles. It has been suggested that some genes acquired by HGT may be important in this process, either as detoxifying agents or as true suppressors of host defense signaling. Candidate cyanate lyases have been identified in the genomes of both M. hapla and M. Incognita, which helps in the suppression of host defense mechanism.

3. Establishment of the nematode feeding site: One of the horizontally acquired genes that might be involved in the establishment of the nematode’s feeding site is similar to NodL genes from rhizobia. Rhizobia are nitrogen-fixing symbiotic soil bacteria that form nodules on the roots of their leguminous hosts. Nodule formation is a complex process requiring lipo chitooligosaccharide signals from the bacteria (the Nod factors). NodL encodes an N-acetyltransferase involved in the biosynthetic pathway of Nod factors. RKN produce an as-yet-uncharacterized compound that induces changes in roots similar to those induced by Nod factors, and mutant leguminous plants that cannot support normal symbiosis with nodulating bacteria are less susceptible to infection by RKN, suggesting some overlap between the signaling pathways underlying symbiosis with nodulating bacteria and nematode feeding-site establishment.

4. Nutrient biosynthesis and processing: A total of nine different genes involved in the synthesis or salvage of vitamins B1, B5, B6, and B7 have been identified in CN, and two of these genes are also present in RKN. B vitamins are essential to all living organisms, but while absorbing these vitamins plants may restrict the availability of b vitamins as a defense mechanism. Therefore, horizontally acquired genes for biosynthesis and salvage of B vitamins may allow the nematode to circumvent this defense mechanism. Invertases, which are generally absent in animals, convert sucrose— the major form of sugar circulating in plant tissue—into glucose and fructose, which can readily be used as a carbon source by the nematode (found in RKN).

ENTOMOLOGY

19804

73. A Novel War Dance of the HoneybeeR E KARTHICK*, M. S. SAI REDDY, SOMALA KARTHIK AND MANOJ KUMAR

Department of Entomology, Dr. Rajendra Prasad Central Agricultural University, Pusa *Corresponding Author Email: [email protected]

Honey bees are the eusocial insects which plays a major role in pollination. They communicate among

themselves about location and distance of food availability through dance language. They dance



to signify their nest mates about the distance and direction of source of nectar, propolis and pollen which includes round dance and waggle dance. Recently, it was observed that the honey bee, Apis cerana japonica, a subspecies of Apis cerana, exhibit a different type of dance namely the war dance to warn the Asian giant hornet which preys on honey bees.

Asian Giant Hornet, Vespa Mandarinia japonicaIt is the natural enemy of Apis spp. and it preys the bee colony during autumn. The adult hornet preys on bees and crushes it into a paste with its powerful mandibles, to feed the young larva. The gaster is restricted only for the liquid food hence the adult is not capable of digesting the solid protein. The adult hornet obtains vespa amino acid mixture in exchange to the paste. The mixture produced by the larva is rich in amino acid. V. m. japonica is the only species among the social wasps that has the ability to apply pheromone to guide its nest mates regarding the food source. The sixth sternal gland secretes the pheromone i.e. Van der vecht’s gland. This behaviour is common during the autumn season where the hornet goes for scouting and hunting the honey bees.

Hive Entrance DanceApis cerana japonica initially performs waggle movement near the entrance and alerts its nest mates to collect the plant materials other than the flower resources and smears it on the entrance of its hive. This behavior occurs immediately after scouting by hornets. Hive entrance smearing can be interpreted as a means of masking the hornets’ forage-site marking pheromone using the strong odor of original or oxidized secondary materials collected from plants belonging to polygonaceae and lamiaceae, which are known to contain odorant substances that repel insects.

HypothesisTo test this hypothesis, Ayumi Fujiwara, Masami Sasaki and Izumi Washitani (2015) performed a series of attack simulation experiments with V. m. japonica

to check whether the war dance is a specific response to V. m. japonica scouting.

In addition to V. m. japonica, the suspected inducer of dancing behaviour, the responses to yellow hornet, Vespa simillima xanthoptera and yellow vented hornet, Vespa analis insularis, which occur sympatrically with A. c. japonica and are known to periodically attack honeybee hives were examined. In the attack simulation experiments, the number of dances at the hive entrance was counted. Then, both the dancing and dance-follower bees were examined whether they smear the hive entrance, and measured the abdomen-waggle times in each dance and the foraging-flight durations of the dancing bees.

To record the bee behavior, hives were placed in two apiaries namely apiary A and apiary B. The two hives in apiary B are subjected to experimental attack by V. m. japonica, and four hives in apiary A were subjected to experimental attack by V. s. xanthoptera and V. a. insularis. The apiaries were placed 4 km apart to exclude the influence of the V. m. japonica forage-site marking pheromone.

The dancing behaviour in honey bees was observed in case of V. m. japonica attack, whereas the dancing behavior was not noticed in case of V. s. xanthoptera and V. a. insularis attack. The actual foraging distance of A. c. japonica for collecting nectar and pollen ranges about 1-2 km. Whereas, the foraging distance of bees to collect odoriferous material in response to the dance ranges about 5-180m, which clearly depicts the quick action of bees in order to protect its colony.

ConclusionV. m. japonica scouting induces hive entrance dances in A. c. japonica. When one or a few workers perceives hornet scouts, she or they perform the entrance dance i.e. it waggles at the entrance. The war dance warns the nest mates of an “emergency” and alerts them to collect odoriferous plant materials used for smearing at the hive entrance from relatively nearby environment. It serves as a counter-attack strategy to deter V. m. japonica from attacking the colony.

20095

74. Weather based Pest ForecastingNEERU DUMRA*

Ph.D. Scholar, Dept. of Entomology, CCS HAU, Hisar- 125001 *Corresponding Author Email: [email protected]

Weather information is critical for developing pest models and decision tools that are useful for the success of an Integrated Pest Management program as it allows the grower to prepare for the ample possibilities of attacks that may occur. It requires that growers understand the kinds of pests that are common on the crops being cultivated throughout the growing season and learn about the life cycles of pests. Pest

forecasting guides the farmers regarding the timing of pest incidence to eliminate any possibility of blanket application of pesticide, efficient use of pesticides and reduce pesticide application rates (Mahal et al 2011). Pest forecasting models are highly effective in predictions of crop yield in advance of harvest and also forewarning of pests which integrates the statistical procedures like ANOVA, factorial analysis, regression



and multiple regression. For a pest management expert, prediction of pest population level along with its timing is important for planning and decision making (Maelzer and Zalucki 2000). Mahal et al (2011) describes about pre-requisites need to develop pest forecasting models which requires following basic information1. Quantitative seasonal studies which involves

seasonal abundance and sampling of pest population

2. Life history and pest biology which involves life span, survival rate, food, intrinsic growth rate in field as well as in-vitro conditions

3. Ecological studies of the pest which includes life table studies of the pest which is important for better understanding of pest population build up, natural mortality factors and critical growth stages

4. Crop Phenology which includes different crop cultivars, fertilizer dosages, irrigation, cultivation practices and plant spacing which influence the phenology of the crop

5. Natural enemies which involves the population of natural enemies present on various time intervals in the crop

6. Agro-ecosystem which involves changing cropping pattern and crop diversification involving different crops with a wide range of varieties of different maturity groups serve as suitable niche for supporting the buildup of Helicoverpa

Pest Forecasting Models

Forecasting model for mustard aphid in PunjabDhaliwal et al (2005) conducted study in Ludhiana from 1988-89 to 1997-98 to evaluate the use of agro meteorological indices for forecasting mustard aphid (Lipaphis erysime) on Raya, Brassica juncea. A correlation was provided for estimating the relationship between weekly aphid population and humid – thermal ration. There was high infestation in the year 1992-93 and 1996-97, with aphid populations ranging from 700 to 1300 aphids / plants. A pest weather diagram constructed during the high infestation years shows that lower minimum temperature, high morning relative humidity and reduced solar radiation or sunshine hours favoured the aphid population. The historical data on mustard aphid incidence and mean temperature were analyzed from 1988-89 to

2003-04. Mean temperature during January for both low and high aphid infestation years were compared. During the higher aphid infestation year, the mean temperature during January remained below 130C.

Forecasting Model for Helicoverpa armigeraThe monthly data of moth catches from pheromone trap during 1983-90 along with weather parameters of the corresponding period at PAU, Ludhiana was analyzed at National center for Integrated Pest Management, New Delhi by Das et al (2000) and it was found that there were two major peaks of moth catches in a year. The first peak was observed during March-April in chickpea crop and second peak during October of every year in cotton crop. The peak population of Helicoverpa armigera moth during March-April and October can be predicted in advance by multiple regression models using different weather parameters along with previous season’s population.

The population density during March-April (PM-A

) had been regressed with different weather parametres as well as pest population density of previous five months separately and cumulatively. P

M-A that depends

on total moth catches per trap during previous October to February (P

O-F), mean monthly Relative Humidity

recorded in the afternoon during previous February (RHE

F) and mean monthly minimum temperature of

previous February (TminF

) as follows

PM-A

= − 1032.65 + 2.06 PO-F

− 34.26 RHE

F + 516.74 T

min F (R2=0.75)

PM-A

= Population density in March and April

PO-F

= Moth catch per trap from previous year during October to February (P

O-F)

RHEF Monthly mean relative humidity

during previous February Tmin

F = Mean monthly minimum

temperature of previous February

Similarly, the population density during October (P

O) can be predicted in advance with multiple

regression equation using weather parameters as well as pest population of preceding months as follows:

PO

= − 5.30 +.12PM-A

+ 1.28 PJ – 0.51 R

J-J (R2=0.87)

PJ

= Pest population of June R

J-J = Total amount of rainfall during

June and July

TABLE: Various Pest forecasting models around the world

Forecasting Models Insect Parameters Country Reference

Ordinal logistic Model

Whitefly, Pyrilla and Fruit fly

Max temp., Min temp. and RH

India Agrawal and Mehta (2007)

CLIMAX Helicoverpa sp. Temperature and humidity Australia Zalucki and Furlong (2005)SOPRA Dysaphis plantaginea

and Grapholitha lobarzeweskii

Air and soil temperature Switzerland Graf et al (2002)



Forecasting Models Insect Parameters Country Reference

FLYPAST Aphis fabae Suction trap data UK Knight et al (1992)NAPPFAST Scirtothrips dorsalis Degree days and cold

temp. survivalUSA Nietschke ‎(2008)

ReferencesAgrawal R and Mehta SC (2007) Weather Based

Forecasting of Crop Yields, Pests and Diseases - IASRI Models Journal of Indian Society of Agricultural Statistics 61: 255-263

Das DK, Trivedi TP, Singh J and Dandapani A (2000) Weather based prediction model of Helicoverpa armigera for Integrated Pest Management in cotton-chickpea based agro-ecosystem of Punjab. Paper presented in International Conference on Managing Natural resources for Sustainable Agricultural Production in 21st Century. Voluntary Papers Natural Resources, Vol 2. Feb14-18,2000, New Delhi, pp 621-622

Dhaliwal LK, Hundal SS and Kular JS (2005) Use of agrometeorological indices for forecasting of Mustard aphid (Lipaphis erysimi). Journal of Agrometerology 7: 304-306

Graf HU, Hopli PH, Hohn PH and Blaise (2002) SOPRA: a forecasting tool for insect pests in apple orchards. Acta Hortculture 584: VI International Symposium on Computer Modelling in Fruit Research in fruit Research and Orchard Management

Knight JD, Tatchell GM, Norton G and Harrington R (1992) FLYPAST: an information management system for Rothamsted Aohid Database to aid pest control research and advice. Crop Protection 11: 419-26

Maelzer DA and Zalucki MP (2000) Long range forecasts of Helicoverpa in using the SOI and the SST. Bulletin of Entomological Research 90: 133-146

Mahal MS, Singh B, Sarao PS and Singh S (2011) Role of insect-pest forecasting in integrated pest management In: Arora R, Singh B and Dhawan A K (eds), Theory and Practice of Integrate Pest Management, Scientific Publishers, Jodhpur, India, pp 248-261

Nietschke BS, Borchert DM, Magarey RD and Matthew AC (2008) Climatological potential for Scirtothrips dorsalis (Thysanoptera: Thripidae) establishment in the United States. Florida Entomology 91: 79-86

Zalucki MP and Furlong MJ (2005) Forecasting Helicoverpa populations in Australia: a comparison of regression-based models and a bioclimatic based modeling approach. Insect Science 12: 45-56

20106

75. Ecological Threats for Pollinators and their ConservationTARA YADAV AND RICHA BANSHIWAL

Ph.D. Scholar, Division of Entomology, RARI (S.K.N.A.U.), Durgapura *Corresponding Author Email: [email protected]

Human population are increasing day by days, demand for food also increasing with it. To cope with this, in the future, our agricultural systems will need to produce more food in a sustainable way. Pollinators are vital factor to these systems. Pollinators are external agents which help in the transfer of pollen grains from one flower to another of the same or another plant of the same species. Relations between insect- pollinators are very important for the sustainability of agricultural ecosystems. It has been estimated that over 80 per cent of all flowering plants depend on insect pollinators, especially bees (Raj, 2017).

Threats for PollinatorsMany agricultural practices impact directly or indirectly pollinator populations. Human activities and farm practices are major factors in the loss of habitats of pollinators leading to a decrease in their food supplies (pollen and nectar). Several features associated with modern agriculture make farms poor habitat for wild

bees and other pollinators. Pollinator crisis seems more in landscapes dominated by annual crops. Some weeds provide forage for pollinators. Removal of these weeds is a key factor in the decline of native pollinators in agro ecosystems (Richards, 2001). Climate change will alter the close relationship between insect pollinators and the plants that depend upon them for reproduction. Non-native species can compete with native plants or animals for resources. Air pollution causes threats for bees and other pollinators that rely on scent trails to find flowers. Pesticide misuse and drift from aerial spraying are a major threat to insect pollinators, especially spraying persistent chemicals. These chemicals can kill bees and other pollinators directly or cause a variety of sub lethal effects such as impairing their ability to find their hive or provide food for their larvae. Systemic insecticides applied to seeds can contaminate the pollen grains that are an essential source of food for bees and their young. Light



pollution can harm moth pollinators by increasing their susceptibility to predation by bats or birds when they are attracted to artificial lights at night.

Pollinator’s Conservation1. Habitat manipulation: Understanding the

biology of flowers and the behavior of pollinators, it is important to understand how to manage agro ecosystems in order to provide nesting habitat as well as continuing alternative sources of forage that can sustain populations of pollinator’s year around.

2. Flowers: Planting a succession of crops that flower at different times could greatly enhance pollinator abundance. Create a continuous flow of nectar and pollen from early spring through late fall. Encourage combinations of annuals and perennial plants. Growing variety of flower colors and shapes to attract pollinators.

3. IPM: Adoption of integrated pest management for increasing of pollinators. Minimizing pesticide use along with implementing other agricultural practices; protects water resources from pesticide runoff, minimizes exposure of people, pets and

wildlife to pesticides and provides stable long-term pest control instead of the frequent boom and bust pest cycles associated with preventive use of broad-spectrum pesticides. Do not spray pollinator-attractive plants with insecticides when open flowers are present.

4. Optimize of herbicides: Herbicides will kill important native plants such as milkweed that pollinators rely upon as a food source and a place to raise young. Make the commitment to avoid using chemicals and to maintain your garden in a natural, organic way.

5. Regular supply of water: Due to scarcity of water flowers are wilting and less availability of food for pollinators.

ReferenceRaj, H. (2017). Various threatening factors to the

biodiversity of insect pollinators in Himachal Himalaya, India. International Journal of Sciences & Applied Research, 4(7): 22-35.

Richards AJ (2001). Does low biodiversity resulting from modern agriculture practice affect crop pollination and yield? Annals of Botany, 88: 165–172

20133

76. Biology and Management of Cigarette Beetle, Lasioderma serricorne (F.)JAI HIND SHARMA AND GAURAVA KUMAR

Department of Entomology, College of Agriculture, G. B. Pant University of Agriculture and Technology, Pantnagar-263145 (Uttarakhand)

IntroductionThe cigarette beetle, Lasioderma serricorne (F.) (Coleoptera: Anobiidae), is a pest of considerable economic importance in several countries of the world. The species is the most destructive of the insect pests of unprocessed and processed tobacco and extremely large variety of other materials of both plant and animal origin.

General ClassificationKingdom: Animalia, Phylum: Arthropoda, Class: Insecta, Subclass: Pterygota, Order: Coleoptera, Suborder: Polyphaga, Family: Anobiidae, Subfamily: Xyletininae, Genus: Lasioderma, Species: Lasioderma serricorne.

Life Cycle

EggOpaque pearl white coloured; oval or oblong shaped eggs, varying from 0.29 to 0.50 mm long and 0.18– 0.25 mm in diameter are laid directly on the dried food material within 12 h after mating. Eggs turn yellowish shortly before hatching. The total number of eggs laid

by a single female in a lifetime varied ranging from 10 eggs to 100 eggs or more, having an oviposition and post oviposition period of about 7 d each.

LarvaDuring hatching, the larva chews its way through the chorion (outer shell of an insect’s egg) at the posterior margin of the papillated (protuberance) region, so that it lifts like a cap enabling the larva to emerge through the opening. The newly emerge larva may eat the whole egg shell and other unhatched eggs if no other food is available. Firstly, newly hatched L. serricorne larva is semitransparent, but progressively becomes whitish or yellowish in color. On emergence, the larva (active for 4-5 d without feeding move in search of food) is very active and is capable of crawling considerable distances in search of food. Thus, infestation may easily spread from infested to uninfested materials. The young larvae are negatively phototrophic, that is, would back away from light. They will enter small holes in search of food and can penetrate packaged commodities. L. serricorne consist of four instars before pupation. During the second molt, the larvae assume the characteristic scarabaeiform (grub shaped



and their bodies are curled to form a shape like the letter C) shape. When fully grown, larvae become turgid, immobile, and stops feeding. It then forms a thin cell or cocoon made of food waste material cemented together by secretion produced by the midgut. It is found four larval instars at 30°C and 70% RH, while the number of molts increased from four instars between 22.5 and 30.0°C to five instars at 20°C. Larvae are quiescent and stop feeding about 24 h before each molt.

Prepupae and pupaeThe matured larvae go quiescent in the cocoon before pupation, where it undergoes structural changes preparatory to pupation known as “prepupal”. At 30°C, the matured larva cast its skin after 2–4 d in the cocoon. The prepupal stage is not distinguishable from the matured larva by any other distinct external characteristics. Before transforming to pupa, the matured larva or prepupa undergoes a slight contraction in length losing it scarabaeiform shape to become perfectly straightened, with the body becoming more deeply wrinkled.

Pupae are uniformly white with a slight greenish tinge when they are newly formed, but gradually assume a reddish-brown color, which darkens with age. Sexual dimorphism is present at the tip of the abdomen, which can be seen after the molted skin is removed. The terminal segment of the female pupa has a pair of lateral lobes, which are very distinct and are absent in the male. The pupal stage lasts 7 d in the summer, but 10–14 d in cooler weather.

AdultThe adult stays in the pupal cells from 4 to 6 d before sexually matured, and with fully developed coloration. Both sexes of the adult are similar in external appearance. In general, the body is elongate-oval, moderately convex, pubescence moderate, and sub-recumbent. There are 11 segments on the antennae. Courtship and mating is aided by both pheromonal and tactile stimuli provided by the female, which are perceived by respective sensilla of the antennae and palpi of the male. Female shows polyandry (mating with several males).

DamageIn tobacco leaves, larvae tunnel out long cylindrical galleries especially near the midrib, while in ground food, eggs are laid on the surface, and resulting larvae feed down into the flour in all directions. Adults of the beetle cut holes to penetrate or escape from packaged commodities leaving a neat round hole. Larval feeding causes direct damage to foodstuffs and non-food items. Larvae will sometimes bore their way through cardboard boxes and other packaging in search of a place to pupate. In the tobacco industry, losses due to L serricorne infestations are estimated to be about 0.7–1% of the total warehoused tobacco commodity. The important thing is that their damage by consuming commodity is very less and the actual loss is by: 1) weight losses; 2) reduced market value of

manufactured products (cast-off-skins, dead bodies, gnawed particles, and other products); and 3) loss of goodwill.

ManagementSanitation is the first barrier in the control of L. serricorne, maintaining cleanliness in the factory, wholesale, or retail establishment, including destruction waste material, damaged stock, etc., in which the beetles may breed.1. L. serricorne, are good fliers, and can easily infest

by moving in and out of facilities, installation of screens (openings less than 1.0 mm) on doors, windows, and other openings in the warehouses and manufacturing facilities to check insect movement into and out buildings and to increase the effectiveness of other control measures. Screens can also be incorporated with contact insecticides to provide extra advantage.

2. Insecticide-treated nets not only act as a physical barrier, to keep away the unwanted insect pests, they can also act chemically by repelling or killing the insects that come in contact with the net. Commodities should be disinfested carefully (e.g., fumigation, extreme temperatures), before placed under the nets.

3. Monitoring of insects pests is helpful in pest management decision making, for that one can use conventional monitoring devises for stored-product insect pests include light traps, physical devices (e.g., electric grid, fly tapes), and food or chemical attractants. Pheromone (serricornin) traps placed 1.5–2.0 m above the floor is proved helpful in its monitoring.

4. Low temperatures: (shifting the storage facility to regions of the country/state where the daily mean temperature is 5°C for over 70 consecutive days in a year) can reduce the infestation to considerable level.

5. Heat treatment: under normal atmospheric pressure treatment of tobacco leaves with steam at about 8.7 bars to raise the temperature to 115.6°C, which quickly kills all stages of the cigarette beetle. In case of established infestations of L. serricorne within facilities, control can be achieved through heat sterilization. Heating commodities to 54–60°C for 18–30 h (40 h generally) will kill all life stages of the target pest.

6. Phosphine fumigation will destroy all insect life stages in most types of packaging, while it enables a residue-free treatment. (long term Phosphine use can have corrosive effect on copper containing materials ex. electrical wiring in buildings and electronic circuitry)

7. (S)-methoprene having 33.6% a.i. can be used for crack and crevice treatments in warehouses and processing facilities. (an analogue to juvenile hormone, prevents the development of the pupa into the adult)

8. A contact insecticide (e.g., pyrethrins or pyrethroids) in the form of aerosols or fogs of will kill the beetles that are hit by the spray particles



(Sodium ion channels modulator). This type of treatment is only effective against insects when is hit by its droplets. Spraying with the contact insecticides should be done in dusk. (in day, flight activity of the insects is at peak)

9. At 24 to 26°C and 90–95% RH, DE (Diatomaceous Earth) at 500 ppm causes 100% mortality of L. serricorne populations within 5 d exposure period.

10. Irradiation of L. serricorne, at 100 Gy to 300 Gy can reduce its population in processed products.

11. Controlled atmospheres: ≥20% CO2 and/

or ≤1% O2, with the rest of the atmosphere composed of N2 gas. This technology has not been widely adopted by the tobacco industry because of capital and running costs of the equipment.

12. Mating disruption: synthetic sex pheromone (serricornin) is released into available space which makes it difficult for males to find females for mating, by this means causing population suppression or extinction. Product consists of a dispenser that holds ample quantities of the sex pheromone.

20141

77. Beeswax, a Miracle of Bee Hive: Its Production, Compositions and UsesSASWATI PREMKUMARI*, SABUJ GANGULY AND RASHMI MANOHAR MAHALLE

Research Scholar, Department of Entomology and Agricultural Zoology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi- 221005, UP, India *Corresponding Author Email: [email protected]

IntroductionHoney bees are the beneficial insects which help in pollination of most of the plants. Without bees, pollination would be difficult and time consuming - it is estimated that one-third of the human food supply depends on insect pollination. These are social insects that live in colonies. The hive population consists of a single queen, a few hundred drones, and thousands of worker bees.

Honeybees produce a number of substances like honey, pollen, propolis, royal jelly, and beeswax, all of which are used by human being for nutrition and medicinal purposes. Here we will discuss more about the beeswax, how it is produced by the bees and how it is useful for bees itself as well as for the human beings.

What is Beeswax and how it is ProducedIt is a wax produced by honey bees of the genus Apis. Worker bees that are younger than 18 days old are the best wax producers. It is secreted from eight wax-producing glands present in the inner sides of the sternites of 4th and 7th abdominal segments of worker bee. After 18 days, the glands start declining until the death of the worker bees. The wax appears as transparent small flakes on the bees’ abdomen. The bee then uses the stiff hairs on their hind legs to remove the scales of wax from its abdomen and then pass on it to the middle leg and then take to its mouth where it starts to chew on it by the mandible. After being chewed, it become white. In this mastication process the salivary secretions are added to the wax to make it softer. During this process, the beeswax picks up small portion of honey, pollen, and propolis which darkens its color. During its peak wax production

phase, a healthy worker bee can produce about eight scales of wax in a 12 hour periods.

Composition of Bee WaxThe chemical composition of the bees wax is a bit complicated. It contains about 15 different chemical compounds, which are divided into three groups: free fatty acids (13.5 to 15%), esters (70.4 to 74.7%) and saturated hydrocarbons (12.5 to 15.5 %). It also contains at least 284 different compounds, mainly a variety of long-chain alkanes, acids, esters, polyesters, and hydroxy esters etc. It is also rich in vitamin A (4096 IU/ 100 g). It is generally resistant to hydrolysis and natural oxidation and is insoluble in water. It is solid at room temperature and becomes brittle at temperature below 180C. It has a specific gravity of about 0.95 and a melting point of over 140 degrees F.

Uses of Bee WaxBeing a natural wax, it is a valuable commodity that beekeepers can harvest and sell for many commercial uses. It has hundreds of uses among which few are listed below.

� Bee wax is mainly produced by the bees for building their combs in which they rear their brood and store honey and pollen for their future need.

� In cosmetics: around 40 per cent of the world trade in bee wax is used for the cosmetic industry. These are used for making moisturizer, lip balm, hair creams, hair conditioner etc. When used in skincare products, it hydrates, soothes, repairs, and fortifies the skin. Also used for making beeswax-infused deodorant bar with a probiotic effect.



� In pharmaceutical preparation: around 30 per cent of the world trade in bee wax is used for the pharmaceutical industry. These are used for the preparation of drugs, pills, capsules etc and is also used as the main ingredient in the preparation ointments and creams used to treat burns and wounds and to soothe joint pain. Beeswax plays an important role in Ayurvedic medicine, the ancient and traditional Indian medicine, with the name of “Madhuchishtha” (Patwardhan, 2014).

� In candle making: around 20 percent of the beeswax trade is used for candle making. Beeswax candles exhibit air-purifying properties. Unlike paraffin candles, they help decrease the number of airborne contaminants, such as bacteria, allergens and dust. But these candles are less common and more expensive than paraffin candles.

� In medicinal application: In apitherapy, inhalation of beeswax is used to treat upper respiratory disorders. Having regenerative quality and anti-inflammatory property, it helps to decrease the irritation, redness, and inflammation characteristic of acne, while its anti-septic effect further facilitates the healing process. It also used to treat eczema and rosacea. It prevents harmful bacteria from entering the body through chapped

and broken skin and it provides the skin with a layer of protection against external irritants (Fratini et al., 2014)

� Other uses: used to make polish for cars, furniture, shoes and for treatment of other leather products. Used as lubricants for industrial use. Used for making drawing crayons. Also used for making figures and sculptures.

ConclusionBee wax is a natural wax produced by the honey bees. Although these are intended for the use of the bees themselves, human beings have successfully used these products for their own benefit in a wide spectrum of applications. This is a clear-cut proof that in nature we can find all that we need for our life, health and treatment of illness.

ReferencesFratini, F., Cilia, G., Turchi, B. and Felicioli, A. (2016).

Beeswax: A minireview of its antimicrobial activity and its application in medicine. Asian Pacific Journal of Tropical Medicine, 9(9), 839-843.

Patwardhan, B. (2014). Bridging Ayurveda with evidence-based scientific approaches in medicine. EPMA Journal, 5(1), 19.

20182

78. Lesser Grain Borer, Rhyzopertha Dominica: A Major Threat to Storage Grains in IndiaR. TAMILSELVAN1, BANKA KANDA KISHORE REDDY2, J. KOUSIKA3 AND S. KARTHIKEYAN4

Ph.D. Scholars1,2,4 and Post-doctoral Fellow3

Department of Entomology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. *Corresponding Author Email: [email protected]

Based on Type of Grains Damaged: Primary Storage Pest

Based on position of damage- Internal feeder

Origin � R. dominica thought to be originated from Indian

subcontinent and first observed from Australia in 1896, but now it has cosmopolitan and polyphagous distribution (Edde, 2012).

� It is also found in temperate countries due to its ability for prolonged flight or as a result of international trade in food products.

Host RangeMajor hosts

� Adults and larvae of R. dominica feed primarily on stored cereal seed including wheat, maize, rice oats, barley, sorghum and millets

Minor hosts � Beans, dried chillies, turmeric, coriander, ginger,

cassava chips, biscuits.

Biology and ecology � Females of R. dominica lay between 200 and 500

eggs in their lifetime laid on grains, gunny bags, or cracks and crevices.

� The lowest temperature at which R. dominica can complete development is 20 o C (Egg to adult-90days).

� The faster rate of development occurs at 34 o C and 5-6 generations



(1) Egg-2 days (2) Larval -20-55 days (3) Puape-3 days (4) Adult –live for 40 to 80 days

(Srilakshmi and Virani, 2018)

Similarities with other species

Character Lesser grain borerR. dominica

Large grain borerP. truncatus

Shape of abdomen

Square with distinct corners

Tapered abdomen

Elytra Tapered and rounded

Slopes towards the tip is steep and flat

Colour Brown Dark brownLength 2-3mm 3-4 mm

Damage Symptoms � It is a primary pest. � Adults and grubs feed on wheat, rice, maize,

sorghum, groundnut, dry fruits. � Feed on internal content and grain converted into

shells with irregular holes. � Adults produce frass, thus spoiling more than they

eat and convert grains into flour � The maximum damage by this pest occurs during

July to October

Management � A study conducted by Jhumar et al., 2015 revealed

that grains treated by diatomaceous earth @ 1.0% was found to be most effective with significantly highest adult mortality (99.5%), least grain damage (0.25%), least weight loss (0.13%) and least population build up (0.75 adults).

Biological Control-ParasitoidsMost of the parasitoids that attack the rhizopertha

were belongs to the families PteromalidaePteromalids - solitary larval ectoparasitoidsChoetospila elegansAnisopteromalus calandraeDinarmus basalisLariophagus distinguendusPteromalus cerealellaeTheocolax elegans

BethylidaeCephalonomia rhizopethae (Binoy, 2015)

Entomopathogens � The pathogenicity of entomophaghous fungi

depends upon various physical (temperature, relative humidity, application time of fungal insecticide, dark and light period etc.) and biological factors (the specific host species, host pathogen interaction etc.).

� Beauveria bassiana (Ascomycota: Hyphomycetes) � Metarhizium anisopliae (Ascomycota: Sordario) � Purpureocillium lilacinum � Isolates of Purpureocillium lilacinum was the best

in controlling target insect species (Barra et al., 2013).

Chemical ControlCentral Insecticide Board recommendationsFumigants Dose PeriodAluminum Phosphide 56% 150 g/100m3 5 daysAluminum Phosphide 15% 900g/100m3 5 daysAluminium Phosphide 77.5% GR

3.35g/m3 7 days

Methyl Bromide 98% W/W 24g/m3 5 daysEthylene Dichloride + Carbon Tetrachloride 3:1

300- 400gm/m3

(230-307 ml)48-72 hrs

� Deltamethrin 2.5% WP (for grains and seeds in stacks) @ 30 mg a.i /sq.m

ReferencesEdde P. A. 2012. A review of the biology and control of

Rhyzopertha dominica (F.) the lesser grain borer. Journal of Stored Products Research, 48: 1-18.

Srilakshmi, C and Virani V.R. 2018. Biology and Behaviour of Lesser Grain Borer, Rhizopertha dominica (Fabricious) (Coleoptera: Bostrichidae) on Stored Wheat in Laboratory Conditions. International Journal of Agriculture Sciences, Vol:10 (5):5231-5234.

Jhumar, L., Sharma, K. C and O.P. Ameta. 2015. Efficacy of inert dusts against lesser grain borer, Rhizopertha dominica Fab. In stored wheat. Annals of Plant Protection, 23 (2): 231-236.

Binoy, C.F. 2015. Hymenopteran parasitoids associated with some important stored product pests in Kerala, India. International Journal of Science, Environment and Technology, Vol. 4 (2): 509 -513.

Barra, P., Rosso, L., Nesci A and M. Etcheverry. 2013. Isolation and identification of entomopathogenic fungi and their evaluation against Tribolium confusum, Sitophilus zeamais, and Rhyzopertha



dominica in stored maize. Journal of Pest Science, 86(2):217-226.

20188

79. EffectsofPesticidesonWildlifeBANKA KANDA KISHORE REDDY1, N. S. A. DEVI2, J. KOUSIKA3, S. KARTHIKEYAN4 AND R. TAMILSELVAN5

Ph.D. Scholars1,2,4,5 and Post-doctoral Fellow3

Department of Entomology and Dept of Agril. Microbiology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. *Corresponding Author Email: [email protected]

IntroductionPesticide residues in wildlife as an indication of the potential hazard to wildlife from pesticides. Greater in fish eating birds such as gulls or bald eagles and use of DDT caused egg shell thinning in many birds. Exposure to highly toxic pesticides cause sickness or death. Once sick wildlife may neglect their young, abandon their nests and become more susceptible to predation and disease.

Indirect EffectsAdverse effects caused by modification or elimination of wildlife habitat or food supply Herbicides can reduce food, cover and nesting sites for wild life Insecticide can reduce insects- that serve as food supply for other animals. Plant pollination can be affected by reduction in population of bees and other pollinators.

Aquatic EnvironmentFish accumulate residues gradually (much slower than mollusks), but their pesticide levels change when fat is mobilized. Oysters and other shellfish not only accumulate chlorinated hydrocarbon pesticides rapidly but also discharge them when they are returned to pesticide-free media (Butler, 1967a,b). More than 150 estuarine stations have been established to monitor pesticide levels on the coasts of the United States by using indicator organisms. The choice of earthworms as an indicator is also excellent in view of their importance in the food chain for bird populations. However, the relationship between the amount of pesticide intake and the amount appearing in certain tissues or products is better understood in aquatic animals. Water that contains particles high in organic matter tends to make lipophilic pesticides less available to fish.

Bottom mud of high organic content readily. binds pesticides, in contrast to sandy or rocky bottom materials. It is not uncommon that fish from clean lakes contain more DDT than ones from lakes with higher degrees of eutrophication (Cope, 1966). Macek and McAllister (1970) found that among 12 species of fish representing five different families, salmonids were always the most susceptible to all insecticides tested (nine chlorinated hydrocarbon, organophosphate, and carbamate insecticides); ictalurids and cyprinids

were the least susceptible. Fish are known to have very inefficient mixed-function oxidase systems to detoxify these insecticides, which makes them vulnerable to them as environmental contaminants.

BirdsEggshell thickness of wild birds generally decreased as the levels of environmental contamination increased. Thinner eggshells, which result in egg breakage, have been suggested as the reason for the declining reproduction rate in several bird species showed a general decline. Porter and Wiemeyer (1969) examined American sparrow hawks kept on diets containing 0.28 and 0.84 ppm of dieldrin and 1.4 and 4.7 ppm of DDT. These birds produced eggshells roughly 10% thinner than normal ones. Other commonly observed patterns among raptorial populations, such as breakage and disappearance of eggs, were also noted.

Later, Wiemeyer and Porter (1970) fed a diet containing 2.8 ppm of p,p’-DDE and in the second year again observed a 10 % reduction in the thickness of eggshells. While there are other conflicting reports, it appears safe to state that some of the chlorinated hydrocarbon insecticides must cause a reduction in eggshell thickness in some bird species. There appear to be susceptible species and resistant species as far as eggshell thinning is concerned. There are many reports that claim inhibition of carbonic anhydrase, generally acknowledged to play an important role in forming eggshells, by DDT and some other derivatives. The carbonic anhydrase activity in the shell gland itself can be affected by DDT and DOE, as judged by an in vitro assay in experiments with birds fed a diet containing DDT or DOE. Degrees of inhibition found were 60 %. O,p’ -DDT has been reported to show estrogenic effects at high doses, and both o,p’-DDT and p,p’-DDT have anti-estrogenic effects in the rat (Clement and Okey, 1972).

1. Carbonic anhydrase in the shell gland of avian species is unusually sensitive to chlorinated hydrocarbon insecticides

2. In vivo inhibition observed is a result of some other effect, such as reduced enzyme production or the production of substances that suppress the activity of the enzyme.



Acute PoisoningShort exposure to some pesticides may kill or sicken wildlife. Fish kills caused by pesticides residues carried into waterways by run off, drift. Bird kills- pesticide treated vegetation, insects, pesticide granules and bait or treated seed. Direct parameter for measuring pesticide effects on wildlife is acute LD50 value. Fish are generally very susceptible to pyrethroids and chlorinated hydrocarbon insecticides. Mammals are more sensitive to OP and carbamates (Cope, 1971). Diflubenzuron are toxic to aquatic invertebrates such as copepods, Diaptomus spp., cladocerans & amphipods (Ali and Mulla, 1978).

Chronic PoisoningExposure of non-lethal levels of pesticides over extended periods can cause reproductive effects. Population of bald eagles and other birds of prey were reduced by widespread use OC (DDT) in 1950’S and 1960’s. These compounds and metabolites caused reproductive effects in birds. Reduction in use of OC in 1970’s and early 1980’s resulted in greatly improved reproduction & increasing bird population. The long-term toxicity (chronic) of an insecticide may be quite different from its short-term toxicity.

Indicate the ratio between acute LD50 and chronic minimum lethal dosage. A low ratio can be regarded as the sign of a noncumulative poison (e.g., 2.4 for Zectran ®), since it means that the daily dose (about one-third of the total dose) is not much less than the amount of pesticide needed to kill the animal in one exposure. In such a case, the animal is effectively eliminating the poison through detoxification or excretion mechanisms at the end of each day. In contrast, high ratios such as for DDT, dieldrin, and endrin indicate the cumulativeness of chlorinated hydrocarbon insecticides.

TABLE: Ratio between acute oral LD50 and chronic minimum lethal dosage of various insecticides for mallards

Chronic minimum lethal doges

(EMLD) (mg/kg/day)

Ratio of cumulativeness (Acute LD50/

EMLD)DDT 50 44.8Dieldrin 1.25 76b

Endrin 0.125 45Abate® 2.5 32-40Dursban® 2.5 30Parathion 3-6c 2.7-5.3c

Sevin® (Carbaryl) 125 17.4Baygon® (arprocarb)

2.0 2.0

Zectran® 1.25 2.4

Factors Influencing ToxicityAge and SizeAge and size apparently are the two most important factors influencing susceptibility. We generally assume

that susceptibility to pesticides decreases as the animal grows, and much of the evidence supports this concept. This is very much evident in fish, for which insecticide toxicity has been assessed by means of the concentration changes in the ambient water (not the amount given per unit body weight, as with other animals). For example, the water in Lake Michigan contains around I ppt of DDT. but even this low concentration is toxic enough to affect the hatchery operation for coho salmon. Bigger fish are not affected by it, so they have been imported into Lake Michigan for planting coho salmon colonies.

Laboratory examinations of DDT toxicity to various stages and sizes of coho salmon (Buhler and Shanks, 1970) have confirmed that older fish and, within the same age group, bigger fish are more resistant to DDT (i.e., the median survival time is directly related to body weight). There are many other experimental data indicating the susceptibility of smaller fish. Older fish in turn accumulate more pesticides than the younger ones. Young birds are quite vulnerable in the path of a pesticide spray, or spray drift. Without a protective feather cover the young bird is doubly jeopardized.

Among fish species on which considerable bioassay data are available, larger fish are less susceptible than smaller ones in the 0.5 to s-g range. Some pesticides are highly toxic to fish eggs, while other are not. For example, antimycin, a piscicide, killed rainbow trout eggs at 19 ppb. Body size and toxicity from ingested DDT has been investigated for coho salmon, and two factors were seen to be important. Laboratory reared salmon of the same age were fed a diet containing DDT, and the median survival time was found to be directly related to body weight, perhaps because the smaller fish lack sufficient lipid to handle adequate storage detoxification of the DDT (Buhler and Shanks, 1970).

SexThe sex of the animal often makes a difference in susceptibility to pesticides. The acute oral toxicity of aldrin, chlordane, DDT, and heptachlor are higher for male white rats than for females, while with endrin, endosulfan, mirex, toxaphene, and some others the females are more susceptible. Among organophosphates, acute oral toxicity is higher to male white rats for Abate®, Azodrin®, methyl parathion, and some’ others, but higher to females for carbophenothion, coumaphos, Diazinon®, EPN, malathion, and parathion. Among carbamates, carbaryl has higher oral toxicity to females; for others, sex seems to make no difference. With dermal application both organophosphates and chlorinated hydrocarbons are more toxic to females than to males.

Pesticide FormulationFor fish, insecticide emulsions and oil solutions are the most toxic kinds of formulations. Rainbow trout, red salmon, and three-spined stickleback in Alaska succumbed faster to emulsified and oil solutions of DDT, BHC, toxaphene, and chlordane than to acetone suspensions. Emulsifiable oil preparations of benzene hexachloride were 25 times more toxic to



golden shiners than wettable powder formulations containing the same level of gamma isomer. Granular formulations, wettable powders, and dusts, which usually release the active ingredients into the aquatic ecosystem at slow rates, have relatively low toxicities to fish. Thus, choosing formulations of these kinds can often afford protection to animals in the aquatic environment.

TemperatureEntomologists have long known of negative temperature coefficients with some insects and DDT and methoxychlor, with increased mortalities at lower temperatures. This is also seen in some fish; methoxychlor shows negative temperature coefficients with rainbow and bluegills.

Chemistry of the EnvironmentBottom muds of high organic content bind pesticides through adsorption, in contrast to sandy or rocky bottoms. Such adsorption often binds significant proportions of the amount entering the water, thus lowering the concentration in the water and reducing the hazard to fish. While the pesticide is attached to the substrate, it may degrade, may be slowly released to the water, or may persist unchanged for years, depending upon the chemical, the nature of the substrate, and the chemistry of the water.

It has been suggested that the concentration of toxicants in ponds can be reduced by the addition of silt or some other substrate to the water to adsorb

the chemical and make it less available to fish. The toxicity of pesticides may depend upon water chemistry. DDT in waters of high pH is not very toxic to salmonids. Malathion hydrolyzes rapidly in alkaline waters, but slowly in neutral or slightly acid waters. Another important consideration is the species-specific differences in the rate of insecticide pickup. For instance, Lumbricus terrestris accumulates each insecticide at a level slightly higher or lower than that found in soil, while Allolobophora chlorotica consistently accumulates much higher levels of all the insecticides.

ReferencesAaron, T.F., Paul, F.H., Jean-Marc, G., Jason, D., Ross,

J.N., Keith, A.H., Michael, K., Derek, C.G.M., 2003. Influence of habitat, trophic ecology and lipids on, and spatial trends of organochlorine contaminants in Arctic marine invertebrates. Mar. Ecol. Prog. Ser., 262: 201–214.

Bevenue, A., Hylin. J. W., Kawano, Y., and Kelly, T. W. (1972). Organochlorine pesticide residues in water, sediment, algae and fish, Hawaii 1970-71. Pesric. Monir. J., 6: 56.

Coble, Y., P. Hildebrandt, J. Davis, F. Raasch, and A. Curley (1967). J. Am. Med. Assoc.202: 153

Fumio Matsumura.1985. Toxicology of Insecticides. Plenum Press, New York. P. No:444-479.

Weisburger, J. H., and G. M. Williams (1980). In Toxicology, 2nd ed. J. Doull, C. D. Klassen, and M. O. Amdur, eds. Macmillan, New York, p. 84.

20192

80. Method Validation for Pesticide Residues AnalysisS. KARTHIKEYAN1, BANKA KANDA KISHORE REDDY2, J. KOUSIKA3 AND R. TAMILSELVAN4

Ph.D. Scholars1,2,4 and Post-doctoral Fellow3

Department of Entomology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. *Corresponding Author Email: [email protected]

Evaluating a method to ensure that its performance is appropriate for the analysis being carried out. In general, validation should be checked that the method accomplishes adequately for the purpose throughout the range of analyte concentrations and test materials to which it is applied.

Procedure for Establishing a Method to be ValidUniversal requirements to test its performance for a method follows as

� Linearity � Accuracy � Precision � Range � Recovery � Limit of Detection � Limit of Quantification

� Ruggedness or Robustness

LinearityIt verifies that the response is linearly proportional to the analyte conc. in the conc. range of the solution and Performed using std solution at five conc. level in the range of 50 -150 % of the target analyte concentration each std should measure at least three times or more.

Linearity data are often judge from the coefficient of determination (r2) and the y intercept of the linear regression line. An r2 value > 0.998 is considered as evidence of acceptable fit of the data to the regression line.

AccuracyIt is the closeness of the obtained value to the true



value of the sample. It can obtain through recovery studies. The acceptable recovery should be in the range of 80-120%.

PrecisionPrecision of analytical method is obtained from multiple analysis of a homogenous sample and Precision data are obtained by one laboratory on one day, aliquots of the homogeneous sample that have been independently prepared.

RangeIt is the concentration range over which acceptable accuracy and precision are obtained. The validated range is the interval of analyte concentration within which the method can be regarded as validated.

RecoveryRecovery is the amount measured as a percentage of analyte (active substance and relevant metabolites) originally added to a sample of the appropriate matrix, which contains either no detectable level of the analyte

or a known detectable level.Recovery percentage=Area of sample x Vol. made upto (mL) x Standard

conc.(ng) x 100Area of standard Vol. injected (μl) Wt of the

sample (g) Spiked level

Limit of DetectionIt is the lowest amount of an analyte in a sample that can be detected but not necessarily quantified as an exact value.

Limit of QuantificationThis is the lowest of analyte that can be measured in the sample matrix at an acceptable level of precision and accuracy. An acceptable precision 10-20% relative standard deviation depending on the concentration levels measured.

Ruggedness or RobustnessIt will identify those factors that will contribute to variability of the results and should not be changed.

20201

81. The Sterile Insect TechniqueHARPREET SEKHON* AND RAVINDER NATH

Department of Entomology, School of Agriculture, Lovely Professional University, Jalandhar, Punjab *Corresponding Author Email: [email protected]

The sterile insect technique (SIT) is an eco-friendly insect pest control method successfully used throughout the world. SIT involving the mass-rearing of target pest and then sterilizing them by using radiation (i.e. gamma rays and X rays and chemosterilants), after that sterilized male population is systematically released over a large area through air or manually over specified areas, where they compete with wild males of the same species in terms of foraging, flight and mainly mating, After mating with the wild females, non-viable zygote formation leads to non all hatching of eggs and target pest population starts declining and gets eradicated in due course from the target area with the cooperation of the local residents/farmers. Density-dependent effects will reduce, eliminate, or even reverse the beneficial effect of a SIT program. SIT does not involve any transgenic (genetic engineering) processes.

E. F. Knipling was the very first person to conceive an approach to insect pest control using Sterile Insect Technique (SIT) by successfully controlling screwworm fly (Domestic cattle pest). Knipling reared and released sterile insects into the environment in large numbers (10 to 100 x). After that native female mated with a sterile male and produced fertilized but sterile eggs and in the 5th generation, he got complete control over the pest population. After the success of screwworm fly eradication using SIT program many other agencies worldwide were led to the investigations on this

radio genetic technique for control of many other economically important pests. International Atomic Energy Agency (IAEA) runs Coordinated Research programs & Technical projects to promote SIT and encourages scientists globally to be a part of it.

SIT has been successful only after when it is integrated with other Integrated Pest Management (IPM) practices, for the management of major economically important insect pests, counting various fruit flies species viz. Mediterranean fruit fly, Mexican fruit fly, oriental fruit fly, melon fly, Tsetse fly, screwworm, many lepidopterans viz. codling moth, pink bollworm, false codling moth, cactus moth, and the Australian painted apple moth and mosquitoes. Latest SIT program (2017-18) on Aedes aegypti is in progress in Queensland, Australia where they released >3 million males sterilized with the natural bacteria Wolbachia and results showed 80% reduction of the population in trial area.

SIT technique has been first developed in United States of America and has been used successfully for over 60 years and it has been currently applied in 6 continents and on reviewing its economic assessment, it has demonstrated very high economic returns on globally. Not only this, SIT being an environment friendly technique doesn’t produce any kind of environmental pollution and with this it also helped in reducing pesticide consumptions. In addition to this



crops and livestock production losses have also come down. SIT technique have safe guarded horticultural and livestock industries from Insect pest losses, eventually resulted in increased productivity and market price.

Main Components of Employing ‘Sterility Principle’ (I.E. SIT)1. Mass rearing: Rearing of target pest (Males) in

large numbers in the laboratory.2. Treatment: Doses of treatment must be

standardized. Low doses are always preferable.3. Competitiveness: Sterile male should be able to

compete with wild males in every aspect specially in mating.

4. Release: How many insects are to be released at the time of release and how many releases are enough for adequate control of pest is very important.

5. Evaluation: After release impact of the experiment must be assessed.

6. Re-infestation: At the end of season some population of wild males will remain in field so, to avoid their population buildup few sterile males should be released at the end of season.

AdvantageEnvironmentally friendly: SIT will control the target insect pest species with negligible off-target effects on other insect species. The only non-target effects are indirect, for example, the potential effect on other members of the ecosystem of removing or suppressing one species. During this program no lethal chemicals or chemical residues are released or deposited in the environment.

DisadvantageOne species at a time: Granting species specificity SIT is immensely prominent from an environmental perception, it may have downsides where several pest species need to be controlled at once. SIT is thus best suitable in the areas with a single dominant insect pest species needs to be controlled, more than one would be controllable, but more than two would point to a more broad-spectrum approach.

Sterile insect technique (SIT) is a component of Integrated Pest Management not a solo method. It cannot eradicate pest all alone. So, before starting any SIT program cultural, physical, mechanical, Biological and chemical methods should be applied and when pest population is 50% down then this method can naturally eradicate the pest from the target area.

The International Plant Protection Convention has classified sterilized insects as beneficial insects. SIT program differs from the classical biological control programs, which includes the augmentation of non-native biological control agents, in quite a few ways:

� Sterile insects cannot multiply on their own and hence they cannot establish themselves in the environment.

� It is also called autocidal control, because it

breaks the target pest’s reproductive cycle and by definition it is species-specific.

� The SIT program does not introduce non-native species into a new ecosystem.In case of India, keeping in view about the

increasing demands of eco-friendly control measures, it is expected that Sterile insect technique (SIT) will progressively gain much more importance in upcoming years and following is the list of ongoing SIT programmes in India:

� Spodoptera litura (Tobacco caterpillar) (Delhi Univesity),

� Culex pipiens fatigans (WHO, MRC, DRDO), � Rhyncophorus ferrugineus (Red palm weevil)

(BARC), � Maruca vitrata (Redgram Webber) (UAS,

Raichur).

Feasibility of SIT for LepidopteraSIT is limited in case of Lepidoptera because they are highly radio resistant. They have holokinetic chromosomes which makes them highly radio-resistance. So, due to this they require large doses for 100% sterility. Associated with somatic damage and behavioral incompetence in P-1 (parent generation) lead to limited success by SIT

In Diptera, fully sterile gamma dose is given to the male, which mates with normal female, produce inviable zygote giving complete sterility. Whereas, in case of Lepidopterans F-1 sterility technique is used successfully i.e. giving very low gamma doses inducing partial sterility in males. When this male mate normal female then two cases happen (1) Inviable zygote (exhibiting dominant lethality), (2) Viable Zygotes (carrier of recessive lethality). Then F-1 generation mates with normal female then it produces inviable zygote giving complete sterility.

ReferencesMaru, N. K., & Sao, Y. (2018). Sterile Insect Technology

for Pest Control in Agriculture, International Journal of Bio-Resource and Stress Management, 9(2), 299–305. https://doi.org/10.23910/ijbsm/2018.9.2.3c0896

Alphey, L., Benedict, M., Belini, R., Clark, G.G., Dame, D.A., Service, M.W., Dobson, S.L. (2010). Sterile-Insect Methods for Control of Mosquito-Borne Diseases: An Analysis, Vector Borne and Zoonotic Diseases, 10(3), 295-311. doi: 10.1089/vbz.2009.0014



20207

82. The Story of Monolake and Alkali FlySAFEENA MAJEED, A. A.

Ph. D. Scholar, University of Agricultural and Horticultural Sciences, Shivamogga *Corresponding Author Email: [email protected]

California’s Mono Lake, the shallow lake which is about 13 miles outside ‘Yosemite National Park’, has been considered as the “most inhospitable lake”. The lake is three times saltier than the ocean, often with a pH of 10 or above. In addition, it is filled with laundry detergents like borax and sodium carbonate, the combination of which makes Mono Lake slippery and hence considered as a death sentence for many organisms. But it is not true for the alkali fly (Ephydra hians, Diptera: Ephydridae), which loves breeding and feeding in this difficult environment. This genus is known to inhabit extreme environments like alkaline/saline lakes, acidic thermal springs, coastal marshes and tidal splash pools (Herbst, 1990), but particular species adapted to specific habitats, suggesting physiological specialization for living in unusual chemical environments (Herbst et al., 1988).

The larva has an unusual physiological adaptation to combat the accumulation of carbonate and bicarbonate ions in the blood. Modified malpighian tubules unlike other insects, the lime glad (fig. 1) in alkali fly larvae partially removes carbonate ions from the blood. Within these specialized lime gland tubules, excess carbonates are precipitated with calcium, forming calcium carbonate and stored in the lime gland till moulting, in order to fulfill the dietary need for calcium, as Mono Lake has extremely low calcium concentrations (Herbst et al., 1989).

It is more difficult for other flies to escape from Mono Lake water, due to the high concentration of Na

2CO

3 which penetrates and wet their cuticle. This

effect creates a small negative charge at air-water interface, which generates an electric double layer facilitating wetting. But, upon entering the lake, alkali flies are protected by an air bubble formed around their super-hydrophobic cuticle. This trait arises from a combination of factors like a denser layer of setae on cuticle and prevalence of straight-chain alkanes like C24, C27 (fig. 2) in the cuticle (Breugel et al., 2017).

FIG. 1: III instar larva of E. hians, arrows indicating lime gland (Scale: 1 mm)

FIG. 2: GCMS analysis of hexane extracted cuticular hydrocarbons of alkali fly.

Ephydra hians larvae are osmoregulators (Herbst et al., 1988) capable of maintaining haemolymph osmolality in media with an osmotic concentration 10 times higher than that of the blood capable of both hyper- and hypo-osmotic regulation of haemolymph osmolality. But this ability was less effective in seawater/sodium chloride suggesting that larvae are alkali-adapted and that restriction in habitat distribution not more than salinities above 200 g/L has a physiological basis.



Table 1. Lethal salinity tolerance concentrations and exposure times for Mono Lake E. hians larvae

LC50

TIME (h)TDS

MONO LAKE WATER SEA WATER/ NaCl

mOsm TDS mOsm48 273 6550 183 5780

72 252 6030 158 4970

96 190 4520 144 4550

LT50

MONO LAKE WATER SEA WATER/ NaCl

Salinity LT50 Salinity LT50

g/l mOsm hours g/l mOsm hours

200 4760 88.5 - - -

250 5980 76.0 193 6110 43.5

300 7200 40.5 - - -

None of these brine flies is endemic; they inhabit distinctive habitats of different salinity levels, chemical compositions etc. forming the basis for biogeographic distribution patterns (Herbst, 1999). Within any habitat, changing salinity conditions over time known

to impose physiological or ecological constraints due to their specific habitat preference and adaptations and further alter the population progression patterns and can create imbalance in the relative abundance of co-inhabiting species. The latter must be taken care seriously because of the so-called changing land use pattern, urbanization etc. after all these tiny flies are an important component in the process of eating and being eaten (benthic algae → alkali fly → migrating birds).

ReferencesBreugel, F.B. and Dickinson, M.H. (2017)

Superhydrophobic diving flies (Ephydra hians) and the hypersaline waters of Mono Lake. PNAS. 114 (51):13483-13488.

Herbst, D.B. (1990) Biogeography and physiological adaptations of the brine fly genus Ephydra (Diptera: Ephydridae) in saline waters of the Great Basin. Great Basin Naturalist 59(2):127-135.

Herbst, D.B. and Bradley, T.J. (1989) A malpighian tubule lime gland in an insect inhabiting alkaline salt lakes. J. Exp. Biol. 145:63-78.

Herbst, D.B, Conte, F.P. and Brookes, V.J. (1988), Osmoregulation in an alkaline Salt Lake insect, Ephydra (Hydropus) hians Say (Diptera: Ephydridae) in relation to water chemistry. J. lnsect. Physiol. 34 (10):903-909.

INTEGRATED INSECT PEST MANAGEMENT

20089

83. Endophytes in Plants DefenceBHAGYASREE S N, SURESH M NEBAPURE AND SAGAR D

Division of Entomology, ICAR-Indian Agricultural Research Institute, New Delhi-110012 *Corresponding Author Email: [email protected]

Sustainable era of agriculture is anticipating organic alternatives to chemical insecticides for plant protection, as the use of chemicals is upsetting the ecosystem balance by polluting soil, air and water, causing pest resurgence, secondary pest outbreaks and resistance besides affecting the economy of the country by hindering export because of residue. To overcome this, our focus has shifted to utilize ecosystem services provided by bioagents, especially fungi and bacteria as they are known to affect wide array of chewing and sucking pests. But their commercial application is limited due to their susceptibility to abiotic factors viz., temperature, humidity and ultraviolet (UV) rays. To overcome these bottlenecks, a hidden ecological role played by bacteria and fungi as endophytes was sought. As the endophytes symbiotically live inside the plants thus gets protection against abiotic stresses and UV rays furthermore play a defensive role against insect pests and diseases. Association of endophytes and plants has a mutual benefit which includes:

Benefits Endophytes gets from Plant1. Supply of nutrients2. Protection from environmental stress3. Passive transfer and spread between host

Benefits Plants gets from Endophytes1. Drought resistance2. Improved growth response3. Insect resistance4. Anti-herbivore to mammals5. Production of compounds which are of having

biotechnological interest

Why Endophytes are Asymptomatic??Endophytes are different from plant pathogen for the nutrition, they are not competitive and toxic, usually pathogen compete with the host cell for their nutrition results in diseases, but endophytic microorganisms do not compete, they utilize host exudates, dead cortical cells, excess carbohydrates, plant debris for their



nutrition during the stage of infection and during the time of colonisation they use components of symplast and apoplast movement.

Scope of EndophytesDespite of 4 decades of research with thousands of publications proving their ecological significance, they are not clearly characterised, virtually endophytes are known to produce plethora of the substances which includes antibiotics, antimycotics, immune suppressants and anticancer compounds, for example: Taxol: anti-cancer drug; Cryptocin: antifungal agent; Jesterone: antifungal; Oocydin: antifungal; Isopestacin: antioxidant; Munumbicin: wide spectrum antibiotic and Kakadumycin: antibiotic, which can be potential exploited in various field like,1. Pest management2. Crop production3. Disease management4. Livestock production and management5. Medicine and pharmaceutics industry6. Other allied fields

Benefits of Artificial Colonisation of EndophytesIn case of pest management, numerous reports have shown that fungi and bacteria have the capacity to colonise artificially through various methods of inoculation and control insect pests (Azevedo et al., 2000). In past two decades, a great deal of information is also available on the artificial endophytic association of Beauveria bassiana (Balsamo) Vuillemin in various commercial crops which includes, maize (Bing and Lewis, 1991), tomato (Ownley et al., 2004), opium poppy (Quesada-Moraga et al., 2006), date palm (Go´mez-Vidal et al., 2006), bananas (Akello et al., 2007), coffee (Posada et al., 2007) and common bean (Parsa et al., 2013). However, it is becoming apparent that relationships between plants and their endophytic micro-flora are more complex and the endophytes can be involved in the production of growth promotion factors and mediators that induce the plant’s natural resistance (Rodriguez et al., 2009) and they are advantageous because1. Effective against cryptic feeders like stem borers

and gall making insects.2. Resistant to biotic and abiotic stress3. Little inoculum is required4. No need of frequent application5. Development of virulence is less6. Development of mutation is nil

Identification of EndophytesThere are four methods presently in use for detecting and identifying fungi and bacteria in plant tissue1. Histological observation most recently in

combination with molecular methods.2. Surface sterilisation of the host tissue and

isolation of the emerging fungi on appropriate growth media.

3. Detection by specific chemistry, e.g. immunological methods.

4. By direct amplification of fungal DNA from colonised.

Mode of DefenceSecondary metabolite production: Some major classes of secondary metabolites are found to be produced by endophytic fungal associations with plants which include indolediterpenes, ergot alkaloids, peramine and related compounds. The terpenes and alkaloids are also produced by endophytic plants which show inducible defenses and act similarly to defensive compounds produced by plants and are highly toxic to a wide variety of insect pests.

Altered nutrient content: Endophytes are known to alter the C:N ratio of leaves and making them a less efficient source of proteins and affect the chewing insects. Additionally, endophytic association increases the availability of limiting nutrients to plants improves overall performance and health, potentially increasing the ability of infected plants to defend themself against all kinds of biotic stress.

Production of enzymes: endophytes are known to alter the enzymes such as superoxide dismutases (SOD), catalases (CatA), peroxidases (POD), alkyl hydroperoxide reductases (AhpC), and glutathione S-transferases (GSTs) to enhance the plant defence against insect pests.

Despite of various kinds of research carried out on endophytes, they have been underutilized due to bottlenecks limiting the use of endophytes in agriculture as well as poor recognition of endophytic mode-of-action, so in order to make them commercially viable, research should focus on narrowing these gaps.

ReferencesAkello, J., Dubois, T., Gold, C.S., Coyne, D.,

Nakavuma, J. and Paparu, P., 2007. Beauveria bassiana (Balsamo) Vuillemin as an endophyte in tissue culture banana (Musa spp.). Journal of Invertebrate Pathology, 96(1):34-42.

Azevedo, J.L., Maccheroni Jr, W., Pereira, J.O. and de Araújo, W.L., 2000. Endophytic microorganisms: a review on insect control and recent advances on tropical plants. Electronic Journal of Biotechnology, 3(1):15-16.

Bing, L.A. and Lewis, L.C., 1991. Suppression of Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae) by endophytic Beauveria bassiana (Balsamo) Vuillemin. Environmental Entomology, 20(4):1207-1211.

Gómez-Vidal, S., Lopez-Llorca, L.V., Jansson, H.B. and Salinas, J., 2006. Endophytic colonization of date palm (Phoenix dactylifera L.) leaves by entomopathogenic fungi. Micron, 37(7):624-632.

Ownley, B.H., Pereira, R.M., Klingeman, W.E., Quigley, N.B. and Leckie, B.M., 2004. Beauveria bassiana, a dual-purpose biocontrol organism, with activity against insect pests and plant pathogens. Emerging Concepts in Plant Health Management. Research Signpost, India, pp. 255-269.

Parsa, S., Ortiz, V. and Vega, F.E., 2013. Establishing fungal entomopathogens as endophytes: towards endophytic biological control. Journal of Visualized



Experiments, JoVE, (74).Posada, F., Aime, M.C., Peterson, S.W., Rehner, S.A.

and Vega, F.E., 2007. Inoculation of coffee plants with the fungal entomopathogenBeauveriabassiana (Ascomycota: Hypocreales). Mycological research, 111(6):748-757.

Quesada-Moraga, E., Landa, B.B., Muñoz-Ledesma, J.,

Jiménez-Diáz, R.M. and Santiago-Alvarez, C., 2006. Endophytic colonisation of opium poppy, Papaver somniferum, by an entomopathogenic Beauveria bassiana strain. Mycopathologia, 161(5):323-329.

Rodriguez, R.J.; White, J.F., Jr.; Arnold, A.E.; Redman, R.S. 2009. Fungal endophytes: Diversity and functional roles. New Phytol.,182:314–330.

POST-HARVESTMANAGEMENT

20087

84. EffectofProcessingTechnologiesonFingerMilletSUSRITA SAHU AND RUKEIYA BEGUM

Scientist (Home Science) and Scientist (Plant Science), KVK Bargarh, OUAT

INTRODUCTION: Finger millet (Eleusine coracana) belongs to the family Poaceae. Globally, 12% of the total millet area is under finger millet cultivation, covering more than 25 countries of Asia and Africa with limited economic resources. Being an agronomically sustainable crop, it can grow on marginal lands, high altitudes and can easily withstand drought and saline conditions, requires little irrigation and other inputs and yet maintain optimum yields. In India, it is one of the important millets occupies highest area of cultivation among the small millets and more commonly known as ragi or madua.

Nutritional Composition: This is considered one of the most nutritious cereals which is easy to digest. It contains about 81.5% carbohydrates, 5–8% protein,15–20%,1-1.7%fat,18-20% dietary fiber, and 2.5 -3.5% minerals. Of all the cereals and millets, finger millet has the highest amount of calcium (344 mg%) Apart from this it also contains potassium (408mg), Phosphorous (283 mg), Iron (3.9 mg), Vitamin B

1 (1.71mg), Vitamin E (22 mg). 100 grams of Finger millet has an average of 336 KCal of energy in them. It also contains phytochemicals such as tannins (0.04-3.47%), phytate (0.48%), oxalate (0.27%), cyanide (0.17%), saponins (0.36%) and phenolics (0.3–3%) (Rathore et al, 2019).

Due to its high dietary nutritional & phytochemical content, it can provide numerous health as well as and therapeutical benefits such as Anti-diabetogenic, anti-inflammatory, anti-tumerogenic anti-aging, antiulcer, atherosclerogenic and antimicrobial properties. However, the anti-nutritional effects of phytochemicals are partly negative because they reduce the digestibility of nutrients and the absorption of minerals which restricts the application of finger millet in food preparations. Moreover, to get better shelf life of finger millet flour along with processed foodstuffs reduction of anti-nutritional features is essential. Hence it is necessary to study the effect of different processing technologies to bring the anti-nutritional factors into the permissible limits for utilization of finger millet in a better way.

Soaking: It is commonly used technique in order to clean & wash foreign particles. Soaking of ragi in water (1:10) for 1-2 days at room temperature (25˚C) reduces polyphenols, phytate, saponins, oxalates and trypsin inhibitors which ultimately improve bioavailability and bio accessibility of minerals and also the nutrition quality.

It has also reported that ragi soaked in NaOH solution or distilled H

2O for 8 hours significantly

reduces tannin content. It also reduces phytic acid content ranges from 39.47 to 24.17% which increases the bioavailability of minerals like zinc.

Cooking: Cooking upon water or steam also inactivate heat labile anti-nutrients in ragi. The hydrothermal treatment of finger millet improved the carbohydrate digestibility of finger millet increased from 61 to 73 g/100 g and protein digestibility from 79 to 91 g/100 g (Dharmaraj and Malleshi,2011).

Fermentation: Fermentation has great impact on enhancement of net protein utilization (NPU), niacin, thiamin, riboflavin and biological value (BV) of finger millet. In fermented finger millet, Lactobacillus salivaricus increases lysine and tryptophan content by 7.1% and 17.8%, respectively. Use of endogenous grain micro flora in finger millet flour during fermentation shows a significant reduction in the amount of antinutrient factors such as trypsin inhibitor activity by 32%, tannins by 52% and phytates by 20%.

Germination: It leads to expansion of α and β-amylase, which develops desirable aroma during roasting makes it a supreme grain for malt foods. It also helps to reduce the anti-nutritional content of finger millet as tannin, phytic acid and trypsin inhibitors activity which drastically improve bio-accessibility of dietary minerals such as zinc, iron and calcium. It helps in increasing in lysine & methionine content from 3.5 to 4.0 mg/100 and 1.3 to 1.5 mg/100 g respectively in germinated finger millet. The tannin content of finger millet was reduced to 20%, 45%, 62% and 72% after germination of 0, 24, 48, 72 h respectively.

Malting: It is a combined process of steeping, germination, drying, toasting, grinding and sieving



in order to achieve high nutritional quality, better starch digestibility, sensory properties and reduced antinutritional activities. The malting leads to reduction of 41 to 33% phytate in finger millet. Malting of finger millet carried out at 48 h/30 0C increase the protein content from 6.42 g/100 g to 7.32 g/100 g

Decortication: The decortication of finger millet is difficult due to presence of seed coat attached to fragile endosperm. Therefore, decortication of finger millet is done through hydrothermal treatment such as hydration, steaming and drying which increases hardness of endosperm texture and nutritional contents of finger millet grains. Thus, decortications have a significant effect on reduction of phytate phosphorus and polyphenols contents by 39.8% and 74.7%, respectively which ultimately enables the nutritional bioavailability of protein and minerals.

Roasting: During roasting, the anti-nutritional or toxic effect such as saponins, alkaloids, glycosides, gioterogenic agents, tryptin inhibitor and hemagglutinin are removed. Processed foods obtained as a result of roasting of finger millet grains helps in increases the bioavailability of iron

Puffing or popping: It is a traditional method used for producing ready-to-eat and stable shelf-life products which are crunchy and porous. It also involves soaking whole unhusked grains in water and mixing with sand heated at 250 °C for 15-60 s. Puffing on its part, increases the digestibility and solubility of starch due to gelatinization. Popping reduces certain anti-nutritional factors and most of the toxin effects for example hemagglutinin, cyanogenic glycosides, gioterogenic, saponins, alkaloids and trypsin inhibitors.

Extrusion: Extrusion cooking is work with the

principle of combined efforts of shear force along with high pressure and temperature which is responsible to modify starch properties. During the extrusion process, protein solubility and structure are decreased and disrupted, thus increasing in-vitro protein digestibility. Fortification of extruded products with minerals and vitamins is also employed to balance the nutritional composition that is lost during processing.

Radiation: Food irradiation technology is a process in which packed foods are subjected to controlled ionizing radiation in the form of x-rays, alpha, beta and gamma rays. It helps in preservation of foods by extending their shelf-life, improves the nutritional quality of foods and reduces anti-nutritional compounds.

CONCLUSION: The traditional processing technologies such as germination, soaking, fermentation, puffing and cooking reduce the level of phytochemicals with increasing the bioavailability of micronutrients. However, the new processing technologies and preparation methods are needed to enhance the bioavailability of micronutrients of this underutilized crop with better quality and varieties of commercial value-added products.

ReferencesDharmaraj U, Malleshi NG. Changes in carbohydrates,

proteins and lipids of finger millet after hydrothermal processing. LWT-Food Science and Technology. 2011; 44:1636–42.

Rathore T, Singh R, Kamble DB, Upadhyay A, Thangalakshmi S. Review on finger millet: Processing and value addition, The Pharma Innovation Journal 2019; 8(4): 283-291.

ECONOMICS

20121

85. Land Holding in India: An InsightSHUBHI PATEL1 AND ANJU YADAV2*

1Department of Agricultural Economics, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh 2Department of Agricultural Economics and Management, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur, Rajasthan Corresponding author- Anju Yadav

India is a land of agriculture dominated employment sector. Due to its rich geographical diversity, variety of crops are grown in India ranging from cereals, horticultural products, pulses, oilseeds, spices, and plantation crops etc. The returns from each crop depend on the type of crop grown like whether it is a cash crop which fetch high price or cereal which is sold at minimum support price. Also the returns are a function of the area cultivated. It is established that there is dominance of small and marginal farmers in India (Figure 1).

The figure 1 shows that, 86% of the farmers belong to the small and marginal land class. This means having land holding less than 2 Ha. This emerges as an issue because small land holding leads to less farm produce thus, weak bargaining power of the farmers. Its increases the cost of production, and transportation cost and less marketable surplus. While large farmers are only 1% out of the total farming population.

An insight into the growth of the average operational holding of different land classes shows that the average operational land holding for farmers



has reduced to 1.08 Ha in 2015-16 from 2.28 Ha since 1970-71 (Table 1).

Detailed class wise insight shows that the average land holding has reduced for all the land classes in India. At present the average land holding of Marginal farmer is 0.38 Ha, small farmer has 1.41 Ha, Medium farmer has 2.7 Ha and Large farmer has 17.1 Ha of land on an average.

The area operated also is of importance for the knowledge of which section of the land class holds how much portion of the total land operated (Figure 2). In this regard, highest percentage of land is held by marginal farmer i.e. 24% while large farmers operate only 9% of the total land operated. Since there is dominance of small and marginal farmers it is obvious that major portion of land is cultivated by this class i.e. 47%.

FIGURE 1: Number of operational holding: ALL INDIA, source Agriculture Census 2015-16

TABLE 1: Average size of Operational Holding All Groups: All India(in Ha)

Size group 1970-71 1976-77 1980-81 1985-86 1990-91 1995-96 2000-01 2005-06 2010-11 2015-16Marginal 0.4 0.39 0.39 0.39 0.39 0.4 0.4 0.38 0.39 0.38Small 1.44 1.42 1.44 1.43 1.43 1.42 1.42 1.38 1.42 1.41Semi medium 2.81 2.78 2.78 2.77 2.77 2.73 2.72 2.68 2.71 2.7Medium 6.08 6.04 6.02 5.96 5.96 5.84 5.81 5.74 5.76 5.72Large 18.1 17.57 17.41 17.21 17.33 17.2 17.12 17.08 17.38 17.1All India 2.28 2 1.84 1.69 1.55 1.41 1.33 1.23 1.15 1.08

Source- Agriculture Census 2005-06 and 2015-16

FIGURE 2: Area Operated: All India, source Agriculture Census 2015-16.

Thus, we can see that there is need to promote aggregation of farmers through FPOs, Co-operatives,

Kisan clubs in order to reap maximum benefit from the power of consolidation.

ReferencesAgriculture Census 2015-16. All India Report on

Number and Area of Operational Holdings Agriculture Census Division Department of Agriculture, Co-Operation & Farmers Welfare Ministry Of Agriculture & Farmers Welfare Government of India, 2019.



20202

86. Impact of Corona on Farm ProduceSHWETHAKUMARI U1* AND KIRAN S C2

1Ph.D. Scholar Department of Soil Science and Agricultural Chemistry, COA, UAS, Raichur. 2Ph.D Scholar Department of Forestry and Environmental Science, UAS, GKVK, Bengaluru. *Corresponding Author Email: [email protected]

The disruption in the supply chain of agriculture and horticulture produce, uncertainty over the next cultivation season, fall in demand and reduced consumption of food pose a threat to food security and nutrition owing to COVID-19 pandemic. Food and Agriculture Organisation had cautioned about this possibility if the governments did not take care of the food and nutritional needs of the marginalised. Especially the perishable foods like fruits, vegetables and flowers has gone unimaginable losses to farmers and even they are not ready to harvest the produce to avoid the expenditure which is more than the transportation of that produce to urban areas or to needy one. The states like in Karnataka farmers are incorporating the produce to soil by cultivating the land which has crops which is in harvesting stage keeping in mind that the produce will become a good manure to future crop. The produce price which is reaching to consumers is very high and there is no option to consumers, but when we see in deep the farmers are not getting at least 50 % of that amount, here comes the middle men. The best way to help farmers in this pandemic situation is by buying the produce and distribute it among the needy by the state government or it can connect farmers and consumers and notify traders to provide logistics support. Even those who are distributing food packets can include fruits and vegetables in kits. Form the consumer point of view the farmers are getting the good price but the actual situation is very different. The quantity of produce in market is less hence the price has gone up. The problems of farmers are numerous and in between this the AMPHANA is taking out all the

productive produce and the severity of amphana when it reaches landfall. The lockdown is severely disrupted India’s agricultural sector, now within extension being on the cord. There are worries that India’s most vulnerable will suffer. Without any government relief to restart farm activates. The agriculture sector employees nearly half of the workforce in the country, contributing 17.5% GDP while the government assured restarting of harvest many farmers who has standing Rabi crop fear uncertainty. In other hand absence or shortage of labourers or farm workers are facing entire family to salvage the produce. If the produce is not harvested in time then it will be destroyed or farmers won’t have anything to eat. In many parts of country farmers are staring huge financial losses, owing to untimely rain and hailstorms apart from lockdown blues and food supply chain messed up. Farmers are bearing a brunt of the nationwide lockdown with most crop left in out to rotted die especially onion farmers in Maharashtra and grape farmers in Karnataka are the worst hit. Huge heaps of grapes were discarded in Karnataka, Chikkaballpura district because there were no costumers. The central government says it has exempted transportation of harvest from the lockdown but the reality on the ground is that there are no workers and where there are workers there is no transportation. On the other side dairy sector is facing huge losses i.e., hundreds of litters of milk going down the drain. Here plays postharvest a major role and the only option to farmers to save the produce and to get good price.

COMPUTER ADDED TECHNOLOGY

20186

87. Mobile Apps in Agriculture1DR. M. KALPANA AND 2DR. R. PARIMALARANGAN1 Assistant Professor (Computer Science), 2Assistant Professor (Agrl. Economics) Dept. of Social Sciences, Anbil Dharmaligam Agricultural College and Research Institute, Tiruchirappalli, Tamil Nadu

IntroductionAgriculture is an important part in the Indian economy and human kind depends on agriculture as the major part of their livelihood. The farmers and their farm

activities play a vital role in the production of farm produces. The information is to be transferred to the farmers in the right time and in a cost-effective manner for their farming activities. Information and Communication Technologies (ICT) is a right way to



exchange the information to the farmers. There is an increase in use of mobile phones, one among the ICT tool for agriculture process. The mobile phone is used in all the areas of services and application.

Mobile app is used in the many areas of agriculture and allied sector for monitoring crop and its related activities. Mobile apps can be downloaded freely from online stores. There are many app developed for agriculture. In this article few apps are listed in detailed

PlantixPlantix is a mobile app developed to diagnosis the plant disease and is used for monitoring. In plantix app the pictures of affected plants are captured through the mobile phones and the app identifies the disease. This app is a real time monitoring tool for the pest and disease identification. The pictures are tagged with the real time monitoring of pest and diseases.

Iffco KisanIFFCO Kisan is the app which includes modules such as agriculture advisory, market price, weather and agriculture information in the form of images, text, audio and videos in language selected. The developed app supports eleven languages in India including English. The app also gives helpline number to have a touch with Kisan Call Services.

EnameNAM is an electronic National Agriculture Market, a trade portal which is promoted by the Government of India. This app is developed to create an integrated national market throughout India for agricultural commodities. eNAM helps to access the prices by the traders remotely through their mobile phones

AgrimarketAgriMarket app is developed to get the market price of crops within 50 km from the device location. The location of the mobile is fetched with GPS and gives the market price of crops around 50 km. Another way to get the price without the GPS location, the price are sourced from AGMARKNET portal. The AgriMarket is available both in Hindi and Tamil.

Digital MandiDigital Mandi is the app used to get the price of different agricultural commodities of different states

and districts in India. We can get the commodity wise or statewise details of agricultural commodity prices. We can also browse the commodity categories and select the specific commodity prices of various states.

RicexpertriceXpert is developed in English and Odia, an android based application. This app gives the latest technologies to the rice farmers for real time application in the field. This app gives the real time diagnosis of pest, diseases, weeds, nutrient deficiency and nematodes to farmers. Through this app the fertilizer recommendation is given to the rice farmers. The rice farmers can use as the diagnostic tool for rice crops, can query through test, pictures or record voice which would be addresses by the panel of experts for solution through SMS.

Kisan SuvidhaKisan Suvidha was developed by Government of India, Ministry of Agriculture and Farmers Welfare and provides information in Tamil, Hindi, Gujarati, Odia, Marathi and English. This app provides the information related to market price, weather details, plant protection, IPM practices, seeds, soil health card, expert advisory and cold storage.

Crop InsuranceCrop Insurance app provides the details of crop insurance. The mobile app helps to calculate the insurance premium of specified crops based on the area, loan amount and coverage amount. The app helps to get the details about the premium details, normal sum insured, extended sum insures and the subsidy information of specified crops.. The app is developed in English and Hindi

CONCLUSION: The mobile app is developed for the benefit of farmers and stakeholders. The output of the mobile app may either be text or videos. Now a day’s m-agriculture is the flourishing are in the field of agriculture. The mobile service helps to solve the communication gap between the farmers and extension workers. Through the mobile app the following information can be access for agriculture, such as price information, weather, pest and disease details, and nutrient management for the farmers for right time.



NATURAL RESOURCE MANAGEMENT

20179

88. Bioremediation Techniques for use of Sewage WaterVEERESH HATTI1, ASHOK K. SAINI2, SANJAY, M. T.3, NARAYAN HEBBAL4

*1Assistant Professor (Agronomy), Directorate of Research 2Assistant Professor (Agronomy), Centre for Natural Resource Management, SDAU, Sardarkrushinagar – 385 506 (Gujarat), 3Agronomist and Scheme Head, AICRP on IFS, UAS, Hebbal, Bengaluru – 560 024, 4Research Associate, ARS, Bidar *Corresponding Author Email: [email protected]

Rapid population growth and increased urbanisation over the years have led to quantum increase in generation of domestic sewage in India. The amount of sewage generated in most cities and towns of the country has exceeded the capacity of the available treatment systems and its disposal and treatment has become a challenge for the municipalities. In India, an estimated 38,354 million litres per day (MLD) sewage is generated in major cities, but the sewage treatment capacity is only of 11,786 MLD (Kaur et al., 2013). It is projected that by 2050, about 48.2 BCM (132 billion litres per day) of wastewater (with a potential to meet 4.5% of the total irrigation water demand) would be generated (Bhardwaj, 2005). It implies that there is a huge potential to treat and reuse the domestic sewage water for various purposes.

Sewage is a water-carried waste, in solution or suspension, also known as wastewater, is more than 99% water and is characterized by physical, chemical and biological constituents. Sewage comes from human waste, washing water, rainfall collected on roofs, yards, liquids from domestic sources (drinks, cooking oil, cleaning liquids) etc. The sewage water from the domestic areas joins natural water resources leading to water pollution, eutrophication, death of aquatic life, groundwater pollution etc.

Bioremediation:-The term contains two words i. e., “Bio” means “living” and “Remediate” means “to solve a problem”. The term implies the use of living organisms to solve an environmental problem. According to the EPA, Bioremediation is a “treatment that uses naturally occurring organisms to break down hazardous substances into less toxic or non-toxic substances”.

Bioremediation Techniques for Sewage Water TreatmentThe bioremediation can be done by using microorganisms, aquatic plants, earthworms, fish, plant extracts etc. Following are the various studies made to treat sewage water using different bioremediation techniques.

Use of microorganismsJain et al. (2013) reported that application of microbial consortium containing coenzymes (oxido-reductase group with proteolytic enzymes) and bacterial population belonging to hydrolytic, photosynthetic, flourscent, Bacillus sp., phosphate solubilizers, nitrifiers, denitrifiers, carbohydrate degraders, hydrocarbon degraders to the sewage water drains has resulted in reduction of pH, BOD and COD after 165 days (7.14, 24 mg lit-1 and 60 mg lit-1, respectively) at Bhamori region (Indore) as compared to raw sewage water (7.79, 290 mg lit-1 and 530 mg lit-1, respectively).

Use of aquatic plantsEl-Kheir et al. (2007) found that growing of duckweed (Lemna gibba) in sewage water for eight days has lowered TSS, BOD, COD, phosphate, ammonia, nitrate levels and Cryptophyceae phytoplankton population (14 mg lit-1, 30 mg lit-1, 88 mg lit-1, 6.20 mg lit-1, 2 mg lit-1, 0 mg lit-1 and 68.25 x 103 lit-1, respectively) in comparison with initial values (379 mg lit-1, 320 mg lit-

1, 800 mg lit-1, 11 mg lit-1, 10 mg lit-1, 8.32 mg lit-1 and 1337.5 x 103 lit-1, respectively).

Use of earthwormsSinha et al. (2008) reported that treatment of sewage water in a vermifilter kit having earthworms with a HRT of 1-2 hr has improved the pH and lowered the TSS, turbidity, BOD and COD of sewage water (7.05, 28 mg lit-1, 1.5 ntu, 1.97 mg lit-1 and 132 mg lit-

1, respectively) compared to vermifilter kit without earthworms (6.62, 116 mg lit-1, 3.6 ntu, 86.3 mg lit-1 and 245 mg lit-1, respectively) and raw sewage water (6.58, 390 mg lit-1, 112 ntu, 309 mg lit-1 and 293 mg lit-1, respectively).

Use of fishesSewage water treatment using duckweeds (Spirodela sp. And Lemna sp.) with a retention time of three days followed by fishes (Catla, Rohu, Mrigal, Silver carp and Silver barb) with a retention time of two days has resulted in improvement in pH, decrease in



total alkalinity, total hardness, TAN, NO2

--N, NO3

--N, PO

4--P, BOD, COD, TSS and total coliforms (7.2, 23%,

24%, 88%, 85%, 55%, 71%, 79%, 75%, 70% and 85.7%, respectively) compared to untreated sewage water (Jena et al., 2010).

Use of plant and animal extractsThere are several plant extracts developed for the sewage water treatment. Some of the examples are; Ecotan Series (Natural Based Coagulants) is a compound manufactured by Servyeco, Spain. It is a natural plant cationic coagulant derived from the Acacia (tannin compound) extracted from the bark of Acacia (Acacia mearnsii). It has strong coagulating action acting in colloidal systems in waste water, neutralizing the charges and bringing together the particles in suspension. It does not undergo hydrolysis in solution and its effectiveness is always optimal. It is liquid, ready to use, not requiring any dilution or mixtures and protects against corrosion of metallic parts. It is an organic based polymer, environmentally friendly (https://www.servyeco.com/uploads/catalogo/35_2_SERVYECO---NATURAL- COAGULANT. pdf - referred on 16th May, 2020).

Seeds of this tropical tree contain water-soluble, positively charged proteins that act as an effective coagulant for water and wastewater treatment. The study revealed that water treatment with Moringa stenopetala seed powder was found to be more effective for water purification than treatment with Moringa oleifera as a replacement coagulant (Abitu et al., 2018).

Chitosan is a one more coagulant which is a derivative of chitin present in the shell of shell fish, the exoskeleton of arthropods and the cell wall of yeast, fungi and yeasts. A new composite chitosan flocculant significantly improves the removal of chemical oxygen demand, suspended solids and aluminium ions from water (Defang and Jizu, 2004).

Use of Bioremediated Sewage WaterAl-Lahham et al. (2003) reported that irrigation of tomato (Variety: GS

12) with 100% treated sewage water

has recorded lower weight loss of tomato fruits after eight days, higher fruit diameter, total coliform, fecal coliform and total bacterial count on tomato skin (11%, 5.20 cm, 1.6 x 104, 3 x 102 and 189 x 102, respectively) in comparison with 100% potable water (14.6%, 4.05 cm, 340, 0 and 1 x 102, respectively).

Jamuna and Noorjahan (2009) reported that rearing of fishes for 60 days in Eichhornia sp. treated sewage water have recorded higher fish length, breadth and weight (8.4 cm, 3.6 cm and 10.4 gm, respectively) compared to raw sewage water (7.7 cm, 2.9 cm and 7.8 gm, respectively) and tap water (8.8 cm, 3.4 cm and 13.7 gm, respectively).

ConclusionThe bioremediation techniques have potential to treat the sewage water by using living organisms like microbes, algae, aquatic plants, earthworms and fishes.

However, the remediation capacity varies with the organism used and the content of sewage water. The bioremediated sewage water can be used for irrigation of field crops and fish cultivation etc. depend on the quality of treated sewage water. The techniques can be successful in practical sense only after standardization and testing of these techniques in large scale for sewage water treatment and after getting satisfactory results from the studies on the impact of bioremediated sewage water use on yield and quality of major field crops.

ReferencesAbitu, A., Yan, D., Girma, A., Song, X., Wang, H. and

E. Murat, 2018, Wastewater treatment potential of Moringa stenopetala over Moringa olifera as a natural coagulant, antimicrobial agent and heavy metal removals. Cogent Environmental Science, 4:1, DOI: 10.1080/23311843.2018.1433507

Al-Lahham, O., El-Assi, N. M. and Fayyad, M., 2003, Impact of treated wastewater irrigation on quality attributes and contamination of tomato fruit. Agril. Water Mgmt., 61(2003):51-62.

Bhardwaj, R. M., 2005, Status of wastewater generation and treatment in India. IWG-Env Joint Work Session on Water Statistics, Vienna, 20-22 June 2005.

Defang, Z. and Jizu, Y., 2004. Preparation and application of a new composite chitosan flocculant. International Journal of Environment and Pollution 21(5): 417-424.

El-Kheir, W. A., Ismail, G., El-Nour, F. A., Tawfik, T. and Hammad, D., 2007, Assessment of the efficacy of duckweed (Lemna gibba) in wastewater treatment. Int. J. Agri. Biol., 9(5):681-687.

Jain, S. K., Akolkar, A. B. and Choudhary, M., 2013, In-situ bioremediation for treatment of sewage flowing in natural drains. Int. J. Biotech. Food Sci., 1(3):56-64.

Jamuna, S. and Noorjahan, C. M., 2009, Treatment of sewage waste water using water hyacinth –Eichhornia sp. and its reuse for fish culture. Toxicol. Int., 16(2):103-106.

Jena, J. K., Patro, B., Patri, P., Khuntia, C. P., Tripathy, N. K., Sinha, S., Sarangi, N. and Ayyappan, S., 2010, Biological treatment of domestic sewage through duckweed-cum-fish culture: a pilot-scale study. Ind. J. Fish., 57(4):45-51.

Kaur, R., Wani, S. P., Singh, A. K. and Lal, K., 2013, Wastewater production, treatment and use in India. Water Technology Research Centre, New Delhi.

Sinha, R. K., Bharambe, G. and Chaudhari, U., 2008, Sewage treatment by vermifiltration with synchronous treatment of sludge by earthworms: a low-cost sustainable technology over conventional systems with potential for decentralization. The Environmentalist, 28(4):409-420.



EXTENSION EDUCATION AND RURAL DEVELOPMENT

20098

89. Stubble Burning: Causes, Consequences and AlternativesPAWAN KUMAR GAUTAM

Ph.D. Research Scholar, Dairy Extension Division, ICAR-National Dairy Research Institute, Karnal-132001 (Haryana) *Corresponding Author Email: [email protected]

IntroductionStubble burning refers to the deliberate act of setting the straw husk alighting once rice, wheat and alternative grains are harvested. The husk burning inside the regions of geographic area and Haryana was one in all the foremost reasons for the high levels of pollution in Delhi. The environmental impacts of the husk burning are tributary to the pollution levels and are choking the Indian cities like Delhi, that currently options a tag of one of the polluted cities of the planet. Some farmers realize it troublesome to have an effect on straw in traditional ways. As an example, a bumper crop will leave an implausible quantity of straw, which can be terribly troublesome to work into the soil or unfold equally across the world. Rainy weather once harvest will leave fields too wet until the next operation. Burning straw is taken into consideration a low-priced solution to the farmers. Under such circumstances, farmers might feel they have no alternative, however to burn the straw.

Causes1. The regions of north India are manufacturing two

crops on an outsized scale: rice and wheat, with wheat being planted and harvested within the dry winter season, and rice to coincide with the monsoon season.

2. The two cropping periods of Kharif and Rabi for rice and wheat, severally, have captive about to each other. This leaves concerning fifteen days in between the two crop cycles. This short span puts pressure on the farming community to arrange the farm for sequent crop throughout some days.

3. Burning the residue has become the most cost effective and fastest way to prepare the farm for succeeding crop cycle as a result of the HYV rice resulted within the residue that was a lot of taller than the basmati residue and at constant nowadays less eatable as fodder for animals.

4. Massive scale mechanisation promoted by the revolution has conjointly resulted during a condition that produces crop residue management more durable.

Consequences � Open burning of husk produces harmful smoke

that causes pollution. Open burning of husk is of incomplete combustion in nature. Hence an outsized quantity of harmful pollutants is emitted into the atmosphere.

� The heat from burning paddy straw penetrates one metric linear unit into the soil, elevating the temperature 33.8 to 42.2 degrees. This kills the microorganism and fungous populations crucial for fertile soil.

� Clouds of ash associate degreed smoke will travel over thousand kilometers and make an obstinate and non-clearing cloud. Smogginess fashioned of the smoke will increase the number of pollutants by manifolds inside the air, creating it troublesome to breathe.

� There was a big decrease of roughly five hundredth in sorptivity, final infiltration rate and hydraulic physical phenomenon within the burnt plots relative to the adjacent unburnt plots (Valzano et al., 1997).

� Stubble burning and tillage are two of the main processes chargeable for the decline of soil organic carbon concentration in crop soils, and therefore the ensuing soil degradation (Chan and Heenan, 2006).

AlternativesPossible alternatives to husk Burning will be:1. Providing husk grouping machines to farmers to

gather husk.2. Subsidizing or availing the husk grouping

machines at rent.3. Providing affordable labor to reap the paddy to

avoid husk generation. Providing labor would provide temporary employment to individuals.

4. Permitting animals to graze the husk.5. Decomposing husk within the farm field and

turning it into manure.6. Creating fodder for dairy animals out of collected

husk.7. Fixing Biomass fuel plants to come up with fuel

mistreatment paddy husk.8. The government ought to involve or invite

benefiting industries just like the trade to collaborate in husk/hull or husk assortment to use it proficiently.



9. Tempting packaging industries to gather husk to create packaging boxes that are additional atmosphere friendly than alternative non-disposable materials like thermocole and plastic.

ConclusionRice husk burning has been known as a serious environmental hazard. It is necessary to grasp the underlying causes and thus the prevailing things on why the farmers burn husk, then have an effect on the essential downside. Farmers left with few choices, however to burn the rice husk because of lack of labour throughout the harvest and limit the time offered for making ready the field for wheat cultivation, and therefore the burning is cheaper and needs less effort. Different efforts have to be compelled to be created

through Kisan cams, training, workshops for informing farmers concerning the choice usage of crop residues to beat the burning.

ReferencesChan, K.Y. and Heenan, D.P. (2006). The results of husk

burning and tillage on soil carbon sequestration and crop productivity in southeastern Australia. Soil Use and Management. 21(4): 427-431.

Valzano, F.P., Greene, R.S.B. and Murphy, B.W. (1997). Direct effects of husk burning on soil hydraulic and physical properties during a direct drill tillage system. Soil and Tillage analysis. 42(3): 209-219.

Yadav, R.S. (2019). Husk burning: a haul for the atmosphere, agriculture and humans. Down to Earth. Last Updated: Tues 04 June 2019.

20172

90. Agricultural Crisis and Lessons Amidst Corona Pandemic in India during Initial LockdownALOK K SAHOO AND TARAK C PANDA

SMS (Agricultural Extension), Scientist (Agricultural Engineering), KVK Bargarh, OUAT

Introduction: The varsities of Indian agriculture may not be confined to a standardised framework designed at top level policy makers which can be executed at grassroots with least prepared administration to face the crisis such as corona pandemic in the country, where food and nutritional security holds paramount significance to a second most populous nation. The agriculture which is the lifeline of every individual human being directly for survival should have been emphasized with proper guidelines for smoother agricultural and allied operations along with the concerns of stakeholders very before national lockdown announced. This might have saved the losses obtained from harvesting till tail end consumption of the farm produce resulting distress sale and uneven distribution. A sudden lockdown choked the transport system, labour availability and marketing of the farm produce which distorted the whole supply chain throughout the country.

Major Problems Faced in different Parts of the CountryIn Northern India: The Uttar Pradesh farmers were confronted with untimely rain and hailstorms in previous months prior to lockdown, the corona virus lockdown made them helpless for uncut standing matured wheat crop on field and uncertainty for lakhs of cane growers prepared with fields for sowing due to chaos on farm labourers movement restriction. The Madhya Pradesh wheat growers were in difficulty to harvest their produce due to absence of combine harvesters from Punjab and unavailability of manual labour. The Uttarakhand farmers suffered for

perishable items such as fruits and vegetables were dumped or wasted because of no potential buyers nearby and transports for distant regular market. The Punjab with a high colour for agriculture was not escaped from the crisis, as per report, unavailability of labourers for loading unloading of harvested wheat supplying to the Public distribution System and vegetable growers were able to sale the 20% of the whole produce owing to lack of demand from wholesaler, Hotels, Restaurants and caterers.

In Western India: The Rajasthan labour migration to their native state such as UP and Biihar troubled the harvesting of rabi crops such as wheat, mustard and barley, even though managed the same by local manpower but still not getting fair price at open market due to market not fully open and storage as a bigger problem. They were selling below Rs. 1600 per quintal of wheat at open market. The Maharashtra faced with severe challenge to Corona, farmers had no option but to dump their perishable fruits and vegetables high value produce such as grapes, citrus, Pomegranate, tomato, capsicum etc.

In Eastern India: The Odisha farmers were blessed with a bumper yield of vegetables and cash crops like pulses and oilseeds for a better income in hot summer but desperately looking for Government help to sell their produce at least in the local markets at whatever prices. There is a huge loss due to rotting of ripened vegetables 50-70% fall in the price in cities for distress sale of producers at farm gate. This was due to very restricted transport availability and exploitation of middle men for lowering the price in view of compulsion to sale by the producer. The Dairy farming



is hit similarly as lowering in demand from sweets shops, restaurants and road side caterers along with low consumption of value-added products among the city dwellers. The milk cooperative societies refused to purchase milk from the farmers as regularly procured due to no demand at upper head. The Chattisgarh mustard and pulse growers suffered severe crop damage due to untimely and heavy rainfall recently and unable to fix their crops as many of the labourers had fled. The West Bengal Potato growers were facing the challenge of stored potatoes in cold storages and godowns because of unavailability of labourers for loading unloading and truck drivers being scared of transporting which stopped intake of fresh harvested potatoes to the cold storage. The rice crop being a 70% share in total farm output from state would be severely affected due to disruption in preliminary preparation for next season.

In Southern India: The Andhra Pradesh Banana growers were expected to harvest 27 lakh metric tonnes from which 70% were meant for other states with Big cities in Northern India. Almost all the markets were closed which stopped the supply chain, the loanee farmers were not in a position to repay. In Telengana, the vegetables such as tomatoes were sold at very low price @ Rs 6 per kg and some places sweet lime were thrown on roads due to no transport facilities. The Karnataka floriculture business was clogged rapidly as places of worship closed and ban on all cultural, traditional or religious ceremonies all over India and subsequently no demand for flowers caused a huge loss to a highly perishable enterprise. With lockdown, the plantation crops industries were severely hit with restriction on export and domestic consumption. Tamilnadu Banana and flower farmers had similar experience in selling their produce at timely sales. Farmers left the banana bunches to ripen in trees because of transport restrictions to most exporting nearby state Kerala. In Kerala, the pineapple sale was 20 tons per daily average as compared to 1200 tons of daily average production piling up the stocks in the market. Similarly, the daily average mango procurement production ratio was 1:66 in the state. Anonymous (2020)

Lessons learnt and way forward: The unprecedented lockdown which seized all sorts of agricultural supply chain and economy has created a hasty scenario of uncertainties and unpreparedness to such a crisis. Despite a lot of limitations aligned with the farming sector underestimated prior to lockdown were notably realized by the stakeholders during crisis. The marketing of produce was the biggest problem during crisis mostly perishable items such as fruits, vegetables, flowers, milk and milk products. This is alarming for development of infrastructure such as more cold storage and cold supply chain for perishable commodities like NDDB AMUL model. This horticultural sector should be incorporated in Minimum Support Price like assurance support scheme to save producers from distress sales and facilitate them technology cum value addition and market led farming with high quality and standards for export

and premium sale in domestic market. The revamped marketing system to APMC mandi system “one nation one market” would integrate and digitize the domestic market and prevent farmers from restricted sale at a particular mandi to a particular trader or middle men as per the regulations. The e-NAM platform would be beneficial for farmers as they can sell their produce with uploading photograph to the online bidders with different rate chart as per quality from a remote location maintaining social distancing and without physical transportation of real produce to the market. The food and nutritional security is another challenge for supplying food grains with pulse and nutri-cereals to the migrant labourers in cities and reverse migrated people at their villages. The Public distribution System such as FCI needs to relook through direct procurement from farmers or mandis at block or panchayat level and distributing the same to the local people without storing in godown which saves time and cost and unnecessary repeated function facilitating a speedy action for both the producers and consumers. The MGNREGS programme should include farming activities and skilled, semi-skilled farming operations along with unskilled work for both the gender with assured livelihood support to the jobless migrants at villages. The agricultural practices should be encouraged with social distancing and wearing mask or cloth among the farming communities. The home sanitation and covid-19 regulations are to be facilitated by agro-advisories through ICT media and real field demonstration by extension professionals. The Farmer Producers Company or Farmers organisation and their leaders can participate through online video conferencing platform to the KVK, line department officials and marketing system for real time information access and correct decision in farm activities.

ReferenceAnonymous (2020). Across India, a massive

agricultural crisis in the making due to coronavirus shutdown. The New Indian Express. Published: 12th April 2020 11:31 AM



20205

91. Role of Eco-Friendly Agricultural Practices used by Tribal Farmers in Crop Production for Sustainable DevelopmentSONAM UPADHYAY

Department of Agricultural Extension, JNKVV, Jabalpur *Corresponding Author Email: [email protected]

To a outstanding quantity, future meals security and economic independence of growing nations would rely on improving the productiveness of bio-physical assets via the utility of sustainable manufacturing strategies, via improving tolerance of plants to unfavorable environmental conditions and by way of lowering crop and publish-harvest losses caused by pest and illnesses. Indigenous agricultural practices can play a key position inside the design of sustainable and green agricultural systems, increasing the probability that the agricultural population will accept, increase and maintain innovations and interventions. Green and environmentally pleasant are synonyms used to consult items and services considered to inflict minimum or no harm at the environment. Tribal’s commonly undertake their traditional strategies of farming which they get from their history and this farming network that’s because of their low financial and cognizance fame generally adopts low price farming practices. They deal with land as a mother so, they emotionally in addition to culturally connected with nature. So, it becomes necessary to have a better recognition approximately their volume of belief regarding green farming practices. The contemporary agriculture has been a success in meeting the accelerated meals needs of alarmingly developing populace. However, the trouble associated with modern agriculture like, the high fee of inorganic or chemical fertilizers and plant protection chemical compounds, stagnated yield degrees inside the recent years and the mounting health and environmental risks have compelled many farmers and scientists to consciousness interest on ecologically sound feasible and sustainable farming. To be able to mitigate those fitness dangers and convey out herbal balance and safety of ecosystem, natural motion has commenced in numerous elements of the arena, wherein no chemical fertilizers and plant safety chemicals are used in the cultivation of discipline vegetation, vegetables and culmination. Its miles ascertained that the indiscriminate use of agro-chemical substances and pesticides purpose adverse adjustments in the ecological balance. As a result, understanding the significance of green practices of farming structures which can be environmentally sound, profitable manufacturing and preserve the social fabric of the agricultural network, this observe changed into undertaken to establish and decorate rural surroundings and agricultural practices.

HistoryThe eco-friendly agriculture movement was first recognized internationally in a joint study of the World Union and Future Harvest Foundation published in 2001 called “Common Ground, common Future”. The report was later expanded to become a book called “Eco-agriculture: Strategies to Feed the World and Save Wild Diversity”. (McNeely and Scherr 2001). Eco-friendly and environmentally friendly are synonyms used to refer to goods and services considered to inflict minimum or no harm on the environment. To make consumers aware environmentally friendly goods and services often are marked with eco-labels. Eco-friendly farming is the process of producing food naturally. This method avoids the use of synthetic chemicals and generally modified organisms to influence the growth of crops.

Aim of Eco-Friendly PracticesThe aim of eco-friendly agriculture is to manage the resources of rural communities to improve their welfare, Preserve biodiversity and ecosystem services, and develop more productive and sustainable farming system. Eco-friendly and environmentally friendly are synonyms used to refer to goods and services considered to inflict minimum or no harm on the environment. To make consumers aware environmentally friendly goods and services often are marked with eco-labels. Eco-friendly farming is the process of producing food naturally. This method avoids the use of synthetic chemicals and generally modified organisms to influence the growth of crops. This involves musing techniques to achieve good crop yields without harming the natural environment or the people who live and work in it. Eco-friendly agriculture, now emerging as a holistic approach to ecologically and socially responsible land use, represents a vision of rural communities managing their landscape and resources to jointly achieve three goals:

� Enhance rural livelihood � Conserve or enhance biodiversity and eco-system

services � Develop more sustainable and productive

agricultural system

Eco-Friendly Agricultural Practices � Agronomical Practices



– Early sowing– Nursery raising– Plant spacing– Hand weeding

� Seed management– Selection of suitable variety– Seed treatment– Grading of seed

� Soil management– Conservation tillage– Bunding

� Water management– Proper drainage– Selection of crop

� Use of Bio-fertilizer– Azolla– Azotobactor

� Integrated Nutrient management– FYM– Vermicompost– Green manure– Balanced dose of fertilizer

� Integrated Insect and Disease management– Use of light trap– Use of pheromone trap– Control insect by predators

� Storage– Storage at proper moisture– Storage structure– Use of treated bags

Eco-Friendly Approaches for Farming System � Organic farming: It included farming device

that strives for sustainability, the enhancement of soil fertility and biological diversity whilst, with uncommon exceptions, prohibiting artificial pesticides, antibiotics, artificial fertilizers, usually modified organisms boom hormones. Natural farming is strategies, which involves cultivation of plants and rearing of animals in herbal methods. This method involves the usage of biological materials, avoiding synthetic substance to preserve soil fertility and ecological stability thereby minimizing pollution and wastage.

� Biological farming: Biological farming is a chemical unfastened approach of farming that target improving the microbiology as a manner of increasing plant growth and produce yield. Biological farming includes (however is not constrained to): natural farming. Biological farming is using modern-day generation and new methods, but makes use of simplest those that don’t intervene with natural systems and do no longer reason damage down the road.

� Nature farming: Nature farming is a holistic approach wherein farmers are discouraged to buy market primarily based inputs like chemical fertilizers, chemical insecticides and many others. For growing flora within zero price range

and endorsed to grow healthy soil with pleasant earthworms and thereby develop healthy plants. The ideas of nature farming utilize and adopts crop manufacturing to comply to these dynamic and balanced manufacturing systems in nature, which are a end result of the interactions of daylight, water, soil, animals, flowers, and microorganisms in herbal ecosystems. It is very vital to take a look at nature without being too confident of our expertise however with a modest, clean and pure kingdom of mind. Similarly, growing correct plants requires the improvement of love to the crops. Most effective with such love, a farmer can understand the requirements of soil and for crops to develop healthily, and hence carry out important control practices.

� Regenerative agriculture: Regenerative agriculture” describes farming and grazing practices that, amongst other blessings, opposite climate change by rebuilding soil natural count number and restoring degraded soil biodiversity – ensuing in each carbon drawdown and improving the water cycle. The key to regenerative agriculture is that it now not only “does no harm” to the land however honestly improves it, the usage of technology that regenerate and revitalize the soil and the environment. Regenerative agriculture leads to healthful soil, able to generating excessive great, nutrient dense meals even as simultaneously improving, instead of degrading land, and in the end leading to productive farms and healthy communities and economies. It’s far a dynamic and holistic, incorporating permaculture and natural farming practices, including conservation tillage, cover crops, crop rotation, composting, cellular animal shelters and pasture cropping, to increase food production, farmers’ income and in particular, topsoil.

� Permaculture: “Permaculture, firstly ‘permanent agriculture’, is frequently viewed as a hard and fast of gardening strategies, but it has in reality advanced into an entire design philosophy, and for some people a philosophy for lifestyles. Its central subject matter is the introduction of human systems which provide for human wishes, however the use of many natural elements and drawing idea from natural ecosystems. Its goals and priorities coincide with what many people see as the middle requirements for sustainability.”

To Boost Agriculture DevelopmentAgriculture is an activity directly related to the use of natural resources. This is compounded by farming practices that pay little heed to the rules of ecosystem balance and environmental conservation, which will in turn have an impact on agriculture itself.

� Benefit local farming communities � Develop habitat networks in non-farmed areas, � Reduce land conversation to agriculture by

increasing farm productivity � Minimize agricultural pollution



� Modify management of soil � Water and vegetation resources

ConclusionIn a healthy farming system, agriculture works in harmony with the natural environment. This started out with wholesome soil that stores water and vitamins and provides a solid base to help plant roots. In a sustainable system, soil; is saved in balance. Crops are

circled thru the fields to replace nutrients in soil. In which there’s cattle, animal gaze the land, then waste from those animals is used to fertilize the soil. The idea is that as farmer take from the land in addition they supply again.

Natural, mechanical, bodily, and cultural practices of agriculture utilized in ecological agriculture. Due to the fact 1985 DAE has been running closer to development of this opportunity strategy and termed it as “green agriculture”.

HOME SCIENCE

20161

92. Sleeping Ergonomics: What Sleeping Position are You??DIVYA MARTOLIA

Research Scholar, Dept of Family Resource Management, College of Community Science, Punjab Agricultural University, Ludhiana, Punjab, India. *Corresponding Author Email: [email protected]

Sleep has major impact on our health. After the end of the day our body needs to recharge the energy therefore, it is important to have proper sleep with healthy sleeping posture. The improper sleep positions and mattresses that do not provide proper support or pressure point relief can cause physical pain, numbness and long-term body ache. People have a habit of changing position on the back, on either side or on the stomach multiple times while sleeping in bed. A healthy sleeping posture reduces the stress from whole body, also prevent problems from developing, and stop reoccurrence. The good posture and better selection of sleeping mattress and pillow has huge impact on the quality of sleep as this contributes to sleep ergonomics.

Sleeping postures -pros and cons

Sleeping posture Pros Cons

Back sleeping

This position has less chance of back pain.Enhance good digestion.Helps prevent wrinkles and acne by keeping your face off the pillow, out of contact with bacteria.

May perpetuate snoring because gravity causes the tongue or loose tissue in the throat to collapse and block the airway.

Sleeping posture Pros Cons

Side sleeping

Eases heartburn and acid refluxDiminishes sleep apnea symptomsImproves blood flow to and from the heart.

The side-lying fetal position can cause back and neck pain.Cuts off circulation in the arm resting during side sleep because of weight on the arm for an extended time.

Stomach sleeping

Diminishes sleep apnea symptoms.

This position strains the back, neck, and joints due to keeping the head and neck twisted to one side for an extended time.May perpetuate the discomfort of acid refluxThis position puts pressure on certain organs for an extended time

Source: John-Manuel Andriote

Suggestion for Selection of Mattress and Pillow

Mattress selection � The mattress designed keep in mind the spine

in proper position while sleeping, which is conventional to spine natural curves.

� The mattress designed in such a way that it evenly distributes the pressure across the body for proper blood flow, furthermore improve sleep quality.

� Select the mattress designed with the proper



edging.

Pillow selection � Conventional to spine natural curves. � It should be in proper shape, curve and fit with the

user sleeping posture. � It should support unlike sleeping positions such

as side sleeper, back sleeper, stomach sleeper. � It should give support to the head of the user. � Provide soft touch. � It should reduce stress points.

� It should have better facial air circulation.Conclusion: The poor sleeping posture and the

bad choices of mattress and pillow for sleeping founds to be uncomfortable sleep nights. Consequently, sleep ergonomic is an imperative approach to improve the quality of sleep, decrease the physical pain and numbness of body. Awareness of sleep ergonomics indeed helps the people to improve the sleep posture, and this let people to wake up with fresh mind and more energy for the following day task.

ReferenceMattresses Matter: Ergonomic Guidelines How to

Sleep Soundly Written by Alan Hedge, PhD, CPE; Reviewed by Richard D. Guyer, MD 2017 sept17 www.spineuniverse.com › Wellness › Sleep

The Pros and Cons of Back, Side, and Stomach SleepingBy John-Manuel Andriote November 28, 2019 www.

saatva.com › blog › sleeping-positions-pros-cons



FOODS AND NUTRITION

20132

93. Value Added Dairy OptionsSTEPHY DAS AND DR. MANJU K. P.

Subject Matter Specialists, ICAR Krishi Vigyan Kendra, Kannur

Dairy farmers in India are now staring at an uncertain future after the outbreak of Covid-19. Milma reduce the daily procurement from farmers so that tons of milk are thrown away. Many household and farm family’s consumption shot up due to the unawareness of processing milk-based products. In this context, processing methods of consumer preferred products are given below.

PaneerIndian cheese, called paneer or chenna when combined with sugar is an unripened cheese made by coagulating milk with lemon juice, then leaving to drain to allow the curds and whey to seperate it is then pressed into blocks.

� 3 litres milk � 6 tablespoons strained lemon juice and vinegar

For the chenna � 1 teaspoon caster sugar � 1 teaspoon maida or plain flour

TO MAKE the paneer, pour the milk into a large heavy based saucepan. Bring to the boil, stirring with a wooden spoon so that the milk doesn’t stick to the base of the pan. Reduce the heat and stir in the lemon juice, then heat over low heat for a few more seconds before turning the heat off as large bits of curd start to form and release the yellow whey.

IF the curds are slow to form, put the pan over low heat again for a few seconds. This helps with the coagulation.

LINE a colander with muslin or cheese cloth so that it overlaps the sides. Pour off the whey, collecting the curds gently in the colander. Gently pull up the corners of the cheese cloth so that it hangs like a bag, twist the cloth so that the whey is released, then hold the bag under running water to wash off the remaining whey, twisting some more to remove the excess liquid.

LEAVE the bag to hang from your tap for several hours so the weight of the curd releases more liquid and the cheese compacts. To remove more liquid, press the bag under a heavy weight such as a tray with some tinned food piled on top, for about 1 hour. This will form a firm block of paneer. When the block is firm enough to cut into cubes, the paneer is ready for use.

TO MAKE chenna, remove the cheese from the bag and knead the paneer well with the palms of your hands until it is very smooth. Combine the panner with

the sugar and maida, Kneading the sugar in until it is fully incorporated.

Kulfiyoung and old take great delight in these flavoured ices which are sold in India

� 2 litres milk � 10 cardamom pods, lightly crushed � 6 tablespoons sugar � 15 g almonds, blanched and finely chopped � 15 g unsalted pistachio nuts

PUT the milk and cardamom pods in a heavy based saucepan and bring to the boil. Reduce the heat to low and simmer, stirring frequently, for about 2 hours, until the milk has reduced to a third of the original amount, about 750 ml. Whenever a thin skin forms on top, stir it back in.

ADD the sugar to the pan, simmer for 5 minutes, then strain into a shallow plastic freezer box. Add the almonds and half the pistachios, then cool. Put twelve 75 ml kulfi moulds or dariole moulds in the freezer to chill.

PLACE the kulfi mixture in the freezer and every 20 minutes, using electric beaters or a fork, give the ice cream a good stir to break up the ice crystals. When the mixture is quite stiff, divide it among the moulds and freeze until hardened completely. Dip the moulds in hot water and turn out the kulfi, sprinkle with the remaining pistachios.

ShrikhandThis is yet another milk-based recipe. It is made by straining yogurt and flavouring it with saffron and cardamom to give a thick, rich, creamy dessert.

� I/2 teaspoon saffron strands � 3 cardamom pods � 275 ml thick natural yogurt � 3 tablespoon caster sugar � A few toasted flaked almonds

SOAK the saffron in 1 teaspoon boiling water. Remove the cardamom seeds from the pods and coarsely crush them in a spice grinder or pestle and mortar.

PUT the yogurt, sugar, cardamom and saffron in a bowl and beat until well mixed. Divide among four bowls and refrigerate before serving. Serve with toasted almonds sprinkled on top.

Volume XIX Issue No. 01 01 June 2020 Pages: 148 - Agrobios ...

Documents

Transcript of Volume XIX Issue No. 01 01 June 2020 Pages: 148 - Agrobios ...