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Re: Gesunde Pflanzen (10.1007/s10343-012-0286-x)Ameliorative Effects of Brassinosteroids on Improving Growth and Productivity of Snap Beans Grown Under High Temperature Stress

A. El-Bassiony A. Ghoname M. El-Awadi Z. Fawzy N. Gruda

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ArticleTitle Ameliorative Effects of Brassinosteroids on Improving Growth and Productivity of Snap Beans Grown Under High Temperature Stress

Article CopyRight - Year Springer-Verlag Berlin Heidelberg 2012

Journal Name Gesunde Pflanzen

Corresponding Author Family Name Ghoname

Particle

Given Name A. A.

Suffix

Organization National Research Center

Address Cairo, Dokki, Egypt

Division Department of Vegetable Research

Email ghoname1@yahoo.com

Author Family Name El-Bassiony

Particle

Given Name A. M.

Suffix

Organization National Research Center

Address Cairo, Dokki, Egypt

Division Department of Vegetable Research

Author Family Name El-Awadi

Particle

Given Name M. E.

Suffix

Organization National Research Centre

Address Cairo, Dokki, Egypt

Division Botany Department

Author Family Name Fawzy

Particle

Given Name Z. F.

Suffix

Organization National Research Center

Address Cairo, Dokki, Egypt

Division Department of Vegetable Research

Author Family Name Gruda

Particle

Given Name N.

Suffix

Organization University of Bonn

Address Bonn, Germany

Division Faculty of Agriculture

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Received 1 October 2012

Schedule Revised

Accepted 18 October 2012

Abstract The effect of brassinosteroids (BRs) was tested, in order to assess the possibility of alleviation of the adverse effects of high temperature stress on snap bean plants during delayed summer cultivation. Therefore, two field experiments were carried out in successive seasons, 2010 and 2011, with two bean cultivars ‘Paulesta’ and ‘Oxzira’, spraying with BRs of the following concentrations 0 (control), 25, 50 and 100 ppm. Plant growth, yield and pods quality of beans were studied. Spraying bean plants with BRs at a concentration of 25 and 50 ppm increased vegetative growth, total yield and quality of pods with no significant difference between both treatments. Using BRs at 25 ppm increased the total free amino acids (FAA) in leaves and total phenolic acids in the pod in comparison to control-treatment. ‘Oxzira’ cultivar resulted in the highest number of leaves, number of branches, dry weight of whole plant, total yield, total FAA in leaves and total phenolic acids in pod. Whereas, ‘Paulesta’ cultivar had the highest of plant length and total FAA in pods.In der vorliegenden Arbeit wurde untersucht, inwiefern Brassinosteroide (BRs) den oxidativen Stress durch hohe Temperatureinwirkungen bei Brechbohnen während eines verzögerten Sommeranbaus verringern. Dazu wurden zwei Feldversuche mit den zwei Bohnensorten ‘Paulesta’ und ‘Oxzira’ durchgeführt. Besprüht wurden die Pflanzen mit BRs in folgenden Konzentrationen 0 (Kontrolle), 25, 50 und 100 ppm in den Jahren 2010 und 2011. Pflanzenwachstum, Ertrag und Qualität der Brechbohnen wurden untersucht. Das Besprühen der Bohnenpflanzen mit BRs bei einer Konzentration von 25 und 50 ppm erhöhte das vegetative Wachstum, den Gesamtertrag und die Qualität der Hülsen. Es konnte kein signifikanter Unterschied zwischen den beiden Behandlungen festgestellt werden. Bei der Verwendung von einer Konzentration von 25 ppm erhöhten sich – im Vergleich zur Kontrolle – die gesamten freien Aminosäuren (FAA) in den Blättern sowie der Phenolsäuregehalt in den Hülsen. Die Sorte ‘Oxzira’ zeigte die höchste Anzahl an Blättern und Verzweigungen, Trockengewicht, Gesamtertrag, FAA in Blättern und Phenolsäure in den Hülsen. Die Sorte ‘Paulesta’ hatte dagegen die größte Pflanzenhöhe und FAA in den Hülsen.

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ORigiNal aRTiClE

Abstract The effect of brassinosteroids (BRs) was tested, in order to assess the possibility of alleviation of the ad-verse effects of high temperature stress on snap bean plants during delayed summer cultivation. Therefore, two field experiments were carried out in successive seasons, 2010 and 2011, with two bean cultivars ‘Paulesta’ and ‘Oxzi-ra’, spraying with BRs of the following concentrations 0 (control), 25, 50 and 100 ppm. Plant growth, yield and pods quality of beans were studied. Spraying bean plants with BRs at a concentration of 25 and 50 ppm increased vegetative growth, total yield and quality of pods with no significant difference between both treatments. Using BRs at 25 ppm increased the total free amino acids (Faa) in leaves and total phenolic acids in the pod in comparison to control-treatment. ‘Oxzira’ cultivar resulted in the high-est number of leaves, number of branches, dry weight of whole plant, total yield, total Faa in leaves and total phe-nolic acids in pod. Whereas, ‘Paulesta’ cultivar had the highest of plant length and total Faa in pods.

Zusammenfassung in der vorliegenden arbeit wurde untersucht, inwiefern Brassinosteroide (BRs) den oxi-dativen Stress durch hohe Temperatureinwirkungen bei Brechbohnen während eines verzögerten Sommeranbaus verringern. Dazu wurden zwei Feldversuche mit den zwei

Bohnensorten ‘Paulesta’ und ‘Oxzira’ durchgeführt. Be-sprüht wurden die Pflanzen mit BRs in folgenden Konzen-trationen 0 (Kontrolle), 25, 50 und 100 ppm in den Jahren 2010 und 2011. Pflanzenwachstum, Ertrag und Qualität der Brechbohnen wurden untersucht. Das Besprühen der Bohnenpflanzen mit BRs bei einer Konzentration von 25 und 50 ppm erhöhte das vegetative Wachstum, den ge-samtertrag und die Qualität der Hülsen. Es konnte kein signifikanter Unterschied zwischen den beiden Behand-lungen festgestellt werden. Bei der Verwendung von einer Konzentration von 25 ppm erhöhten sich – im Vergleich zur Kontrolle – die gesamten freien aminosäuren (Faa) in den Blättern sowie der Phenolsäuregehalt in den Hülsen. Die Sorte ‘Oxzira’ zeigte die höchste anzahl an Blättern und Verzweigungen, Trockengewicht, gesamtertrag, Faa in Blättern und Phenolsäure in den Hülsen. Die Sorte ‘Pau-lesta’ hatte dagegen die größte Pflanzenhöhe und Faa in den Hülsen.

Introduction

Snap bean (Phaseolus vulgaris) has a great importance as one of most vegetable export crops in Egypt. Therefore, expansion in cultivation of snap beans is growing rapidly and could be expanded until the early summer. However, bean plants are relatively sensitive to high temperature stress that may occur in the field, especially when grown under delayed sowing, e.g. in april and May, which nega-tively affects its growth, yield and even the quality of pods.

The yield of snap beans is severely reduced under high temperature condition (Prasad et al. 2002). according to Konsens et al. (1991) and Monterroso and Wien (1990) high temperatures during the reproductive phase caused a reduc-tion in pod and fruit set in green beans, due to enhanced

gesunde PflanzenDOi 10.1007/s10343-012-0286-x

Ameliorative Effects of Brassinosteroids on Improving Growth and Productivity of Snap Beans Grown Under High Temperature Stress

Abdelmohsin Mahmoud El-Bassiony · Abdalla Abdel Aziz Ghoname · M. E. El-Awadi · Z. F. Fawzy · N. Gruda

a. a. a. ghoname () · a. M. El-Bassiony · Z. F. FawzyVegetable Research Department, National Research Center, Dokki, Cairo, Egypt e-mail: ghoname1@yahoo.com

M. E. El-awadiBotany Department, National Research Centre, Dokki, Cairo, Egypt

N. grudaFaculty of agriculture, University of Bonn, Bonn, germany

Received: 1 October 2012 / accepted: 18 October 2012© Springer-Verlag Berlin Heidelberg 2012

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abscission of flower buds, flowers, and pods. also, Omae et al. (2006) stated that high temperature (30 °C/26 °C), at the onset of flowering, negatively affected both number of pods/plant and individual pod weight.

BRs are a new group of plant hormones that are isolated from the extract of Brassica napus pollen grains (grove et al. 1979). Since this report in “Nature”, they are still suc-cessfully used as a natural product with different objec-tives in plant cultivation. BRs have been found to influence diverse physiological processes and promote stem elonga-tion and cell division (Mitchell et al. 1970). Krishna (2003) reported that BRs also play a significant role in the ame-lioration of various biotic and abiotic stresses. according to Zhu et al. (1998), for instance, an exogenous application of BRs could alleviate the adverse effects of heat stress on rice. a beneficial effect of BRs foliar spraying on the yield of tomatoes, cultivated under heat stress was reported from Singh and Shono (2005), mainly due to an increase in fruit numbers. Furthermore, Clouse and Sasse (1998) stated a positive relation of naturally occurring BRs and stimulation of pollen tube growth.

The role of BRs, as a kind of nontoxic, natural, and envi-ronmentally friendly hormone for enhancing stress toler-ance may have a promising prospect especially for snap beans grown under hot conditions, due to delayed cultiva-tion in summer. However, only limited studies have been reported so far on the role of BRs in the thermotolerance of bean plants.

The objective of the present study is to investigate the effect of BRs on improving growth, productivity and qual-ity of two snap bean cultivars grown under high temperature stress, due to delayed summer cultivation.

Materials and Methods

location and growth Conditions

The field experiment was conducted at the agricultural Research Station of National Research Center in the Nubaria region, which is located in the North West of the Nile delta in Egypt, 47 km South of alexandria at an altitude of 164 m above mean sea level (latitude of 30°30′N and longitude of 30°20′E). The location climate is dry with non-effective rainfall amount of 16.5 mm, occurring throughout the winter season. The climatic parameters were recorded from the Nubaria meteorological station. Monthly tempera-tures (max and min) and relative humidity recorded during the two growth seasons are summarized in Table 1.

The experimental soil had a sandy texture with a pH of 7.6, EC-value of 0.81 dS m− 1, and the organic matter of 0.19 % with a content of 15.00, 9.40, 16.00 mg per 100 g soil of N, P, and K, respectively. Normal agricultural prac-

tices required for snap bean production were applied as commonly followed in the farm and as recommended by the Egyptian Ministry of agriculture.

Plant Material and Treatments

The BR analogues used in the present work are β-sitosterol (22,23-Dihydrostigmasterol, (stigmasta-5-en-β-ol,) pur-chased from Mallinckrodt (Paris, KY, USa). Seeds of two snap bean (Phaseolus vulgaris l.) cultivars ‘Paulesta’ and ‘Oxzira’ were sown in the last week of april in 2010 and 2011, respectively. Seeds were sown on two sides of the ridge 10 cm apart; ridge was 80 cm width and 4 m length. Each plot included four ridges and the plot area was 12.8 m2. Thirty days old bean plants were sprayed twice in 15 days intervals with BRs of three concentrations 25, 50 and 100 ppm. in addition, a control treatment, water only, was used.

Sampling and Data Recording

Vegetative Growth

a random sample of five plants from each plot was taken 45 days after sowing (DaS). The following vegetative charac-ters were recorded: plant length (cm), number of leaves and number of branches as well as fresh and dry weight of whole plants (gram per plant).

Pods Yield at the harvest stage the mature pods of beans for each experimental plot were collected along the harvesting season and total pods yield was recorded in kilogram per square meter.

Pods Quality a random sample of 50 pods from each plot was taken and the physical properties were recorded, including average pod weight (gram), and pod length (centimeter). in addition, protein percentage was determined according to aOaC (1990). Total Faa (milligram per gram fresh weight) were determined using the ninhydrin colorimetric method, defined by Plummer (1978). The colorimetric method of Folin-Denis as described by Daniel

Table 1 average monthly maximum (Tmax) and minimum (Tmin) tem-perature and relative humidity, during the two studied seasonsMonth 2010 2011

Tmax (°C)

Tmin (°C)

RH (%)

Tmax (°C)

Tmin (°C)

RH (%)

april 28.3 14.2 61 28.6 14.8 59May 32.1 17.4 61 32.4 17.5 60June 34.7 20.2 63 34.8 20.4 64July 34.9 21.8 67 35.2 22.3 67

5758596061626364656667686970717273747576777879808182838485868788

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and george (1972) was used for the chemical determination of total phenolic acid (milligram per gram dry weight).

Experimental Design and Statistical Analysis

The treatments (BRs and cvs.) were arranged in a split plot design with four replicates, where bean cultivars were arranged in main plots and BRs foliar application treat-ments in the sub plots. The obtained data were statistically analyzed and means separation was carried out using least Significant Difference (lSD) at P < 0.05 according to the method described by gomez and gomez (1984).

Results

Effect of BRs Foliar application and Cultivars on Vegetative growth of Snap Bean Plant

Effect of BRs

Spraying snap bean plants with BRs at 25 and 50 ppm increased plant length, number of leaves and shoots per plants, number of branches per plants as well as fresh and dry weight of leaves (Table 2), with no significant differ-

ence between 25 and 100 ppm. On the contrary, the lowest number of leaves and shoots per plant, number of branches per plants and fresh and dry weight of leaves were recorded in unsprayed plants.

Effect of Cultivars

Table 2 shows that ‘Oxzira’ cultivar was very vigorous and had the highest number of leaves, number of branches as well as fresh and dry weight of whole plant in the two sea-sons. Except for plant length, the lowest vegetative growth parameters were obtained with the ‘Paulesta’ cultivar. These results were consistent in both seasons.

Effect of the Interaction

The interaction between BRs and cultivars had a significant effect on vegetative growth characteristics expressed as plant length, number of leaves and fresh and dry weight of whole plant (Table 2). However, the highest growth of snap bean plants was found using ‘Oxira’ cultivar when sprayed with BRs of 25 and 50 ppm in both seasons, except for plant length which was highest with the ‘Paulesta’ cultivar, sprayed with BRs at 25 ppm. The lowest growth parameters were found in the control treatment and ‘Paulesta’ cultivar. These results were consistent in both seasons.

Table 2 Effect of cultivars and brassinosteroids foliar application on vegetative growth parameters of bean plantsTreatments Plant length (cm) leaf number Branch number leaves fresh weight (g) leaves dry weight (g)

2010 2011 2010 2011 2010 2011 2010 2011 2010 2011Effect of BRsControl 38.93 42.06 16.25 17.48 9.75 10.40 30.30 30.50 6.71 6.9025 ppm 44.25 46.87 24.75 26.25 12.50 12.90 50.68 50.71 8.69 8.7250 ppm 41.13 43.96 28.50 30.84 11.75 12.08 53.80 54.11 8.61 8.64100 ppm 42.45 45.90 17.50 18.85 13.75 14.26 28.63 28.67 5.29 5.60LSD at 5 % 2.65 3.02 4.92 5.13 5.45 5.71 11.35 9.84 1.67 2.65Effect of cultivarsPaulesta 44.04 47.38 18.88 20.34 11.38 12.17 36.46 37.21 6.89 6.70Oxzira 39.34 42.01 24.63 26.36 12.50 12.65 45.24 44.78 8.03 7.95LSD at 5 % 2.65 3.02 3.06 3.27 NS NS 4.81 2.71 1.80 0.42Effect of the interactionControl Paulesta 41.25 44.09 14.00 15.03 8.50 9.04 29.55 29.68 41.25 6.13

Oxzira 36.60 40.02 18.50 19.94 11.00 11.76 31.05 31.31 7.42 7.2825 ppm Paulesta 46.00 49.27 20.00 21.57 12.00 12.85 39.25 40.25 8.36 8.30

Oxzira 42.50 44.46 29.50 30.93 13.00 12.94 62.10 61.16 9.08 9.0850 ppm Paulesta 42.25 46.18 24.50 26.48 12.00 12.85 45.35 46.90 7.34 7.19

Oxzira 40.00 41.73 32.50 35.20 11.50 11.31 62.25 61.32 9.95 10.03100 ppm Paulesta 46.65 49.98 17.00 18.30 13.00 13.94 31.70 32.02 5.50 5.19

Oxzira 38.25 41.82 18.00 19.39 14.50 14.58 25.55 25.32 5.69 5.39LSD at 5 % 2.65 3.02 10.60 11.33 NS NS 7.35 6.78 4.36 3.95

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Effect of BRs Foliar application and Cultivars on Pod Yield

Effect of BRs

BRs foliar spraying significantly increased pod weight and total pod yield in comparison to unsprayed plants in the two seasons. Moreover, total pod yield was highest at concentra-tions of 25 and 50 ppm. However, spraying snap bean plants with BRs at 50 and100 ppm had a positive effect on pod weight. Whereas, pod length responded insignificantly and failed to reach a level of 5 % significance (Table 3). Total pod Yield was higher with concentrations of 25 ppm, while pod weight and length was not. Such results seem to be a function of increased number of pods which increases total pod yield.

Effect Cultivars

Table 3 shows that the highest pod length, pod weight and total pod yield were recorded with the ‘Oxira’ cultivar. No significant differences were shown for pod length and pod weight in comparison to the ‘Paulesta’ cultivar in both seasons.

Effect of the Interaction

The interaction between cultivars and BRs had a signif-icant effect only on the total yield of snap beans in both seasons. The highest total pod yield was recorded using BRs of 50 ppm with the ‘Oxira’ cultivar in the two seasons (Table 3).

Effect of BRs Foliar application and Cultivars on Pod Quality

Effect of BRs

Tables 4 and 5 show that the nutritional value of snap bean (nitrogen content, phenolic acid and protein) pods were found to be higher with BRs of 25 ppm in the two observed seasons, while the highest phosphorus and potassium per-centages in pods were recorded with BRs at 50 ppm. More-over, the highest Faa in pods were found using foliar spray of BRs of 100 ppm in both seasons.

Effect of Cultivars

Tables 4 and 5 showed the effect of cultivars on pod nutri-tional value of beans expressed as the content of NPK, Faa,

Table 3 Effect of cultivars and brassinosteroids foliar application on yield and quality of bean plantsTreatments Pod weight

(g)Pod length (cm)

Total yield (kg m− 2)

2010 2011 Treat-ments

2011 2010 2011

Effect of BRsControl 2.36 2.24 10.70 10.67 0.72 0.7225 ppm 3.01 2.95 12.18 12.28 0.88 0.8750 ppm 4.56 4.64 14.30 14.60 0.86 0.85100 ppm 4.51 4.58 13.63 13.86 0.59 0.60LSD at 5 % 1.34 1.18 NS NS 0.08 0.13Effect of cultivarsPaulesta 3.57 3.56 12.34 12.46 0.70 0.70Oxzira 3.64 3.64 13.06 13.25 0.82 0.82LSD at 5 % NS NS NS NS 0.03 0.09Effect of the interactionControl Paulesta 2.28 2.16 10.25 10.18 0.55 0.56

Oxzira 2.44 2.33 11.15 11.16 0.89 0.8725 ppm Paulesta 2.43 2.32 11.55 11.60 0.88 0.87

Oxzira 3.58 3.57 12.80 12.96 0.88 0.8750 ppm Paulesta 4.89 5.00 13.50 13.73 0.81 0.81

Oxzira 4.23 4.28 15.10 15.47 0.91 0.89100 ppm Paulesta 4.69 4.78 14.05 14.32 0.57 0.58

Oxzira 4.33 4.38 13.20 13.40 0.61 0.62LSD at 5 % 2.28 2.16 10.25 10.18 0.55 0.56

Table 4 Effect of cultivars and brassinosteroids foliar application on N, P and K in bean podsTreatments N (%) P (%) K (%)

2010 2011 2010 2011 2010 2011Effect of BRsControl 3.16 3.14 0.38 0.41 1.58 1.5625 ppm 3.44 3.38 0.34 0.38 2.14 2.0350 ppm 3.40 3.34 0.49 0.52 2.60 2.50100 ppm 2.82 2.86 0.40 0.42 1.53 1.52LSD at 5 % 0.17 0.13 0.03 0.05 0.16 0.22Effect of cultivarsPaulesta 3.04 3.04 0.38 0.41 2.10 2.00Oxzira 3.37 3.32 0.43 0.46 1.82 1.80LSD at 5 % 0.22 0.15 NS NS NS NSEffect of the interactionControl Paulesta 3.25 3.22 0.44 0.47 1.59 1.57

Oxzira 3.06 3.06 0.32 0.36 1.57 1.5525 ppm Paulesta 3.25 3.22 0.36 0.39 2.68 2.49

Oxzira 3.63 3.54 0.33 0.37 1.59 1.5750 ppm Paulesta 2.96 2.98 0.31 0.36 2.58 2.40

Oxzira 3.83 3.70 0.67 0.69 2.62 2.60100 ppm Paulesta 2.68 2.74 0.41 0.42 1.57 1.55

Oxzira 2.96 2.98 0.39 0.42 1.49 1.49LSD at 5 % 0.21 0.17 0.06 0.09 0.10 0.15

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protein and phenolic contents in pods. ‘Oxira’ cultivar sig-nificantly had higher nitrogen in pods and pod protein con-tent in the two seasons. Whereas, phosphorus in pods and phenolic content were insignificantly affected from cultivar. On the contrary, the ‘Paulesta’ cultivar recorded higher con-tent of K and Faa in pods. These results were true and simi-lar in both seasons.

Effect of the Interaction

interaction between cultivars and BRs foliar application had a significant effect on NPK (Table 4). The highest NPK (%) was recorded using the ‘Oxira’ cultivar sprayed with BRs of 50 ppm in the two tested seasons and the lowest N and P % using the ‘Paulesta’ cultivar sprayed with the same con-centrations. Moreover, low K % was recorded regarding the ‘Oxira’ cultivar sprayed with BRs of 100 ppm.

‘Oxira’ cultivar significantly provided the highest value of protein in pods, when sprayed with BRs of 25 and 50 ppm, respectively (Table 5). However, the ‘Paulesta’ cultivar showed the highest pod phenolic content and pod Faa, when sprayed with BRs of 25 and 100 ppm, respec-tively. The lowest phenolic content and Faa in pods were recorded using the ‘Oxira’ cultivar when sprayed with BRs of 100 ppm and control treatment, respectively.

Discussion

Effect of BRs

Brassinosteroids (BRs) are a group of naturally occurring plant steroids that are important for a broad spectrum of cellular and physiological processes (Xia et al. 2009). The occurrence of BRs has been demonstrated in almost every part of the plant, such as pollen, flower buds, fruits, seeds, vascular cambium, leaves, shoots and roots. BRs have key functions in a range of developmental processes (Bajguz and Hayat 2009). Firstly discovered and isolated by grove et al. (1979), they are recently used exogenously as a natu-ral product to treat plants, in order to enhance their physi-ological activity, stimulate growth, increase yield and/or overcome inconvenient and unfavorable environmental conditions.

The stimulating effect of BRs on vegetative growth char-acters has been reported by several investigators. Howell et al. (2007), and Kang and guo (2011), for instance, stated that BRs stimulate elongation, cell division and differen-tiation, all of which furthermore promote plant growth. an improvement of shoot growth for different plants was also reported by Hayat et al. (2001), Nomura et al. (1997), Fariduddin et al. (2008), ali et al. (2008) and Hasan et al. (2008). BRs foliar spraying significantly enhanced vegeta-tive growth parameters, compared to the respective control treatment (Table 2). Spraying snap bean plants with BRs of 25 and 50 ppm increased plant length, number of leaves and shoots per plants, number of branches per plants as well as fresh and dry weight of leaves, while no significant differ-ences were measured between the values 25 and 100 ppm. Piñol and Simon (2009) and Jian et al. (2012) reported that treatments with BRs cause an increase in the number of leaves of bean and cucumber, respectively. all these results are in agreement with the results of the here presented study. BRs foliar spraying significantly enhanced vegetative growth parameters of snap beans compared to the respective control group. The highest vegetative growth parameters were obtained with BRs of 25 and 50 ppm (Table 2).

The fact that growth parameters in BRs treated plants were better than in untreated ones, has to be seen as evi-dence of a protective function of BRs to the photosynthetic apparatus from oxidative stress, induced by high temper-atures. This is in agreement with the findings of Ogweno et al. (2008). The authors stated that tomato plants treated with exogenously BRs significantly alleviated high-tem-perature-induced inhibition of photosynthesis by increasing its carboxylation efficiency and ability to regenerate RuBP under high-temperature stress. Other findings from Xia et al. (2009) emphasize the enhanced NaDPH oxidase activ-ity involved in the BR-induced stress tolerance. Conversely, Behnamnia et al. (2009) showed, during another oxidative

Table 5 Effect of cultivars and BRs foliar application on N, P and K in snap bean podsTreatments Faa

in podPhenolic acid in pod

Protein % in pod

2010 2011 2010 2011 2010 2011Effect of BRsControl 28.84 28.65 15.78 16.07 19.73 19.6225 ppm 30.18 30.10 19.01 19.59 21.52 21.1350 ppm 28.19 27.93 13.07 13.11 21.22 20.88100 ppm 32.97 33.15 11.42 11.32 17.63 17.86LSD at 5 % 1.45 2.42 1.12 1.36 1.06 0.81Effect of cultivarsPaulesta 33.81 34.06 14.28 14.43 18.98 18.99Oxzira 26.28 25.85 15.36 15.62 21.07 20.75LSD at 5 % 4.98 4.44 NS NS 1.38 0.94Effect of the interactionControl Paulesta 37.80 38.41 13.19 13.25 20.32 20.13

Oxzira 19.89 18.89 18.38 18.90 19.13 19.1225 ppm Paulesta 31.81 31.88 21.01 21.77 20.32 20.13

Oxzira 28.55 28.32 17.02 17.42 22.72 22.1450 ppm Paulesta 25.96 25.51 11.39 11.28 18.53 18.62

Oxzira 30.41 30.36 14.75 14.94 23.92 23.14100 ppm Paulesta 39.68 40.46 11.52 11.43 16.73 17.11

Oxzira 26.27 25.84 11.32 11.20 18.53 18.62LSD at 5 % 3.38 3.60 7.94 7.52 1.31 1.06

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stress situation, such as drought stress, an increase in the activity of antioxidant enzymes and antioxidative compound content of tomato plants after a pretreatment with BRs.

The climate conditions are to be seen as stressful, incon-venient and unfavourable by the delayed summer sowing of snap beans in our study. Bean plants are relatively sensi-tive to high temperatures, which negatively affect not only growth, but due to photosynthates, also other development parameters and, finally, the realized yield. The results of ibrahim et al. (2012) revealed that an early sowing on 1st of March showed the highest significant values for plant growth and dry weight of Phaseolus vulgaris l., compared with late or “heat stress-summer” planting. Saini and Negi (1998) and (Karas et al. 1999) found an increase in vegeta-tive growth of Phaseolus vulgaris l. on early sowing dates. according to Masaya and White (1991) beans are adapted to relatively cool climatic conditions with optimal average daily temperatures of 25 °C for reproductive development.

However, due to a treatment with BRs these detrimen-tal environmental conditions effects could be mitigated. The results from the present study clearly demonstrate the effectiveness of BRs in the improvement of the overall fruit yield of snap beans. an increase in fruit yield per plant was observed in bean plants, especially in comparison to non-treated plants (Table 3). These results are in agreement with findings from other authors in different crops. For instance, Torres and Núñez (1997) reported that BRs applications have resulted in a yield increase for potatoes, whereas a beneficial effect of BRs foliar spraying on fruit yield was also observed under heat stress, which was mainly due to the stimulation of fruit set and an increase in fruit number (Singh and Shono 2005).

increases in snap bean yields from BR treatments directly correlat with the increase in pod weight, and length (Table 3) as well as other pod quality parameters. an exog-enous BRs application significantly enhanced NPK content in pods (Tables 4 and 5). The increase of N content could be attributed to the fact that BRs caused an increase of NO3− absorption (Mai et al. 1989) and an increase in nitrate reduc-tase activity (ali et al. 2006; Fariduddin et al. 2008). Such increase in N content consequently led to an increase in the total free amino acid and protein content in pods (Tables 4 and 5). The highest values were observed in a concentration of 25 ppm, although the highest FFa in pods was reached at a concentration of 100 ppm. However, the protein content in pods expressed both in percent (Table 5) and calculated for the realized yield (data not shown) was always higher in concentrations of 25 and 50 ppm. From a practical point of view it is recommended to spray with BRs at a concen-tration of 25 ppm, since the differences for most measured plant growth and development parameters between 25 and 50 ppm were not significantly different.

Recently, different mechanisms that explain the often documented effect of BRs on plant growth and development are discussed in the literature. according to Bajguz and Hayat (2009) BRs can act efficiently in plants as immune modulators. BRs regulat stress response as a result of a com-plex sequence of biochemical reactions such as activation or suppression of key enzymatic reactions, induction of protein synthesis, and the production of various chemical defence compounds.

Dhaubhadel et al. (1999) showed that Brassica napus and tomato seedlings grown in the presence of 24-epibrassi-nol were significantly more tolerant to lethal heat treatment than control seedlings grown in the absence of the com-pound, increasing the basic thermotolerance, and result-ing in a higher accumulation of the four major classes of heat-shock proteins, compared to untreated seedlings. These results were in agreement with the findings of Kulaeva et al. (1991) and Wilen et al. (1995) in studies with wheat leaf cells and bromegrass cell suspension culture, respectively (hereby referring to the accumulation of a set of heat shock proteins). Protein synthesis was maintained in BR-treated wheat leaves at 43 °C, at levels similar to those at 23 °C, whereas in untreated leaves it decreased 2.5-fold at 43 °C as compared to samples at the control temperature (Kulaeva et al. 1991). in another study, Dhaubhadel et al. (2002) sug-gested that exogenous BRs treatments limit the loss of some of the components of the translational apparatus during prolonged heat stress, and increase the level of expression of some of the components of the translational machinery during recovery, which correlate with a more rapid resump-tion of cellular protein synthesis following heat stress and a higher survival rate.

in conclusion, this study indicates that exogenously BRs mitigated the detrimental effect of oxidative stress of high temperatures.

Effect of Cultivars

‘Oxzira’ cultivar showed a better plant growth and the high-est vegetative growth parameters and dry matter compared to the ‘Paulesta’ cultivar. Furthermore, ‘Oxira’ cultivar showed high pod yield and high pod nutritional value as compared to the ‘Paulesta’ cultivar. These results could be explained as a result of genetic differences between the two tested cultivars.

Conclusion

in conclusion, this study indicated that exogenously BRs could mitigate the detrimental effect of oxidative stress of high temperatures on snap bean plant growth and develop-ment. Thus, BRs may be used, in order to induce thermo-

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tolerance of snap bean plants cultivated out of season in delayed summer conditions. For practical application, it is suggested to spray the snap bean plants at a concentration of 25 ppm.

References

ali B, Hayat S, Hasan Sa, ahmad a (2006) Effect of root applied 28-homobrassinolide on the performance of Lycopersicon ecul-entum l. Sci Hortic 110:267–273

ali B, Hasan Sa, Hayat S, Hayat Q, Yadav S, Fariduddin Q, ahmad a (2008) a role of BRs in the amelioration of aluminium stress through antioxidant system in mung bean (Vigna radiate l. Wilc-zek). Environ Exp Bot 62:153–159

amer aH, Sawan OM, Salman SR (2002) Water requirements of snap bean (Phaseolus vulgaris l.) as affected by sowing date under newly reclaimed soil at shark El-Owinat region. J agric Sci, Man-soura Univ 27:6097–6107

aOaC (1990) Official methods of analysis. 15th edn. association of Official analytical Chemists, Washington DC

Bajguz a, Hayat S (2009) Effects of brassinosteroids on the plant responses to envi-ronmental stresses. Plant Physiol Biochem 47:1–8

Behnamnia M, Kalantari KM, Rezanejad F (2009) Exogenous appli-cation of brassinos-teroid alleviates drought-induced oxidative stress in Lycopersicon esculentum l. general appl Plant Physiol 35(1–2):22–34

Clouse SD, Sasse JM (1998) Brassinosteroids: essential regulators of plant growth and development. annu Rev Plant Physiol Plant Mol Biol 49:427–451

Danial HD, george CM (1972) Peach seed dormancy in relation to endogenous inhibitors and applied growth substances. J am Soc Hort Sci 17:651

Dhaubhadel S, Chaudhary S, Dobinson KF, Krishna P (1999) Treat-ment of 24-pibrassinolide, a brassinosteroid, increases the basic thermotolerance of Brassica napus and tomato seedlings. Plant Mol Biol 40:333–342

Fariduddin Q, Hasan Sa, ali B, Hayat S, ahmad a (2008) Effect of modes of application of 28-homobrassinolide on mung bean. Turk J Biol 32:17–21

gomez Ka, gomez aa (1984) Statistical procedures for agriculture research, 2nd edn. Wiley-interscience, New York

grove MD, Spencer Fg, Rohwededer WK, Mandava NB, Worley JF, Warthen JD Jr, Steffens gl, Flippen-anderson Jl, Cook JC Jr (1979) Brassinolide, a plant growth promoting steroid isolated from Brassica napus pollen. Nature 281:216–217

Hasan Sa, Hayat S, ali B, ahmad a (2008) 28-homobrassinolide protects chickpea (Cicer arietinum) from cadmium toxicity by stimulating antioxidants. Environ Pollut 151:60–66

Hayat S, ahmad a, Hussain a, Mobin M (2001) growth of wheat seedlings raised from the grains treated with 28-homobrassino-lide. acta Physiol Plant 23:27–30

Howell WM, Keller gE 3rd, Kirkpatrick JD, Jenkins Rl (2007) Effects of the plant steroidal hormone, 24-epibrassinolide, on the mitotic index and growth of onion (Allium cepa) root tips. genet Mol Res 6:50–58

ibrahim SK, Shalaby MaF, El-Said Zaki M, abou-Sedera Fa, abd allah MSa (2012) alleviation of high temperature stress on snap bean (Phaseolus vulgaris l.) by benzyl adenine and putrescine. J appl Sci Res 8(1):192–199

Jian YP, Cheng F, Zhou YH, Xia XJ, Shi K, Yu JQ (2012) interactive effects of CO2 enrichment and brassinosteroid on CO2 assimila-tion and photosynthetic electron transport in Cucumis sativus. Environ Exp Bot 75:98–106

Kang YY, guo SR (2011) Role of brassinosteroids on horticultural crops. in: Hayat S, ahmad a (eds) Brassinosteroids: a class of plant hormone. Springer, Dordrecht, pp. 269–288

Karas aN, Singer SM, Sawan OM, abou Hadid aF (1999) Water con-sumption of bean plants (Phaseolus vulgaris l.) as affected by sowing dates. Egypt J Hort 26:19–34

Konsens i, Ofir M, Kigel J (1991) The effect of temperature on the pro-duction and abscission of flowers and pods in snap bean (Phaseo-lus vulgaris l.). ann Bot 67:391–399

Krishna P (2003) Brassinosteroid-mediated stress responses. J Plant growth Regul 22(4):279–289

Kulaeva ON, Burkhanova Ea, Fedina aB, Khokhlova Va, Bokebayeva ga, Vorbrodt HM, adam g (1991) Effect of brassinosteroids on protein synthesis and plant cell ultrastructure under stress condi-tions. in: Cutler Hg, Yokota T, adam g (eds) Brassinosteroids: chemistry, bioactivity and applications, aCS symposium, Ser. 474:141–155. american Chemical Society, Washington, DC

Mai YY, lin JM, Zeng Xl, Pan RJ (1989) Effect of homobrassinolide of the activity of nitrate reductase in rice seedlings. Plant Physiol Commun 2:50–52

Masaya P, White JW (1991) adaptation to photoperiod and tempera-ture. in: Van Schoonhoven a, Voysest O (eds) Common beans-research for crop improvement. CiaT, Cali, pp. 445–500

Mitchell JW, Mandava NB, Worley JF, Plimmer JR, Smith MV (1970) Brassins: a new family of plant hormones from rape pollen. Nature 225:1065–1066

Monterroso Va, Wien HC (1990) Flower and pod abscission due to heat stress in beans. J am Soc Hort Sci 115:631–634

Nomura T, Nakayama M, Reid JB, Takeuchi Y, Yokota T (1997) Blockage of brassinosteroid biosynthesis and sensitivity causes dwarfism in garden pea. Plant Physiol 113:31–37

Ogweno J, Xing Song, Kai Shi, Hu Wen, Wei Mao, Yan Zhou, Yu Jing, Salvador No-gués (2008) Brassinosteroids alleviate heat-induced inhibition of photosynthesis by increasing carboxylation effi-ciency and enhancing antioxidant systems in Lycopersicon escul-entum l. J Plant growth Regul 27:49–57

Omae H, Kumar a, Kashiwaba K, Shono M (2006) influence of high temperature on morphological characters, biomass allocation, and yield components in snap bean (Phaseolus vulgaris l.). Plant Product Sci 9(3):200–205

Pinol R, Simon E (2009) Effect of 24-epibrassinolide on chlorophyll fluorescence and photosynthetic CO2 assimilation in Vicia faba plants treated with the photosynthesis-inhibiting herbicide terbu-tryn. J Plant growth Regul 28:97–105

Plummer D (1978) an introduction to practical biochemistry, 2nd edn. Mcgraw-Hall Mook, london, pp 144

Prasad PVV, Boote KJ, allen lH Jr (2002) Effects of elevated tem-perature and carbon dioxide on seed-set and yield of kidney bean (Phaseolus vulgaris l.). global Change Biol 8:710–721

Saini JP, Negi SC (1998) Effect of sowing on growth and yield of french bean (Phaseolus vulgaris l.) under dry temperate condi-tion. indian J agron 43:110–113

Singh i, Shono M (2005) Physiological and molecular effects of 24-epibrassinolide, a brassinosteroid, on thermotolerance of tomato. Plant growth Regul 47:111–119

Torres W, Nunez M (1997) The application of biobras-6 and its effect on potato (Solanum tuberosum l.) yields. Cultivos Trop 18:8–10

Wilen RW, Sacco M, gusta lV Krishna P (1995) Effects of 24- epi-brassinolide on freezing and thermotolerance of bromgrass (Bro-mus inermis) cell cultures. Plant Physiol 95:195–202

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452453454455456457458459460461462463464465466467468469470471472473474475476477478479480481482483484485486487488489490491492493494495496497498499500501502503504505506507508509510511512513514515

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Xia XJ, Wang YJ, Zhou YH, Tao Y, Mao WH, Shi K, asami T, Chen Z, Yu JQ (2009) Reactive oxygen species are involved in brassi-nosteroid-induced stress tolerance in cucumber. Plant Physiol 150(2):801–814

Zhu BC, Su J, Cham MC, Verma DPS, Fan Yl, Wu R (1998) Overex-pression of pyrroline-5-carboxylate synthetase gene and analysis of tolerance to water stress and salt stress in transgenic rice. Plant Sci 139:41–48

Abdelmohsin Mahmoud El-Bassiony, Prof. of vegetable research, Vegetable Research Department, National Research Center, Dokki, Cairo, Egypt. His Fields of interest and expertise are: Organic agri-culture, vegetable production, protected cultivation, soilless culture, plant nutrition for vegetable crops, Mushroom production.

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Questions to the Author(s)

AQ1.  Please provide keywords in German and English. AQ2.  Please provide German title, German keywords, and English keywords for this manuscript. AQ3.  “Dhaubhadel et al. 2002” is cited in the text but is not given in the reference list. Please provide a full reference or delete the citation. AQ4.  The sentence “In conclusion, this study… high temperatures” appears in sections “Discussions” and “Conclusion”. Please check whether this sentence can be deleted from the “Discussions” section. AQ5.  “Amer et al. 2002” is not cited in the text. Please provide the citation or delete the entry from the reference list.