Zn`s effect on health and growth of Eruca Sativa
Research article: The effects
of Zinc soil contamination on
the health and growth of rocket
Eruca Sativa (Mill)
Author: Ryan Pullen
Supervisor: Nigel Chaffey
Institution: Bath Spa University
Year: 2013/14
Key words: Zinc, Eruca Sativa, Heavy metals, Chlorosis, growth, Health
Abstract length: 190 words, 1101 characters (inc spaces)
Word count: 4998 words, 31731 characters (inc spaces)
Including critique and future work: 5792 words, 36743 characters
(inc spaces)
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Zn`s effect on health and growth of Eruca Sativa
Zn`s effect on health and growth of Eruca Sativa
Abstract
2
Zn`s effect on health and growth of Eruca Sativa
Background: Rocket (Eruca Sativa) is grown crop around the world and is
commonly used in salad; it is a fairly tolerant species to drought
and poor quality soils. Zn is an essential micronutrient for plants
and plays roles in many different physiological functions, however
due to the high levels of pollution soils have become contaminated.
Phyto-extraction is a method of using higher plants to translocate
organic contaminants into harvestable biomass to remediate soils.
Methodology: Eruca Sativa was grown in 3 treatments and a control over
the course of 49 days over which time observations on health and
growth were made, they were then harvested and the fresh weight, dry
weights, Zn uptake in leaf and shoot, Chlorophyll content were
measured.
Results/discussion: Significantly reduced biomass production at
highest treatment (1200mg/kg) suggesting toxic contamination,
significantly greater chlorophyll production at low treatment than
at higher treatment suggesting Zn effects chlorophyll content but
can become toxic and cause chlorosis. Significant Zn uptake as
treatments got higher, At 1200mg/kg median Zn content was
4857.14mg/L at highest treatment uptake boomed suggesting that Eruca
Sativa is a tolerant plant which excludes contaminants until a
threshold is reached, between 800 and 1200 mg/kg contamination.
3
Zn`s effect on health and growth of Eruca Sativa
Abstract
Introduction Eruca sativa - 6 Heavy metals - 6 Plants relationship with heavy metals - 7 Bioavailability - 8 Zinc - 8 Heavy metal tolerance - 9 Remediation techniques - 10 Phyto extraction - 11 Aims - 12 Objectives - 12
Methodology
Greenhouse
Greenhouse conditions - 13 Sowing and potting - 13 Sample size and treatment - 13 Randomizing - 14 Observations and monitoring - 14 Harvesting - 14
Laboratory analysis
Fresh and dry weights - 15 Atomic absorption spectrophotometry (AAS) - 15 Soil tests - 15 Chlorophyll content - 16 Statistical analysis - 16
Results
Fresh weight - 17 Dry weight - 18 Zn uptake in leaf - 19
4
Zn`s effect on health and growth of Eruca Sativa
Zn uptake in shoot - 20 Total Zn uptake - 21 Leaf vs. shoot comparison - 22 Chlorophyll content - 23 Chlorophyll ratio - 26 Purpling - 26 Flowering - 27
Discussion
Zn`s physiological roles - 28 Zn toxicity - 30 Eruca sativa tolerance - 30 eruca sativa phyto extraction potential - 32 conclusion - 34
Critique - 35
Future work - 36
Acknowledgments - 38
References - 39
Appendix - 53
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Zn`s effect on health and growth of Eruca Sativa
Eruca sativa
Rocket (Eruca Sativa) is a member of the Brassicacceae family (Abassi et
al., 2013).and is an endemic species originating from the
Mediterranean (Barlas et al., 2011; Abassi et al., 2013) and is now
grown worldwide due to its tolerance of dry, disturbed land (Al-
quarainy, 2009)
It grows from 20 to 50cm in height (Barlas et al., 2011) Eruca Sativa
growth is quicker in Higher temperatures and longer day lengths
(Dolezalova et al., 2013). Eruca Sativa is known to bolt into flowers in
longer days (Jaske, 2012) although it is usually harvested when the
leaves are young for salads (Dolezalova et al., 2013) Eruca Sativa has
also got medicinal purposes (Abassi et al., 2013) and is known to
6
Zn`s effect on health and growth of Eruca Sativa
have many properties such as astringent, digestive, emollient,
laxative, tonic, stomachic, anti-inflammatory for colitis and
stimulant properties (Barlas et al., 2011; Abassi et al., 2013). Eruca
Sativa bio-components has also got cancer prevention potential
(Michael et al.,2011; Ambrosone and Tang, 2009; Higdon et al., 2007) as
well as use as an aphrodisiac (Dolezalova et al., 2013).
It is an easily grown species (Barlas et al., 2011) and has natural
resistance to drought and pathogenic invasion (Abassi et al., 2013).
Eruca Sativa has been noted as a highly tolerant species and can
accumulate some HM (Saleh, 2001), it is notably tolerant of salts
(Ashraf & Harris, 2004)
Heavy metals
“Heavy Metals” (HM) are defined as a metallic element with a
density between 4-5g/cm3 (Jarup, 2003; Nagajyoti et al., 2010).
Commonly found HM include Lead (Pb), chromium (Cr), arsenic (As),
zinc (Zn), cadmium (Cd), copper (Cu) (Wuana & Okieimen, 2011). In
toxic concentrations HMs can cause damage to the ecology,
environmental, nutritional and evolutionary characteristics of the
polluted area (Babula et al., 2008). HM pollution boomed during the
1900`s industrial revolution leading to high levels of soil, water
and atmospheric contamination (Jarup, 2003).
7
Zn`s effect on health and growth of Eruca Sativa
Soils are HMs sinks (Wuana & Okieimen, 2011) HMs originate from the
earths crustal rock and are released through weathering processes
(Wuana & Okieimen, 2011) although its anthropogenic sources such as;
fertilisers, mine tailings, pesticides, sewage sludge and smelting
have caused unnaturally high contamination (Wuana & Okieimen, 2011).
Concentration of HMs persists as they are not degraded from
microbial or chemicals like organics, reducing soil quality (Lasat,
2002). Prolonged contamination of soils severely reduces the soil
quality (Oliviera and Pampulha, 2006) However changes to chemical
form are possible (Wuana & Okieimen, 2011). Humans require HMs as
they are essential micro-nutrients but whilst essential, HMs can be
poisonous at toxic levels (Duiribe et al., 2007). There is currently
a widespread issue with dietary Zn deficiency (Myers, 2014).
HMs and plants
HMs and plants have complex relationships. HMs is essential
nutrients in trace concentrations for healthy growth as Plants
require the nutrients for essential physiological functions (Tangahu
et al., 2011). Deficient or toxic concentrations can cause
disruptions to essential functions leading to poor health or death.
(Nagajyoti et al., 2010) The degree of toxicity or deficiency the
plant has to tolerate to survive is affected by Metal Form and
8
Zn`s effect on health and growth of Eruca Sativa
concentration, bioavailability and species (Nagajyoti et al., 2010).
High and low concentrations of HM in soil can negatively affect crop
growth, as these metals interfere with metabolic functions in
plants, including inhibition of photosynthesis and respiration and
degeneration of main cell organelles, even leading to death of
plants (2001; Schmidt, 2003; Schwartz et al., 2003).
Bioavailability
Bioavailability is the measure Zn available for uptake,
Bioavailability of Zinc primarily depends on the concentration in
the soil, Ph of the soil and its chemical form (Lin & Aarts 2012;
Pilon-Smits, 2005) exposure time, species, environmental conditions
which also affects bioavailability (Lin & Aarts 2012). Zn is more
available in low Ph soil as Ph reduces Zn concentration in solution
( Rout & Das, 2003). Zn interacts with other HM in the soil. Zn
balances with other HMs affects the bioavailability. An essential
balance for Zn is P, high levels of P cause Zn deficiencies
(Mousavi, 2011) the relationship between Zn vs. Cu. Is such that if
one is taken up in high concentration it inhibits the uptake of the
other (Mousavi, 2011) whilst Zn vs. Fe have a sympathetic
9
Zn`s effect on health and growth of Eruca Sativa
relationship, however Zn in excess can severely reduce Fe uptake
(Mousavi et al., 2013).
Zinc
Zn is an essential micro-nutrient for physiological functions. Zn is
a building block for enzymes; in addition many enzymatic reactions
are activated by zinc (Mousavi, 2011). Zinc exerts a great
influence on many plant life processes, such as; Nitrogen metabolism
and uptake of nitrogen and protein quality; photosynthesis and
chlorophyll synthesis, carbon anhydrase activity; resistance to
abiotic and biotic stresses and protection against oxidative damage
(Cakmak, 2000; Shulin et al., 2009).
Figure 1: Zinc concentration and its effect on the likelihood
of the symptoms deficiency and toxicity being present (Lin &
Aarts, 2012).
Zn deficiency is commonly reported in crop growth (Assuncao et al.,
2010) Zn deficient plants suffer from physiological stress caused by
10
Zn`s effect on health and growth of Eruca Sativa
enzyme dysfunction and other metabolic function disruptions
(Mousavi, 2011) Symptoms of deficiency include stunted growth,
inter-venial chlorosis in younger leaves, necrotic tips and
photosynthetic problems (Rout & das, 2003).
Zn toxicity leads plants to suffer from physiological stress caused
by enzyme dysfunction and other metabolic function disruptions
(Mousavi, 2011). Zn excess can cause: ATP synthesis (Mousavi et
al.,2013), other symptoms include chlorosis, smaller leaves and
necrotic leaf tips (Rout & Das, 2003; Sagardoy et al.,2008)
HM tolerance
HM tolerant species fit into three strategic types HM tolerance;
excluders, indicators and accumulators (Ghosh & Singh 2005; Mganga,
2011; Bert et al.,2000)
Figure 2 – Strategies for coping with high HM contamination in
soil (Ghosh & Singh 2005)
11
Zn`s effect on health and growth of Eruca Sativa
Indicator species accumulate metals into tissues to a level that
reflects the concentration in the soil regardless of high or low
levels (Bakker, 2008). Excluders prevent metals from entering the
above ground tissues however there`s still accumulation in the roots
(Ghosh & Singh 2005; Dahmani- Muller et al., 2000 ), HM ions are
excluded by cell wall binding and many other mechanisms
increasing tolerance (Hall, 2002).
Accumulator species concentrate the HMs in above ground tissues;
these are species known to accumulate metals at concentrations
higher than in the soil (Maestri et al., 2010, Babula et al., 2008;).
To be classified an accumulator HM uptake must exceed 10 000 mg/kg
Zn and Mn, 1000 mg/kg for Co, Cu, Ni, As and Se, and 100 mg/kg Cd
(Mcgrath and Zhou, 2002) there are between 400 and 450 known hyper
accumulators (Padamavathiamma & Lu, 2007).
Bio remediation
Bioremediation are techniques for reducing HM contamination, many
techniques like chemical washing and physical removal are available
(Sarma, 2011; Garbisu & alkorta, 2001; Lone et al., 2008) However
such methods cause irreversible changes to the soil, reducing its
fertility (Padamavathiamma & Lu, 2007; Diacono & Montemurra, 2009).
Another method used is phytoremediation; the use of HM tolerant
plants to translocate or alter contaminants to reduce their
concentrations (Pal et al., 2010) .
12
Zn`s effect on health and growth of Eruca Sativa
(Table 1) – Commonly described methods of phytoremediation adapted
from, (Ghosh, Padamavathiamma & Lu, 2007; Vamerali et al., 2009)
process mechanism contaminant
Phyto-extraction Hyper-accumulation of
metals in harvestable
tissues
Inorganics
Phyto-stabilisation Metal retention in the
root
Inorganics
Phyto-volatilization Evaporation of
dissolved contaminants
from leaves
Organics/Inorganics
Phyto-transormation/
Phyto-degradation
Microbial
degradation/speciation
of contaminant in
plant tissues
Organics/inorganics
Phyto-filtration/
rhizo-filtration
Root and Rhizome
accumulation
Organics/Inorgancs
Phyto-extraction
Phyto-extraction is the remediation technique which uses higher
plants to concentrate HM in harvestable tissues (Pal et al., 2010).
Hyper-accumulators take up and translocate HMs from the soil into
harvestable tissues (Hooda, 2007). Metal chelation agents can be
13
Zn`s effect on health and growth of Eruca Sativa
used for encouraging the uptake of HM (Vangrosvield et al., 2009;
Tandy et al., 2004;Khan et al., 2000).
Effective species for Phyto-extraction must fit these criteria
(Padamavathiamma & Lu 2007)
Large Biomass
High accumulation of metals
Has the ability to transport large amounts of metals into
harvestable biomass
Genetics for high tolerance to metals
Rapid growth
Resistant to disease and pests.
The harvested waste material has many applications; combustion for
energy, metal recycling or production of fertilisers (Alkorta et
al., 2004)
There is a desire for effective, cheap and green techniques (Pilon-
smits, 2005). Phytoextraction is currently receiving a lot of
attention as it is both cheap and green (Badr et al., 2012). However
The disadvantage of phytoremediation is the time it takes, time
taken for a species to grow is significantly slower than chemical or
physical approaches (Vangrosvield et al., 2009) and should be
considered a long term solution (Prasad & frietas,2003)
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Zn`s effect on health and growth of Eruca Sativa
Aims:
Zn is an essential element for plants and can encourage great health
at optimised treatments but can become deficient or toxic and cause
ill health. Phyto-extraction is an exciting research topic looking
into the abilities of plant species of accumulating HMs in very high
concentrations. The aim of this study is to evaluate the effects of
Zn on the health and growth and the tolerance and accumulation of Zn
in Eruca Sativa.
Objectives:
To grow Eruca Sativa in varied Zn concentrations
To assess the growth and health of the plants, comparing
chlorophyll concentration, Zn accumulation and biomass
production
To observe several health indicators such as chlorosis and or
purpling, shoot size, death and flowering.
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Zn`s effect on health and growth of Eruca Sativa
Methodology
Greenhouse Maintenance
The greenhouse was kept at consistent temperature and light levels
by heaters and growth lights extending hours of light to 16 and
ensuring that the temperature does not go below 0oC.
Sewing and potting
300 seeds were sown in seed trays in the compost with vermiculite
and perlite for improved germination and seedling growth conditions
(takahashi et al., 2006;Sari &Karaipekil, 2007). Once germinated 3
individuals were transferred into 1L pots, and then grown for 7 days
so any that had died or failed to make the transfer unharmed were
replaced.
16
Zn`s effect on health and growth of Eruca Sativa
Sample size and Treatment application
The treatments selected were 400mg/kg, 800mg/kg and 1200mg/kg of
zinc adapted from a similar study by Ozdener & Aydin (2010), these
will hereafter be referred to control, low, medium, high treatments.
These were made up by diluting 219.78g of ZnSO4.7H2O (Zinc Sulphate)
into 500Ml for 100mg/kg and applying appropriate Ml for each
treatment via a pipette. To calculate the volume of compound to be
administered for each treatment this formula was used.
Compound
weight (g) =
mg/kg element X
dilution rate X 100
1000
%
element
(full Calculations in appendices A).
For each treatment there were 20 pots with 3 individuals in
totalling 60 plants per treatment (total = 240). Half the pots were
designated for study A (fresh, and dry weights and Zn uptake )and B
(chlorophyll content)(Figure 3). Each was labelled and the growth
phase had begun.
17
Zn`s effect on health and growth of Eruca Sativa
Figure 3: distribution of the samples by treatment and which study
there were applicable to.
Randomising
The pots were randomly assigned its growing position in the
greenhouse space using a random number generator (appendices B)
in excel (Microsoft, 2010).
Observations and monitoring
After the initial 21 days of the growth phase weekly observations
began. Observations of leaves exhibiting purpling and how dark the
purpling had become, quantity dying or dead plants, shoot length,
size of leaf, and from week 5 onwards individuals that had flower
and the number of flowers.
Harvesting
The harvesting process consisted of cutting the shoot at the base
just above the soil, hence cutting the above ground portion from the
18
Zn`s effect on health and growth of Eruca Sativa
roots, this was done delicately to get the whole shoot rather than
cut it short. The leaf and shoot tissues from each pot were then
carefully placed in labelled paper bags.
Laboratory analysis
Fresh and dry weights
Fresh weights were taken immediately after harvesting. The samples
were then left over night in an oven to dry them out. The next day
the samples were checked to ensure they were completely dried, once
fully dried dry weights were taken.
Atomic absorption spectrophotometer (AAS)
The leaf tissue was separated from the shoot and then approximately
1g was weighed out for leaf and another 1g for shoot, However for
the samples from the 1200 treatment not enough material for each
compartment therefore plant materials mixed. The exact weight was
recorded ( 2 d.m).
Once weighed samples were digested in labelled kjeldatherm tubes in
the fume cupboard with 5Ml nitric acid. Samples were then refluxed
at 120ᵒC for 20 minutes followed by 20 minutes at 160ᵒC. once cooled
the digested solution was filtered through whatman no.54 paper into
50Ml volumetric flasks made up with distilled water.
19
Zn`s effect on health and growth of Eruca Sativa
Zn concentration was determined using the AAS which had been
calibrated. Throughout testing many of the samples were read as
“OVER” so a 1 in 10 dilution was required for each sample. (RSC,
undated).
Soil tests
The compost used for the growth phase was tested for its Ph – using
the method described in Eckert and Sims (2011) and organic matter –
using the method described in Mathiessen (2006).
Chlorophyll content
Chlorophyll content was determined by using the method set out in
Inskeep & Bloom (1985)
0.1g of leaf was weighed out and then macerated in 12.5Ml of acetone
twice. The liquid was decanted into a separate clearly labelled
tube. This solution was equally weighed out between two centrifuge
tubes and spun in the centrifuge at 3000rpm for 3 minutes. Once
completed the solution was poured into a 50Ml volumetric flask. The
solution was then decanted into cuvettes.
20
Zn`s effect on health and growth of Eruca Sativa
In the spectrophotometer the absorbance of the samples at
wavelengths 646.6 and 663.6 were recorded 4 samples at a time.
Chlorophyll content calculated from equations in Porro (2002).
Chlorophyll a (µg/Ml) = 12.25 (A663.6) – 2.55
(A646.6)
Chlorophyll b (µg/Ml) = 20.31 (A646.6) – 4.91
(A663.6)
Total chl (µg/Ml) = 17.76 (A646.6) + 7.34 (A663.6)
Statistical analysis
Data was assessed for normality using the Sharpiro-Wilk test prior
to running further statistical analysis, the results of which led to
the following: data with more than two sets, For non-parametric
data Kruskal Wallis was used, for parametric (ANOVA) with Tukeys HSD
test to determine where the significance was. For data with only 2
sets, 2 sample T tests were used.
21
Zn`s effect on health and growth of Eruca Sativa
Results
Fresh weights
trace 400 800 12000
2
4
6
8
10
12
14
Applied treatment Zn (ppm)
Fresh weight (g)
Figure 4: Mean Fresh weight post-harvest per applied treatment
of Zn, with 95% confidence error bars.
Figure 4 shows There was significantly less fresh weight from those
in the high treatment (Mean 5.8g; Median: 5.89g) than each other
treatment (p value = 0.007) but no significance between the medium
treatment (Mean: 12.56g), low treatment (Mean 10.3g) and control
(Mean: 9.51).
22
Zn`s effect on health and growth of Eruca Sativa
Dry weights
trace 400 800 12000.00
0.50
1.00
1.50
2.00
2.50
Applied treatment zn (ppm)
Dry weight (g)
Figure 5: Mean dry weight post-harvest per applied treatment
mg/kg of Zn, with 95% confidence error bars. 800> 400> control
> 1200
Data in figure 5 shows Significantly less dry weight from those in
the high treatment (Mean, Median) than that of each of the other
23
Zn`s effect on health and growth of Eruca Sativa
treatments (p value = 0.002) but no significantly different Mean dry
weights between the rest.
Tissue Zn content
trace 400 8000
100
200
300
400
500
600
700
800
Applied Zn treatment (ppm)
Zn concentration in tissue (mg/L)
Figure 6: Mean Zn content (mg/L) in the shoot tissues from each
treatment, with 95% confidence intervals.
24
Zn`s effect on health and growth of Eruca Sativa
Significantly different shoot Zn uptake between each treatment
(kruskal wallis = 20.66, P value <0.001) medium treatment (Median:
512.82 mg/L) had significantly greater uptake than low treatment
(Median: 166.67 mg/L) which was significantly greater than the
control (Median: 27.86 mg/L). (Figure 6)
trace 400 8000
100
200
300
400
500
600
700
800
900
1000
Applied Zn treatment (ppm)
Zn concentration in tissue (mg/L)
25
Zn`s effect on health and growth of Eruca Sativa
Figure 7: Mean Zn content (mg/L) in leaf tissue from each
treatment, with 95% confidence intervals.
Significantly different leaf Zn uptake between each treatment
(kruskal wallis = 18.28, P value <0.001) medium treatment (Median:
758.62 mg/L) had significantly greater uptake than low treatment
(Median: 464.65 mg/L) which had significance over control (Median:
24.83 mg/L). (Figure 7)
trace 400 800 12000
500100015002000250030003500400045005000
Applied Zn treatment (ppm)
Zn concentration in ttissue (mg/L)
0 400 800 12000
1000
2000
3000
4000
5000
6000
Applied Zn treatment (ppm)
Zn concentratin in tissues
Figure 8: Mean Zn content (mg/L) in the all harvestable tissues
(leaf+Shoot) from each treatment, with 95% confidence
intervals. Figure 9: Individual plot of total Zn uptake,
displaying the increasing Zn uptake throughout the treatments.
26
Zn`s effect on health and growth of Eruca Sativa
Figures 8,9 illustrate the Significantly different total Zn uptake
between each treatment (kruskal wallis = 31.55, P value <0.001) the
Highest treatment had significantly greater Zn uptake (Median:
4857.14mg/L) medium treatment (Median: 351.33 mg/L) had
significantly greater uptake than low treatment (Median: 193.33
mg/L) which had further significance over the control (Median: 15.44
mg/L).
Leaf vs. shoot comparison
27
Zn`s effect on health and growth of Eruca Sativa
trace 400 8000
100
200
300
400
500
600
700
800
900
1000
leafshoot
Applied Zn treatment (ppm)
Zn concentration in tissue (mg/L)
Figure 10: Zn content comparisons between leaf and shoot
material for each treatment, showing the translocating ability
of eruca sativa.
There was significantly greater Zn content in the leaves than in the
shoot in the low treatment. Leaf content (Mean: 532mg/L) was
significantly greater than the Zn content in the shoots (221 mg/L).
(P value = 0.001). However there was No significance at medium
treatment Leaf Tissue (Mean: 945 mg/L) was not significantly greater
than shoot tissues (Mean: 752mg/L) (P value = 0.462). Neither was
there significance for the control. Shoot Tissue (Mean: 36.4 mg/L)
28
Zn`s effect on health and growth of Eruca Sativa
was not significantly greater than leaf tissues (Mean: 31.7 mg/L).
(P value = 0.605). (Figure 10)
Chlorophyll A content
trace 400 800 12000
0.5
1
1.5
2
2.5
3
3.5
Applied treatment Zn (ppm)
chlorophyll content (µg/ml)
Figure 11: Chlorophyll A content in leaf tissues at each
treatment, using Porro`s (2002) chlorophyll content equation.
Low > medium > high > control.
29
Zn`s effect on health and growth of Eruca Sativa
In figure 11 its shown that Chlorophyll A content in the low
treatment (Mean: 2.91ug/Ml, Median:2.96ug/Ml) was significantly
greater than each of the other treatments. Each of the other
treatments were not significantly different to each other. The
Means of each were almost level 1.63 ug/Ml, 1.68 ug/Ml, 1.65 ug/Ml
for control, 800 and 1200 treatments respectively.
Chlorophyll B Content
30
Zn`s effect on health and growth of Eruca Sativa
trace 400 800 12000
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
Applied treatment Zn (ppm)
Chlorophyll content (ug/ml)
Figure 12: Chlorophyll A content in leaf tissues at each
treatment, using porro`s (2002) chlorophyll content equations
Low>medium>control>high.
Chlorophyll B content in the low treatment (Mean: 2.91ug/Ml,
Median:2.96ug/Ml) was significantly greater than the control and the
high treatments but not significantly greater than the medium
treatment (Mean: 0.62ug/Ml, Median: 0.64ug/Ml). (P value = 0.009)
(figure 12).
31
Zn`s effect on health and growth of Eruca Sativa
Figure 13: total Chlorophyll content in leaf tissues at each
treatment, using porro`s (2002) chlorophyll content equation.
Low> medium> control> high.
Mean total chlorophyll content of those grown in the low treatment
(Mean: 3.79ug/Ml, Median: 3.74ug/Ml) treatment was significantly
greater (p value = 0.002) than the other treatments. There was no
significance between the other treatments; Means were 2.22 ug/Ml,
2.30 ug/Ml, 2.16 ug/Ml for control, medium and high treatments
respectively. (figure 13).
33
trace 400 800 12000
0.5
1
1.5
2
2.5
3
3.5
4
Appied treatment Zn (ppm)
Chlorophyll content (ug/ml)
Zn`s effect on health and growth of Eruca Sativa
Table 2: Leaf chlorophyll content ratio`s A/B in each treatments.
Chlorophyll ratio
A/B
contr
ol 400 800 1200
Mean
2.737
499
3.377
527
2.727
979
3.207
932
Media
n
2.792
959
3.449
15
2.578
887
3.304
642
No significantly different chlorophyll ratios were found, Zn
contamination had no effect on the ratios between A/B (P value =
0.423)
Purpling of leaves and mortality
Table 3: percentage of plants that are healthy or that are showing
necrosis, purpling on a weekly basis throughout the growth period.
34
Zn`s effect on health and growth of Eruca Sativa
Week
contro
l
400mg/
kg
800mg/
kg 1200mg/kg
necro
sis
purpli
ng
healt
hy
necro
sis
purpli
ng
health
y
necro
sis
purpli
ng
healt
hy
necros
is %
purpli
ng %
healt
hy
% %
leave
s % % %
Leaves
% % %
leave
s % % %
leave
s %
3 6.7 21.7 71.7 3.3 23.3 73.3 11.7 36.7 51.7 30.0 35.0 35.0
4 6.7 31.7 61.7 5.0 31.7 63.3 16.7 48.3 35.0 36.7 40.0 23.3
5 8.3 50.0 41.7 8.3 43.3 48.3 21.7 51.7 26.7 48.3 43.3 8.3
6 10.0 70.0 20.0 10.0 68.3 21.7 30.0 56.7 13.3 55.0 45.0 0.0
7 10.0 83.3 6.7 10.0 76.7 13.3 33.3 53.3 13.3 61.7 38.3 0.0
Flowering
35
Zn`s effect on health and growth of Eruca Sativa
trace 400 800 12000
10
20
30
40
50
60
70
80
90
100
% flowered% leafy
Figure 15: % of individuals which flowered for each treatment.
94.4% of the control treatment plants flowered, of the plants
grown in the low treatment 59.3% of the plants flowered. The
Medium treatment flowered 39.2% of the time and at the highest
treatment 40% of the plants flowered.
Discussion
36
Zn`s effect on health and growth of Eruca Sativa
Physiological roles of zinc
Zn is an essential nutrient for Chlorophyll biosynthesis which
requires Zn dependant enzymes throughout production (Mousavi, 2011),
Zn in excess reduces NADPH production in chloroplasts (Mousavi et
al.,2013), Zn can also effect photosynthesis efficiency (Cakmak,
2008) and the uptake of Fe, an essential nutrient for chlorophyll
biosynthesis (Broadly, 2007). In this study chlorophyll in plants
grown in low levels of Zn contamination were significantly greater
than the other treatments (figure 13) this is likely to be due to
Zn`s role in the biosynthesis of chlorophyll, increasing
photosynthetic ability and the fact that there was not Fe deficiency
as Zn concentrations were not excessive so chlorophyll production
was not inhibited.
There was a large reduction in chlorophyll content in the leaves of
those grown in the highest treatment (figures 11-13), this suggests
that Zn began to inhibit the production mechanisms of chlorophyll
became toxic causing the significant reductions in chlorophyll
content. at toxic levels Zn can cause many physiological disruptions
and a reduction in enzyme productivity (Mousavi, 2011)This suggests
that at between 800 and 1200 mg/kg symptoms of Zn toxicity have
begun to show, this was also an observation made in Ozdener & Aydin
(2010). Reduced chlorophyll could also be a resultant of Fe
deficiency brought on by the High levels of zinc (Mousavi, 2011).
37
Zn`s effect on health and growth of Eruca Sativa
Zn can contribute to the metabolism of N, an essential micro-
nutrient in the physiological mechanisms for chlorophyll pigments
biosynthesis (Mousavi, 2011). Neeto et al(2005) found that N
metabolism and bioavailability plays a major role in the ratio of
chlorophyll A and B that is synthesised. This study found that there
was no significance between the 2 pigments suggesting that Zn had no
effect on the pigment ratio. (Table 2)
Biomass production and plant growth requires many mechanisms, many
of these mechanisms require Zn (Mousavi et al., 2013) and these
include Auxin production a growth hormone which encourages the rapid
growth of plants (Zamimalova, 2003), cell cycles and cell elongation
which accelerates growth (Stals and Inze, 2001), This is paired with
Auxin`s role in cell cycle encouraging rapid mitosis (Beemster,
2003). Photosynthesis (Cakmak, 2008), Photosynthesis is essential
for maximising biomass production, poor photosynthetic ability can
lead to stunted growth (allen and Forsberg, 2001). Despite Zn
contributing to many physiological mechanisms associated with
biomass production there was no significant biomass production,
fresh and dry weights did slightly increase but not significantly
(figure 4) and there were no obvious signs of improvement with Zn
addition or Zn deficiency in the control, this again was also found
by Ozdener & Aydin (2010). Whilst it is likely that Zn does
encourage the growth of biomass this study does not provide solid
38
Zn`s effect on health and growth of Eruca Sativa
backing to the idea that Zn is highly essential for biomass
production.
However there were significant results from the Fresh and dry
weights for the high treatment, there was a significant drop in the
biomass production (fugures 4,5) A likely explanation for this is
that Zn was applied in toxic concentrations leading to the
inhibition and disruption to the physiological mechanism behind
biomass production. It is also possible that the excess of Zn has
also largely affected the Bioavailability of other nutrients in the
soil that are essential for biomass production, implying that
stunted growth and lack of biomass are symptoms of Zn toxicity,
concurrent with current beliefs (Rout & das, 2003; Mousavi et al.,
2013; Cakmak, 2008)
There are many uncertainties with the results of fresh weight and
dry weights. Firstly Fresh and dry weights are not the only growth
factors that can measure the biomass production and do not provide
the whole story. Shoot length and leaf size can further explain the
role of Zn of plants however Eruca Sativa bolted throughout the latter
portion of the growth phase (figure 15) When flowered Eruca Sativa has
very low biomass and is a tall thin shoot with limited leaf material
and small delicate flowers (Barlas et al 2011) Eruca Sativa is known to
be quick bolting into flower in longer warmer days (Jaske, 2012).
As this study was done with 16 hour lighting in the greenhouse, It
39
Zn`s effect on health and growth of Eruca Sativa
is likely that the day lengths causing many individuals to flower
has majorly effected the reliability of fresh and dry weights. Zn is
known to play many roles in biomass production, and there were
slight however non-significant increases in biomass production for
low and medium treatments which could imply Zn does improve yield
but other factors have prevented significant results.
Zn toxicity
Results from this study suggest that in the highest treatment
1200mg/kg Zn toxicity became a serious issue. There was reduced
biomass (figure 4) and chlorophyll production (figure 13) and
throughout growth phase observations were made that those grown in
the highest treatment were showing multiple symptoms of Zn toxicity
(table 3) and multiple individuals could not tolerate the high
levels of Zn. It is widely known that high HM contamination in soil
can prevent proper development (Smical et al., 2008). One potential
resultant to Zn stress in Eruca Sativa was the prevalence of purpling
of leaves at higher treatments. There was a higher proportion of
individuals exhibiting signs of inter-vienal and leaf tip purpling
at the higher treatments. The correlation between the prevalence of
40
Zn`s effect on health and growth of Eruca Sativa
leaves purpling and the contamination levels suggest that Zn in high
doses can lead to leaves becoming a discoloured purple.
Eruca Sativa tolerance
Eruca sativa can be grown worldwide due to its tolerance of dry,
disturbed land (Al- quarainy, 2009) and has the ability to grow in a
range of soils and conditions (Cag et al., 2004). Eruca Sativa growing
healthily in this diversity of conditions suggests that it is a
fairly hardy, versatile and tolerant species and Eruca Sativa has also
been observed to not only tolerate but to accumulate some HMs in low
concentrations. (Saleh, 2001).
There was a significant rise in the level of Zn uptake (figures 8,
9) in harvestable tissues for every rise in Zn treatment, as there
was in Ozdener & Aydin (2010). The increases between the controls,
low and medium treatments were steady, however the difference noted
between the medium and high treatment was far more significant.
Excluder species are tolerant to heavy metal contamination in the
growth medium however there is a threshold at which uptake rapidly
increases (Hall, 2002), The results from this and Ozdener & Aydin
(2010) suggest that Eruca Sativa has a Zn threshold at which uptake
increases and beings to suffer symptoms of toxicity (figure 8)
41
Zn`s effect on health and growth of Eruca Sativa
HM tolerant species fit into three strategic types HM tolerance;
excluders, indicators and accumulators (Ghosh & singh, 2005). The
patterns of HM uptake are distinctive and hence easy to identify
tolerance strategy deployed by plants under HM stress.
Results from this study (figure 9) suggest that Eruca Sativa employs an
exclusion strategy to tolerate the high levels of Zn contamination
in the higher treatments. Tolerance of HMs requires genetic
homeostatic ability to cope with the stress of highly contaminated
soils (Cobbett & Goldsburgh, 2002) exclusion mechanisms include
aspects such as transport uptake, distribution, chelation and/or
sequestration of metals into specific tissues (Grotz, 2006). Typical
exclusion specific mechanisms include binding of the cell wall,
sequestering ions in the cell vacuoles, complexion of the metal,
Heat shock protiens, organic and amino acids (Hall, 2002; Cobbett,
2000). The mechanisms allow Hm tolerance through exclusion, results
indicate that Eruca Sativa`s tolerance strategy is likely to be formed
by a combination of exclusion techniques. HM exclusion requires
plants genetics to provide adequate homeostatic functions, including
aspects such as transport uptake, distribution, chelation and/or
sequestration of metals into specific tissues (Grotz, 2006). This
studies results show that Eruca sativa is also tolerant to Zn
deficiency, as was the case in Ozdener & Aydin (2010). This
suggests that as a species it is capable of tolerating both slightly
42
Zn`s effect on health and growth of Eruca Sativa
toxic and deficient soils, suggesting that as a species it has a
highly adaptive and versatile range for regulation and tolerance.
Eruca sativa phyto-extraction potential
There are many factors that can account for a species capabilities
and efficiency as a hyper accumulator, Padamavathiamma and Lu (2007)
set out the following criteria for defining a species as a hyper
accumulator:
Large Biomass
As seen in this study Eruca Sativa only produces a few grams of biomass
(figure 4) and only grows up to 50cm with a small shoot it is a
small sized species much like fellow Brassicaccae (Barlas et al.,
2011) whereby the lack of biomass production reduces its suitability
(Tangahu et al.,2011; Pulford & Watson, 2003)
Total HM uptake = biomass x metal accumulation.
Eruca Sativa is therefore less suitable for phyto-extraction
High accumulation of metals
To be defined as a Zn hyper-accumulator a species must accumulate
10,000mg/kg of Zn (Mcgrath and Zhou, 2002). At the highest
contamination level that did not appear to be toxic, the medium
43
Zn`s effect on health and growth of Eruca Sativa
treatment Zn uptake was significantly less than this figure (figure
8) suggesting that Eruca Sativa is not a hyper accumulator of Zn. Zn
becomes far to toxic before accumulation reaches the level required
to be defined as a hyper-accumulator, limiting its extraction
qualities.
the ability to transport large amounts of metals into
harvestable biomass
the ability to uptake HMs from the soil and translocate them is an
essential ability for a hyper-accumulator, as is sequestering them
into organelles for use in physiological mechanisms and storage
(Williams et al., 2000; Tian et al., 2009)Results suggests that there
was limited transporting of Zn from the cell wall into the
harvestable plant, due to exclusion mechanisms deployed by Eruca
Sativa, there was also not any significance in the leaf and shoot
content suggesting that its ability to sequester HMs into organelles
is limited
Genetics for high tolerance to metals and drought
Eruca Sativa is a very versatile species grown worldwide despite drought
and disrupted soils, often grown along pathways and in dry fields
with limited water supply (Abassi et al., 2013). Furthermore it is a
species with capable tolerance and genetic ability to be tolerant of
44
Zn`s effect on health and growth of Eruca Sativa
metals as discussed above, however at very high contamination levels
it can succumb to toxicity.
Rapid growth
Eruca Sativa is an easy to grow and rapid species (Barlas et al., 2011).
These study further backs this up, this study was done through the
winter months so growth conditions were not perfect despite the
light and temperature being regulated in the greenhouse yet it took
only 49 days after being sown to reach its maximum size.
Resistant to disease.
Eruca Sativa suffered at the highest treatment level however this was
due to toxicity not because of disease. Eruca Sativa has been identified
as a species which has natural resistance to drought and pathogenic
invasion (Abassi et al., 2013)
Conclusion
Zn effect
45
Zn`s effect on health and growth of Eruca Sativa
It is possible that Zn concentration has a limited effect on
growth; however in high contamination levels Zn can have a
highly negative effect on growth, stunted growth and reduced
ability to bolt into flower are symptoms of Zn toxicity.
At the low treatment with 400 parts per million applied the
productivity and health of Eruca Sativa was at its greatest, this
suggest that Zn can have many positive effects and contributes
to the physiological mechanisms that can provide optimised
yield.
Treatments with higher contaminations caused reduced health and
productivity and signs of stress begin to show. The above 800
mg/kg contamination noticeable and significant symptoms of
toxicity began to kick in
Zn does not have an effect on the biomass production of Eruca
Sativa, but it does have an effect on the production of
chlorophyll, high chlorophyll content was measured at low level
treatments.
Eruca Sativa
Eruca sativa is not a hyper-accumulator of Zn, it does not
hyper accumulate Zn into harvestable biomass.
It is likely that it has the ability to tolerate Zn up to a
threshold in the same nature an HM excluder species would,
whereby below the threshold Zn uptake is low but once past the
46
Zn`s effect on health and growth of Eruca Sativa
threshold Zn begins to flood into the harvestable biomass and
severe toxicity begins.
Eruca Sativa is a very easy to grow, tolerant species that can
withstand a lot of stresses and still be healthy.
Critique
Not enough matter for separate compartment: one problem was that
separate Zn analysis for the leaf and shoot for the high treatment
was not possible so they needed to be mixed only allowing a total Zn
uptake.
Competition effected results: there were three individuals grown in
each pot and the seeds acquired were not homogenous therefore there
is the chance that competition has occurred throughout the growth
phase and altered the results.
The roots were not harvested: The roots were not harvested and
washed for Zn uptake analysis and for other observational purposes.
Had this been done translocation factor could have been considered
as well as previously reported symptoms of Zn deficiency and
toxicity on the root.
47
Zn`s effect on health and growth of Eruca Sativa
Weights were totals rather than leaves/shoots: a minor criticism of
the methodology separate shoot and leaf, fresh and dry weights were
not taken, the weights recorded were for the whole of the
harvestable portion.
Soil samples post-harvest: Another area in which this project could
have been improved with increased time frame for analysis would have
been post-harvest soil tests for Ph and other metal concentrations.
This was not done due to timing issues.
Flowering: Due to day length extending and the natural flowering
cycle of eruca sativa Meant many of the individuals flowered before
the harvesting stage; this is unlike practises for Eruca Sativa crop
growth and harvesting as it is commonly harvested prior to flowering
for use in saladsLack of physiological analysis: Due to lack of
resources and time many of the physiological effects of zinc could
not be analysed any more deeply than observations. Given an extended
time frame and greater resources many physiological factors such as
enzyme and hormone activity could have been analysed.
Sulphur effect on the growth: The compound used to create the
standard solution that the treatments were based on was zinc
sulphate. The fact that high levels of Zinc were required for
setting up treatments it Means that a high concentration of sulphate
will also be in the treatment applied., there was a plan to conduct
48
Zn`s effect on health and growth of Eruca Sativa
a parallel sulphur based experiment however it was not possible due
to lack of greenhouse and the time frame was too narrow to conduct
the studies one after the other. Furthermore the concentrations of
sulphur were not excessive; in fact it can be said that sulphur
toxicity is not likely as its presence reduces Ph which in itself
reduces the bioavailability of sulphur.
Further work
This study has provided additional framework to our knowledge of
heavy metal and species relationships. Expansion is greatly
necessary on what is already known of this relationship. There is a
huge field of work in this area and it is expanding at a relatively
rapid speed due to its importance scientifically and commercially.
Phyto exclusion and more generally Phytoremediation are incredible
exciting regions of biological science, and is one that is receiving
a lot of attention. The implications of further research down this
area could be huge and it is one that should be recommended greatly.
Pollution and soil contamination has become a real issue and a grand
talking point from many perspectives and dealing with the
repercussions of industrial boom and continued emissions of Heavy
metals is emerging as an area that is greatly in need of expansion.
Zinc is an essential micro nutrient and has received a lot of
attention from researchers. Zn is an essential component of health
49
Zn`s effect on health and growth of Eruca Sativa
and growth of plants knowledge on the topic can be further expanded.
Likewise there is also further work that can be done in the
physiological departments of heavy metals. Each Heavy metal is
essential to plants, each one having countless physiological roles.
Increasing the work in the field of Heavy metal and plant
interactions should be channelled through a more physiological
route, identifying Mechanism essential for crop survival or thriving
would be incredibly useful.
Eruca Sativa is clearly a species capable of tolerating harsh conditions
and has potentially got hyper-accumulative capabilities for other
heavy metals. This study suggests that its tolerance is due to its
genetic capabilities as a Heavy metal excluder.
Eruca Sativa is a commonly produced crop for consumption. Optimisation
of crop yield and increased growth speeds has many commercial
implications.
Hyper-accumulators are a relatively new research area and can always
be improved, their application can provide a coping device for
contaminated soil remediation an area that is clearly one that is
held to be of high importance.
As to the specifics of Zn and Eruca Sativa there are many expansions.
This study was a general over sight of the relationship with
inconclusive results in some areas. There are still a lot of
50
Zn`s effect on health and growth of Eruca Sativa
specifics to be looked into. This study was by no Means perfect and
the flaws identified in the critique can be ironed out, and the data
collected can be expanded in terms of numbers and specialised in
terms of the physiological effects.
Acknowledgments
I would like to thank Darrell Watts for all of his time and his
incredible support throughout the past 6 months.
Im also grateful for the help from Laura, James, jenny and Derek.
Their time and assistance was valued and from them I learnt a lot.
And lastly I’d like to Thank Nigel Chaffey for his guidance and
encouragement over the course of this process. His supervision was
much appreciated.
51
Zn`s effect on health and growth of Eruca Sativa
Thank you to my friends and family
References
Abbasi, B.H. Ali, J. Ali, M. Zia, M, Bokhari, S. Khan, M.A. (2013).
Free radical scavenging activity in in vitro-derived tissues of
52
Zn`s effect on health and growth of Eruca Sativa
Eruca sativa. Toxicology and Industrial Health, volume: unknown: 1-
8.[online]Available
from:http://www.researchgate.net/publication/256489379_Free_radical_
scavenging_activity_in_in_vitro-derived_tissues_of_Eruca_sativa/
file/9c9605238a83e777f4.pdf[accessed23/4/2014]
agricultural soils.
Alkorta, I. Hernanandez - Allica, J. Becerril, J.M. Amezaga, I.
Albizu, I, Garbisu, C. (2004). Recent findings on the
phytoremediation of soils contaminated with environmentally toxic
heavy metals and metalloids such as zinc, cadmium, lead, and
arsenic. Reviews in Environmental Science and Bio/Technology, Volume
3: 71-90.[online]Available
from:http://www.bioon.com/biology/UploadFiles/200412/200412291956158
44.pdf[accessed22/4/2014]
Allen, J.F. Forsberg, J. (2001) Molecular recognition in thylakoid
structure and function. TRENDS in Plant Science, Volume 6:317-325.
[online]Available
from:http://www.cell.com/trends/plant-science/abstract/S1360-
1385%2801%2902010-6?_returnURL=http%3A%2F%2Flinkinghub.elsevier.com
%2Fretrieve%2Fpii%2FS1360138501020106%3Fshowall%3Dtrue?
_returnURL=http%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii
%2FS1360138501020106%3Fshowall%3Dtrue[accessed8/4/2014]
53
Zn`s effect on health and growth of Eruca Sativa
Al-Qurainy, F. (2010) Application of inter simple sequence repeat
(ISSR marker) to detect genotoxic effect of heavy metals on Eruca
sativa (L.). African Journal of Biotechnology Volume 9(4): 467-474.
[online]Available from:http://www.google.co.uk/url?
sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0CDAQFjAA&url=http%3A%2F
%2Fwww.ajol.info%2Findex.php%2Fajb%2Farticle%2Fdownload
%2F77950%2F68346&ei=XfRsU4L1G9SZ0QWivoHgAQ&usg=AFQjCNGEY7jh3r_1bhw82
Y-
0mytgZDzZPA&sig2=pqdKJhAwcGMr6mfVnUWW2g&bvm=bv.66330100,d.d2k[access
ed//2014]
Ambrosone, C.B. Tang, L. (2009) Cruciferous Vegetable Intake and
Cancer Prevention: Role ofNutrigenetics. Cancer Prev Res, volume
2:298-300[online]Available
from:http://cancerprevention.aacrjournals.org/content/2/4/298.full[a
ccessed13/4/2014]
Ashraf, M. Harris, P.J.C. (2004) Potential biochemical indicators of
salinity tolerance in plants. Plant Science, volume 166: 3-16.
[online]Available
from:http://nhjy.hzau.edu.cn/kech/ssyy/qysd/njsl/21.pdf[accessed17/4
/2014]
Assuncao, A.G.L. Schat, H. Aaarts, M.G.M. (2010) Regulation of the
adaptation to zinc deficiency in plants. Plant Signaling &
54
Zn`s effect on health and growth of Eruca Sativa
Behavior , volume: 5(12): 1553-1555.[online]Available
from:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3115101/[accessed4/
5/2014]
Babula, P. Adam, V. Opatrilova, R. Zehnalek, J, Havel, L. Kizek, R.
(2008) Uncommon heavy metals, metalloids and their plant toxicity: a
review. Environ Chem Lett, volume 6:189-213[online]Available
from:http://pages.towson.edu/racasey/602docs/Sb_literature/Environ
%20Chem%20Lett%202008%20189-213.pdf[accessed6/5/2014]
Badr, N. Fawzy, M. Al- Qahtani, K.M. (2012) Phytoremediation: An
Ecological Solution to Heavy-Metal-Polluted Soil and Evaluation of
Plant Removal Ability. World Applied Sciences Journal, volume16 (9):
1292-1301.[online]Available
from:http://idosi.org/wasj/wasj16%289%2912/16.pdf[accessed12/4/2014]
Bakker, J.D. (2008) Increasing the utility of Indicator Species
Analysis. Journal of Applied Ecology, volume 45: 1829-1835.
[online]Available
from:http://onlinelibrary.wiley.com/doi/10.1111/j.1365-
2664.2008.01571.x/pdf[accessed13/4/2014]
Barlas, N.T. Irget, M.E. Tepecik, M. (2011) Mineral content of the
rocket plant (Eruca sativa). African Journal of Biotechnology Volume
10(64):14080-14082.[online]Available
55
Zn`s effect on health and growth of Eruca Sativa
from:http://www.ajol.info/index.php/ajb/article/view/96858[accessed/
/2014]
Beemster, G.T.S, Fiorani, F. Inze, D. (2003)Cell cycle: the key to
plant growth control?. tRENDS in Plant Science Volume.8(4): 154-
158[online]Available from:Beemster2003.pdf[accessed2/5/2014]
Bert, V. Macnair, M.R. De Laguerie, P. Saumitou - laprade, P. Petit,
D. (2000). Zinc tolerance and accumulation in metallicolous and
nonmetallicolous populations of Arabidopsis halleri (Brassicaceae).
New Phytol, volume 146: 225-233.[online]Available
from:http://gepv.univ-lille1.fr/downloads/downloads_labo/Bert%20et
%20al%202000%20%28New%20Phytol%29.pdf[accessed10/4/2014]
Broadley, M.R. White, P.J. Hammond, J.P. Zelko, I. Lux, A. (2007)
Zinc in plants. New Phytologist, volume: 173: 677-702.
[online]Available
from:http://onlinelibrary.wiley.com/doi/10.1111/j.1469-
8137.2007.01996.x/pdf[accessed1/5/2014]
Cag, S. Cevahir, G. Unal, M. Kaplan, E. Cingil, C. Kosesakal, T.
(2004) THE EFFECT OF Zn, Cu AND Mn ON SENESCENCE IN EXCISED
COTYLEDONS OF Eruca sativa L. Fresenius Environmental Bulletin,
Volume 13: 733-729.[online]Available
from:http://www.researchgate.net/publication/236893083_THE_EFFECT_OF
56
Zn`s effect on health and growth of Eruca Sativa
_Zn_Cu_AND_Mn_ON_SENESCENCE_IN_EXCISED_COTYLEDONS_OF_Eruca_sativa_L[
accessed17/4/2014]
Cakmak, I. (2000) Possible roles of zinc in protecting plant cells
from damage by reactive oxygen species. New Phytol, Volume 146: 185-
205.[online]Available
from:http://www.harvestzinc.org/pdf/ZincinProtectingPlantsfromFreeRa
dicals.pdf[accessed23/4/2014]
Cakmak. I. (2008) Enrichment of cereal grains with zinc: Agronomic
or genetic biofortification?. Plant Soil, Volume 302:1–
17[online]Available
from:http://zinc-crops.ionainteractive.com/publications/Enrichment.p
df[accessed17/4/2014]
Cobbet, C. Golsburgh, P. (2002) PHYTOCHELATINS ANDMETALLOTHIONEINS:
Roles in Heavy Metal Detoxification and Homeostasis. Annu. Rev.
Plant Biol, Volume 53:159–82[online]Available
from:http://www.plantstress.com/articles/toxicity_m/phytochelatins.p
df[accessed27/4/2014]
Cobbett, C. Goldsbrough, P. (2002) PHYTOCHELATINS
ANDMETALLOTHIONEINS: Roles in Heavy Metal Detoxification and
Homeostasis. Annu. Rev. Plant Biol, Volume 53:159–82.
[online]Available
from:http://www.annualreviews.org/doi/abs/10.1146/annurev.arplant.53
57
Zn`s effect on health and growth of Eruca Sativa
.100301.135154?url_ver=Z39.88-2003&rfr_dat=cr_pub
%3Dpubmed&rfr_id=ori%3Arid
%3Acrossref.org&journalCode=arplant[accessed17/4/2014]
Cobbett, C.S. (2000) Phytochelatins and Their Roles in Heavy Metal
Detoxification. Plant Physiology, Volume 123:825–
832[online]Available
from:http://www.plantstress.com/articles/toxicity_m/phytochelatins.p
df[accessed26/4/2014]
Dahmani-Muller, H. Van Oort, F. Gelie, B. Balabane, M. (2000)
Strategies of heavy metal uptake by three plant species growing near
a metal smelter. Environmental Pollution, volume 109: 231-
238[online]Available
from:http://www.researchgate.net/publication/8610545_Strategies_of_h
eavy_metal_uptake_by_three_plant_species_growing_near_a_metal_smelte
r/file/9c96051b8736551ef2.pdf[accessed23/4/2014]
Diancono, M. Montemurro, F. (2009) Long-term effects of organic
amendments on soil fertility. A review. Agron. Sustain. Dev. Volume
30: 401–422[online]Available
from:http://hal.archives-ouvertes.fr/docs/00/88/65/39/PDF/hal-
00886539.pdf[accessed23/4/2014]
Dolezalova, I. Duchoslav, M. Dusek, K. (2013) Biology and Yield of
Rocket (Eruca sativa Mill.) under Field Conditions of the Czech
58
Zn`s effect on health and growth of Eruca Sativa
Republic (Central Europe). Not Bot Horti Agrobo, Volume 41(2):530-
537[online]Available from:http://www.google.co.uk/url?
sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0CDEQFjAA&url=http%3A%2F
%2Fwww.notulaebotanicae.ro%2Findex.php%2Fnbha%2Farticle%2Fdownload
%2F9281%2F7665&ei=wfRsU_OSAo3Y0QXy74DYAw&usg=AFQjCNG8huFhTs8Pm0P2j6x
viwgm0pvh-g&sig2=cqaz-
XbMlC8doF83mSOqAA&bvm=bv.66330100,d.d2k[accessed//2014]
Duruibe, J.O, Ogwuegbu, M.O.C, Egwurugwu, J.N (2007)Heavy metal
pollution and human biotoxic effects. international Journal of
Physical Sciences, Volume 2 (5): 112-118, [online]Available
from:http://academicjournals.org/journal/IJPS/article-abstract/59CA3
5213127[accessed20/4/2014]
Eckert, D & sims, J.T. (2009) Recommended Soil pH and Lime
Requirement Tests. Recommended Soil Testing Procedures for the
North-eastern United States.
Ghosh, M. Singh, S.P. (2005) A Review on Phytoremediation of Heavy
Metals and Utilization of It’s by Products. As. J. Energy Env,
volume 6 (4): 214-231[online]Available
from:http://www.ecology.kee.hu/pdf/0301_001018.pdf[accessed10/4/2014
]
Grotz, N. Guerinot, M.L. (2006) Molecular aspects of Cu, Fe and Zn
homeostasis in plants. Biochimica et Biophysica Acta, volume
59
Zn`s effect on health and growth of Eruca Sativa
1763:595-608.[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0167488906001
157[accessed29/4/2014]
Hall, J.L. (2001) Cellular mechanisms for heavy metal detoxification
and tolerance. journal of experimental botany, Volume 53 (366): 1-
11.[online]Available
from:http://jxb.oxfordjournals.org/content/53/366/1.full.pdf+htmL[ac
cessed17/4/2014]
Hooda, V. (2005) Phytoremediation of toxic metals from soil and
waste water. Journal of Environmental Biology, Volume 28(2): 367-
376[online]Available
from:http://www.jeb.co.in/journal_issues/200704_apr07_supp/paper_04.
pdf[accessed20/4/2014]
Inskeep, W.P. Bloom, P.R. (1985) Extinction Coefficients of
Chlorophyll a and b in N,N-Dimethylformamide and 80% Acetone 1.
Plant Physiol. Volume: 77: 483-485.[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0167488906001
157[accessed3/5/2014]
Inskepp, W.P. Bloom, P.R. (1985) Extinction Coefficients of
Chlorophyll a and b in N,N-Dimethylformamide and 80% Acetone1.plant
Physiol, Volume 77: 483-485.[online]Available
from:http://www.plantphysiol.org/content/77/2/483[accessed25/4/2014]
60
Zn`s effect on health and growth of Eruca Sativa
Jarup, L. (2003) Hazards of heavy metal contamination. British
Medical Bulletin, volume68: 167-182[online]Available
from:http://bmb.oxfordjournals.org/content/68/1/167.full.pdf+htmL[ac
cessed11/4/2014]
Jaske, M. Hacin, J. Marsic, K. (2012) Production of rocket (Eruca
sativa Mill.) on plug trays and on a floating system in relation to
reduced nitrate content. Acta agriculturae Slovenica, volume 101:
59-68.[online]Available
from:http://aas.bf.uni-lj.si/marec2013/07Jakse
%20M..pdf[accessed9/4/2014]
Khan, A.G. Kuek, C. Chaundry, T.M. Khoo, C.S. Hayes, W.J. (2000)
Role of plants, mycorrhizae and phytochelators in heavy metal
contaminated land remediation. Chemosphere, Volume: 41: 197-207.
[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0045653599004
129[accessed13/4/2014]
Lasat, MM. (2002)Phytoextraction of Toxic Metals: A Review of
Biological Mechanisms. J. Environ. Qual, Volume (31): 109-
120[online]Available
from:http://www.epa.gov/tio/download/remed/lasat_article.pdf[accesse
d17/4/2014]
61
Zn`s effect on health and growth of Eruca Sativa
Lin, Y. Aarts, M.G.M. (2012) The molecular mechanism of zinc and
cadmium stress responsein plants. Cell. Mol. Life Sci, Volume
69(19): 3187-3206 [online]Available
from:http://www.academia.edu/2670488/The_molecular_mechanism_of_zinc
_and_cadmium_stress_response_in_plants[accessed12/4/2014]
Lone, M.I. He, Z. Yang, X. (2008)Phytoremediation of heavy metal
polluted soils and water: Progresses and perspectives. J Zhejiang Univ Sci
B. Mar, Volume: 9(3): 210-220. [online] available from:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2266886/#__ffn_sectitle
[acccessed: 15/4/2014].
Masestri, E, Marmiroli, M, Visioli, G, Marmiroli, N. (2010), Metal
tolerance and hyperaccumulation: Costs and trade-offs between traits
and environment. Environmental and Experimental Botany, volume (68):
1-13[online]Available
from:http://plantstress.com/Articles/up_toxicity_files/Metal
%20tolerance%20and%20hyperaccumulation%202010.pdf[accessed13/4/2014]
Matthiessen, M.K. Larney, F.J. Selinger, L.B. Olson, A.F. (2005)
Influence of Loss-on-Ignition Temperature and Heating Time on Ash
Content of Compost and Manure. Communications in Soil Science and
Plant Analysis, Volume: 36: 2561-2573.[online]Available
from:http://people.uleth.ca/~selibl/Brentspapers/Matthiessenetal2005
.pdf[accessed22/4/2014]
62
Zn`s effect on health and growth of Eruca Sativa
Mcgrath, S.P. Zhao, F. (2003) Phytoextraction of metals and
metalloids from contaminated soils. Current Opinion in
Biotechnology, volume 14: 277-282.[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0958166903000
600[accessed24/4/2014]
Mganga, N. Manoko, M.L.K. Rulangaranga, Z.K. (2011) CLASSIFICATION
OF PLANTS ACCORDING TO THEIR HEAVY METAL CONTENT AROUND NORTH MARA
GOLD MINE, TANZANIA:IMPLICATION FOR PHYTOREMEDIATION, tanz. J.Sci. Volume
37: 109-119. [online] available from:
http://www.ajol.info/index.php/tjs/article/viewFile/73619/63781
[acccessed: 15/4/2014]
Michael, H.N. Shafiki, R.E. Rasmy, G.E. (2011). Studies on the
chemical constituents of fresh leaf of Eruca sativa extract and its
biological activity as anticancer agent in vitro. Journal of
Medicinal Plants Research Volume: 5(7): 1184-1191.[online]Available
from:http://www.academicjournals.org/article/article1380533018_Micha
el%20et%20al.pdf[accessed21/4/2014]
Mousavi, S.R. (2011)Zinc in Crop Production and Interaction with
Phosphorus. Australian Journal of Basic and Applied Sciences, Volume
5(9): 1503-1509.[online]Available from:http://ijappjournal.com/wp-
content/uploads/2013/03/64-68.doc.pdf[accessed20/4/2014]
63
Zn`s effect on health and growth of Eruca Sativa
Mousavi, S.R. Galavi, M. Rezaei, M. (2013) Zinc (Zn) Importance for
Crop Production – A Review. International journal of Agronomy and
Plant Production. Volume 4(1):64-68.[online]Available
from:http://ijappjournal.com/wp-content/uploads/2013/03/64-
68.doc.pdf[accessed9/4/2014]
Myers, S.S. Zanobetti, A. Kloog, I. Huybers, P. Leakey, A.D.B.
Bloom, A.J et al. (2014). Increasing CO2 threatens human nutrition.
Nature, Volume: 1: 1-11.[online]Available
from:http://www.nature.com/nature/journal/vaop/ncurrent/full/nature1
3179.htmL[accessed21/4/2014]
Nagajyoti, P.C. Lee, K.D. Sreekanth, T.V.M (2010) Heavy metals,
occurrence and toxicity for plants: a review. Environ Chem Lett,
volume 8: 199-216.[online]Available
from:http://nhjy.hzau.edu.cn/kech/ssyy/pdf/hvmet/2.pdf[accessed3/5/2
014]
Neeto, A.T. Campostrini, E. Gonc, J. Oliviera, A.D. Bressan-Smith,
R.E. (2005) Photosynthetic pigments, nitrogen, chlorophyll a
fluorescence and SPAD-502 readings in coffee leaves. Scientia
Horticulturae, Volume 104: 199-209. [online] available
from:http://www.google.co.uk/url?
sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0CDAQFjAA&url=http%3A%2F
%2Fwww.researchgate.net%2Fpublication
64
Zn`s effect on health and growth of Eruca Sativa
%2F37621556_The_responses_of_plants_to_metal_toxicity._A_review_focu
sing_on_copper_manganese__zinc%2Ffile
%2F72e7e5167bc84e748e.pdf&ei=Tc9sU7-
YFsOAPafDgYgP&usg=AFQjCNHqq8i79j9iIKeqjAXuZDY5eViBHw&sig2=yh2ZrB5HNm
ARqwwo7kQjDw&bvm=bv.66330100,d.ZWU .[acccessed: 15/4/2014].
Oliviera, A. Pampulha, M.E. (2006) Effects of Long-Term Heavy Metal
Contamination on Soil Microbial Characteristics. JOURNAL OF
BIOSCIENCE AND BIOENGINEERING, Volume102(3): 157–
161[online]Available
from:https://www.jstage.jst.go.jp/article/jbb/102/3/102_3_157/_artic
le[accessed5/5/2014]
Ozdener, Y. Aydin, B.K. (2010) The effect of zinc on the growth and
physiological and biochemical parameters in seedlings of Eruca
sativa (L.) (Rocket). Acta Physiologiae Plantarum, Volume 32(3):469-476.
[online] available from:
http://link.springer.com/article/10.1007%2Fs11738-009-0423-z
[acccessed: 15/4/2014].
Padamavethiamma, P.K. Li, L.Y. (2007). Phytoremediation Technology:
Hyper-accumulation Metals in Plants. Water Air Soil Pollution,
volume 184:105-126[online]Available
from:http://www.researchgate.net/publication/225710092_Phytoremediat
65
Zn`s effect on health and growth of Eruca Sativa
ion_Technology_Hyper-accumulation_Metals_in_Plants/file/
3deec519b37aebd35b.pdf[accessed20/4/2014]
Pal, S. Patra, A.K, Reza, S.K, Wildi, W. Pote, J. (2010) Use of Bio-
Resources for Remediation of Soil Pollution. Natural Resources,
volume 1: 110-125[online]Available from: Use of Bio-Resources for
Remediation of Soil Pollution. [accessed//2014]
Palmer, C.M. Guerinot, M.L. (2009)Facing the challenges of Cu, Fe
and Zn homeostasis in plants. nature chemical biology, Volume 5
(5) :333-340[online]Available
from:http://www.nature.com/nchembio/journal/v5/n5/full/nchembio.166.
htmL[accessed20/4/2014]
Pilon-Smits, E. (2005) Phytoremediation. Annu. Rev. Plant Biol,
volume 56: 15-39[online]Available
from:http://www.annualreviews.org/doi/abs/10.1146/annurev.arplant.56
.032604.144214[accessed30/4/2014]
Porra, R.J. (2002) The chequered history of the development and use
of simultaneous equations for the accurate determination of
chlorophylls a and b. Photosynthesis Research, volume 73: 149–156.
[online]Available from:http://link.springer.com/article/10.1023%2FA
%3A1020470224740#page-1[accessed4/5/2014]
66
Zn`s effect on health and growth of Eruca Sativa
Prasad,M.N.V. Freitas, H.M.D.O. (2003) Metal hyperaccumulation in
plants - Biodiversity prospecting for phytoremediation technology.
Electron. J. Biotechnol. Volume 6(3): 285-321 [online] available from:
http://www.scielo.cl/scielo.php?pid=S0717-
34582003000300012&script=sci_arttext&tlng=e [acccessed: 15/4/2014].
Pulford, L.D. Watson, C. (2002) Phytoremediation of heavy metal-
contaminated land by trees—a review. Environment International,
Volume: 29: 529– 540.[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0160412002001
526[accessed//2014]
Rout, G.R. Das, P. (2003) Effect of metal toxicity on plant growth
and metabolism: I. Zinc. Agronomie, Volume 23: 3–11.
[online]Available
from:http://link.springer.com/chapter/10.1007%2F978-90-481-2666-
8_53#page-1[accessed26/4/2014]
RSC (undated) Atomic absorption spectrometry
Sagardoy, R. Morales, F. Lo` Pez-Millan, A.F. Abadia. A. Abadia. J.
(2009) Effects of zinc toxicity on sugar beet (Beta vulgaris L.)
plants grown in hydroponics. Plant Biology, Volume 11: 339-350.
[online]Available
from:http://onlinelibrary.wiley.com/doi/10.1111/j.1438-
8677.2008.00153.x/
67
Zn`s effect on health and growth of Eruca Sativa
abstract;jsessionid=93BFBF61C0DD4F3CB1740EC02FF730E3.f01t04[accessed
18/4/2014]
Saie, A. Karaipekli, A. (2008) Preparation, thermal properties and
thermal reliability of capric acid/expanded perlite composite for
thermal energy storage. Materials Chemistry and Physics, volume
109: 459 - 464.[online]Available from:http://www.google.co.uk/url?
sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0CDAQFjAA&url=http%3A%2F
%2Fwww.researchgate.net%2Fpublication
%2F232388629_Preparation_thermal_properties_and_thermal_reliability_
of_capric_acidexpanded_perlite_composite_for_thermal_energy_storage
%2Ffile
%2Fd912f50feee94d772e.pdf&ei=BPVsU8eUE4Sl0AXOyIH4AQ&usg=AFQjCNHucYo1
ZvAxXYq8Jk-M4CfmNvqjmg&sig2=sEnkxSpCTtDCNqrw48EbYA[accessed//2014]
Saleh, A.A.H. (2001) Effect of Cd and Pb on growht, certain
antioxidant enxymes activity, protien profile and accumulation of
Cd, Pb and Fe in raphanus sativas and eruca sativa saplings. Egyptian
journal of biology, Volume 3: 131-139. [online] available from:
http://ecology.nottingham.ac.uk/~plzfg/EBBSoc/ejb3/botany/Saleh_2001
.pdf [acccessed: 15/4/2014].
Sarma, H. (2011) Metal hyperaccumulation in plants: A review
focusing on phytoremediation technology. Journal of environmental
science and technology, Volume 4(2): 118-138.[online]Available
68
Zn`s effect on health and growth of Eruca Sativa
from:http://docsdrive.com/pdfs/ansinet/jest/2011/118-
138.pdf[accessed//2014]
Schmidt, U. (2003) Enhancing Phytoextraction: The Effect of Chemical
Soil Manipulation on Mobility, Plant Accumulation, and Leaching of
Heavy Metals. J. Environ. Qual, Volume: 32:1939–
1954[online]Available from:http://www.dzumenvis.nic.in/Microbes
%20and%20Soil%20Fertility/pdf/Enhancing
%20Phytoextraction.pdf[accessed7/5/2014]
Schmidt, U. (2003) Enhancing Phytoextraction: The Effect of Chemical
Soil Manipulation on Mobility, Plant Accumulation, and Leaching of
Heavy Metals. J. ENVIRON. QUAL, Volume 32: 1939-1954[online]
available from: http://www.dzumenvis.nic.in/Microbes%20and%20Soil
%20Fertility/pdf/Enhancing%20Phytoextraction.pdf [acccessed:
15/4/2014]
Schwart, C. Echevarria, G. Morel, J.L. (2003) Phytoextraction of
cadmium with Thlaspi caerulescens. Plant and Soil, Volume 249: 27–
35.[online]Available
from:http://link.springer.com/article/10.1023%2FA
%3A1022584220411#page-1[accessed19/4/2014]
Smical, A-I. Hotea, V. Oros, V. Juhasz, J. Pop, E. (2008) STUDIES ON
TRANSFER AND BIOACCUMULATION OF HEAVY METALS FROM SOIL INTO LETTUCE.
Environmental Engineering and Management Journal, Volume 7(5): 609-
69
Zn`s effect on health and growth of Eruca Sativa
615.[online]Available
from:http://omicron.ch.tuiasi.ro/EEMJ/pdfs/vol7/no5/20_Smical_A.pdf?
origin=publication_detail[accessed29/4/2014]
Stals, H. Inze, D. (2001) When plant cells decide to divide. TRENDS
in Plant Science, volume 6 (8): 333-387[online]Available
from:http://math.unife.it/lm.biomolecolare/insegnamenti/biologia-
molecolare-vegetale/materiale-didattico-aa-13-14/11-ciclo-
cellulare/cellcycle2.pdf[accessed2/5/2014]
Takahashi, S. Goldberg, H.A. Feeney, C.A. Karim, D.P. Farrell, M.
O`Leary, K. et al. (2006) Gas barrier properties of butyl
rubber/vermiculite nanocomposite coatings. Polymer , volume 47:
3083-3093.[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0032386106002
527[accessed26/4/2014]
Tandy, S. Bossart, K, Mueller, R. Ritschel, J. Hauser, L. Schulin, R
et al., (2004). Extraction of Heavy Metals from Soils Using
Biodegradable Chelatin Agents. Environ. Sci. Technol.Volume 38: 937-944.
[online] available from:
http://pubs.acs.org/doi/abs/10.1021/es0348750[acccessed: 15/4/2014].
Tangahu, B.V. Abdullah, S.R.S. Basri, H. Idris, M. Anuar, N.
Mukhlisin. (2011)A Review on HeavyMetals (As, Pb, and Hg) Uptake by
Plants through Phytoremediation. International Journal of Chemical
70
Zn`s effect on health and growth of Eruca Sativa
Engineering, volume 2011: 1-31.[online]Available
from:http://www.hindawi.com/journals/ijce/2011/939161/[accessed15/4/
2014]
Tian, S. Lu, L. Yang, X. Labavitch, J.M. Huang, Y. Brown, P. (2009)
Stem and leaf sequestration of zinc at the cellular level in the
hyperaccumulator Sedum alfredii. New Phytologist, volume 182: 116–
126.[online]Available
from:http://onlinelibrary.wiley.com/doi/10.1111/j.1469-
8137.2008.02740.x/abstract[accessed14/4/2014]
Vamerali, T. Bandiera, M. Mosca, G. Field crops for phytoremediation
of metal-contaminated land. A review. Environ Chem Lett, volume 8:1–
17.[online]Available
from:http://www.leam.illinois.edu/sustrantoul/data/Vamerali%20et
%20al%202010%20-%20Field%20crops%20for%20phytoremediation%20of
%20metal-contaminated%20land%20-%20a%20review.pdf[accessed24/4/2014]
Vangronsveld, J. Herzig, R. Weyens, N. Boulet, J, Adrianaensen, K.
Ruttens, A et al. (2009) Phytoremediation of contaminated soils and
groundwater: lessons from the field. Environ Sci Pollut Res, volume
16: 765 - 794.[online]Available
from:http://www.ask-force.org/web/Environment/Vangronsveld-
Phytoremediation-Contaminated-Soils-2001.pdf[accessed13/4/2014]
71
Zn`s effect on health and growth of Eruca Sativa
Williams, L.E. Pittman, J.K. Hall, J.L. (2000) Emerging mechanisms
for heavy metal transport in plants. Biochimica et Biophysica Acta,
Volume 1465:104-126[online]Available
from:http://www.sciencedirect.com/science/article/pii/S0005273600001
334[accessed27/4/2014]
Wuana, R.A. Okiemen, F.E. (2011) HeavyMetals in Contaminated Soils:
A Review of Sources, Chemistry, Risks and Best Available Strategies
for Remediation. International Scholarly Research Network, volume
2011: 1-20.[online]Available
from:http://www.hindawi.com/journals/isrn/2011/402647/[accessed3/5/2
014]
Zazimalova, E. Napier, R.M. (2003) Points of regulation for auxin
action. Plant Cell Rep, Volume 21: 625-634.[online]Available
from:http://link.springer.com/article/10.1007%2Fs00299-002-0562-
9#page-1[accessed18/4/2014]
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Appendix
1. Randomizing table
2. Soil and water tests resulta
3. Metal calculations.
4. Tables of results
5. Anova and tukeys HSD readings
6. Kruskal wallis readings
7. T test readings
8. Boxplots and individuals plots
9. Risk assessment
73
Zn`s effect on health and growth of Eruca Sativa
Randomizing table
400b10 800b3 1200b8 400a4 400a8
1200a7 400a9 Ta6 400b1 400b2
400b6 800a5 Tb6 1200b4 800a2
1200a1 800a1 1200a6 1200a2 1200b6
Ta7 Ta5 1200b2 400b8 1200b5
1200a8 800b10 Ta2 Tb10 400b9
Ta4 Ta8 1200b7 400a6 400a2
800b2 800a4 800b9 Ta1 400b7
400b4 800b1 1200a9 Tb5 Ta10
400b3 1200a4 800b5 800a3 Ta3
400a5 Tb7 800b7 800a8 1200b1
400a1 1200a5 Tb2 400a7 800a7
1200a10 400a3 400a10 800a9 Tb4
Tb8 1200b9 800b8 Tb1 1200b3
Tb3 800a10 800b4 400b5 1200b10
800a6 Ta9 800b6 Tb9 1200a3
Table 4: Positioning of samples on the greenhouse bench. The rocket
was placed on a bench as close to the growing lights. Labels =
Treatment (t, 400, 800, 1200) – analysis group (A or B) - sample
number (1-10)
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Zn`s effect on health and growth of Eruca Sativa
Soil samples
Ph – 5.5
Organic matter – 45%
Water samples
Samples of The water used for watering the plants throughout the
growth phase was taken, each of the 10 samples came out with 0mg/g
of zinc content.
Metal calculations for treatments
ZnSO4. 7H2O
RMM 287.56
RAM Zn 65.4
% ZN 22.75
Weight of compound required for
100mg/kg Zn (1l dilution)
439.5g
Weight of compound required for
100mg/kg Zn (500mL dilution)
219.8g
75
Zn`s effect on health and growth of Eruca Sativa
RAM S 32.1
% S 11%
mg/kg S per 1000 Zn 111.5
mg/kg S per 1 Zn 0.11
Table 5: Zn sulphate treatment calculations
Treatment (mg/kg) ML`s of solution applied for
treatment
control 0
400 (low) 4
800 (medium) 8
1200 (high) 12
Table 6: Treatments calculations
Table 7: individual dry weights
Dry weight (g)
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Zn`s effect on health and growth of Eruca Sativa
Sample control 400 800 1200
1 1.59 1.99 No data No data
2 2.27 1.75 3.83 1.17
3 1.29 2.99 1.44 1.35
4 1.93 1.77 2.46 No data
5 2.57 2.16 1.52 1.17
6 2.32 1.76 3.45 1.20
7 1.45 2.17 3.30 0.73
8 1.07 3.48 1.49 0.84
9 1.41 2.57 1.54 No data
10 1.31 1.96 2.29 No data
Number 10.00 10.00 9.00 6.00
Mean 1.72 2.26 2.37 1.08
Median 1.52 2.08 2.29 1.17
ST dev.0.4884070.553151 0.896394 0.217565
95% CIUpper 2.07 2.66 3.06 1.30
Lower 1.37 1.86 1.68 0.85
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Zn`s effect on health and growth of Eruca Sativa
Table 8: individual fresh weights
Fresh weight (g)
Sample control 400 800 1200
1 9.58 9.38 No data No data
2 11.72 8.14 17.12 3.48
3 7.85 15.25 9.07 5.24
4 10.77 7.3 11.89 No data
5 13.65 4.32 6 7.62
6 12.06 7.69 15.67 7.71
7 8.38 10.83 19.47 6.54
8 6.66 16.8 7.7 4.22
9 7.83 11.94 12.83 No data
10 6.62 11.36 13.31 No data
Number 10.00 10.00 9.00 6.00
Mean 9.51 10.30 12.56 5.80
Median 8.98 10.11 12.83 5.89
ST dev.2.3118863.576746 4.191676 1.617636
95% CIUpper 11.17 12.86 15.78 7.50
Lower 7.86 7.74 9.34 4.10
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Zn`s effect on health and growth of Eruca Sativa
Table 9: individual leaf Zn content
Leaf Zn concentration (mg/kg)
Samplecontrol 400.00800.00 1200.00
1 41.67 464.65 1041.67 No data
2 68.97 705.88 2172.04 No data
3 20.20 644.44 914.89No data
4 20.41 765.96 469.39No data
5 21.74 357.89 500.00No data
6 31.25 350.52 659.79No data
7 22.99 333.33 577.78No data
8 26.67 721.65 1409.09 No data
9 440.00 758.62No data
10 No data
N 8.00 9.00 10.00 0.00
Mean 31.74 531.59 944.81No data
Median24.83 464.65 758.62No data
79
Zn`s effect on health and growth of Eruca Sativa
SDEV 15.61 166.43 517.09No data
95% CIlower 18.69 403.66 574.91No data
Upper 44.78 659.52 1314.71 No data
Table 10: individual shoot Zn content
Shoot Zn concentration (mg/kg)
Samplecontrol 400.00800.00 1200.00
1 20.62 324.32 363.64No data
2 26.32 142.86 1025.00 No data
3 25.00 116.28 1747.13 No data
4 40.82 160.92 1454.55 No data
5 23.53 363.64 512.82No data
6 29.41 148.94 215.05No data
7 50.63 166.67 480.00No data
8 75.00 231.88 623.38No data
9 333.33 342.86No data
10 No data
80
Zn`s effect on health and growth of Eruca Sativa
N 8.00 9.00 9.00 0.00
Mean 36.42 220.98 751.60No data
Median27.86 166.67 512.82No data
SDEV 17.35 89.81 506.91No data
95% CIlower 21.91 151.95 361.95No data
Upper 50.92 290.01 1141.25 No data
Table: 11 individual total Zn uptakes
total Zn concentration (mg/kg)
samplecontrol 400.00800.00 1200.00
1 15.57 197.24 351.332946.24
2 23.82 212.18 799.264750.00
3 11.30 190.18 665.514901.10
4 15.31 231.72 480.985402.30
5 11.32 180.38 253.214204.08
6 15.17 124.86 218.714857.14
7 18.41 125.00 264.445043.48
8 25.42 238.38 508.124978.72
81
Zn`s effect on health and growth of Eruca Sativa
9 193.33 275.374240.00
10
N 8.00 9.00 9.00 9.00
Mean 17.04 188.14 424.104591.45
Median15.44 193.33 351.334857.14
SDEV 4.90 38.25 192.53681.80
95% CIlower 12.94 158.74 276.114067.37
upper 21.14 217.54 572.095115.53
Table 12: individual chlorophyll A content
Chlorophyll A
sample trace 400 800 1200
1 1.510453.26285 2.4558 1.648252 1.1756 2.4895 1.415 1.53753 2.736051.8897 1.8744 3.10614 0.759051.5176 1.32005 1.49315 1.1317 3.4573 1.3476 0.70955
6 2.2695 3.94275 0.9806 1.418557 2.0847 4.8141 2.06438 1.3471 3.05405 2.03019 1.627852.8718 1.590610 1.8259
N 9 10 9 6mean 1.626889 2.912555 1.675383 1.652175median 1.510452.962925 1.5906 1.5153SDEV 0.591292 0.973151 0.436801 0.71827295% CIlower 2.081396 3.608706 2.011138 2.405956
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Zn`s effect on health and growth of Eruca Sativa
upper 1.172382 2.216404 1.339629 0.898394
Table13: individual chlorophyll B content
Chlorophyll B
sample trace 400 800 1200
1 0.571151.16105 0.7323 0.400192 0.551060.83452 0.59012 0.541023 0.909290.51558 0.63744 0.741684 0.255110.60686 0.51245 0.616685 0.530080.90952 0.38568 0.371396 0.826261.11999 0.43634 0.376537 0.6303 1.40724 0.792788 0.482320.87827 0.78729 0.562670.662 0.6740410 0.56044
N 9 10 9 6mean 0.590916 0.865547 0.616483 0.507915median 0.562670.856395 0.63744 0.470605SDEV 0.1793 0.277933 0.139296 0.1384995% CIlower 0.728738 1.064369 0.723556 0.653252upper 0.453093 0.666725 0.509411 0.362578
Table 14: individual total chlorophyll (A+B) contentChlorophyll A+B total
Sample trace 400 800 1200
1 2.085684.43269 3.19467 2.052822 1.729873.33071 2.00896 2.082663 3.652692.41032 2.51688 3.856034 1.0162 2.12857 1.83607 2.113835 1.664874.37603 1.73688 1.082896 3.101885.07327 1.41961 1.798867 2.720586.23421 2.862668 1.833053.94048 2.822799 2.1949 3.54142 2.2689610 2.39123
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Zn`s effect on health and growth of Eruca Sativa
N 9 10 9 6Mean 2.222191 3.785893 2.296387 2.164515Median 2.085683.74095 2.26896 2.06774SDEV 0.763529 1.236651 0.560787 0.83532395% CI Lower 2.809091 4.67054 2.727446 3.041132Upper 1.635291 2.901246 1.865328 1.287898
Chlorophyll A/B ratio
Sample trace 400 800 1200
1 2.644577 2.810258 3.353544 4.1186692 2.133343 2.983152 2.397817 2.8418543 3.008996 3.665193 2.940512 4.1879254 2.975383 2.500742 2.575959 2.4211915 2.134961 3.801236 3.494088 1.9105256 2.746714 3.520344 2.24733 3.7674297 3.307473 3.420952 2.6038758 2.792959 3.477348 2.5788879 2.893081 4.338066 2.35980110 3.257976
N 9 10 9 6Mean 2.737499 3.377527 2.727979 3.207932Median 2.792959 3.44915 2.578887 3.304642SDEV 0.368062 0.497748 0.4163 0.8697895% CI lower 3.020416 3.733594 3.047976 4.12071Upper 2.454581 3.021459 2.407983 2.295154
ANOVA and tukeys HSD readings
One-way ANOVA: dry weight versus treatment
Source DF SS MS F PTreatment 3 7.651 2.550 6.10 0.002Error 31 12.961 0.418Total 34 20.612S = 0.6466 R-Sq. = 37.12% R-Sq. (ad) = 31.04%
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Zn`s effect on health and growth of Eruca Sativa
Individual 95% CIs for Mean Based onPooled StepLevel N Mean Step -+---------+---------+---------+--------0 10 1.7190 0.5148 (------*------)400 10 2.2599 0.5831 (------*------)800 9 2.3688 0.9508 (------*-------)1200 6 1.0752 0.2383 (--------*--------)-+---------+---------+---------+--------0.60 1.20 1.80 2.40
Pooled Step = 0.6466Grouping Information Using Tukey Methodtreatment N Mean Grouping800 9 2.3688 A400 10 2.2599 A0 10 1.7190 A B1200 6 1.0752 B
Means that do not share a letter are significantly different.
Tukey 95% Simultaneous Confidence IntervalsAll Pairwise Comparisons among Levels of treatment
Individual confidence level = 98.93%
treatment = 0 subtracted from:
treatment Lower Center Upper --------+---------+---------+---------+-400 -0.2443 0.5409 1.3261 (------*-----)800 -0.1569 0.6498 1.4565 (-----*------)1200 -1.5505 -0.6439 0.2628 (-------*------)--------+---------+---------+---------+--1.2 0.0 1.2 2.4
treatment = 400 subtracted from:
treatment Lower Center Upper --------+---------+---------+---------+-800 -0.6978 0.1089 0.9156 (------*------)1200 -2.0914 -1.1848 -0.2781 (------*-------)--------+---------+---------+---------+--1.2 0.0 1.2 2.4
treatment = 800 subtracted from:
treatment Lower Center Upper --------+---------+---------+---------+-1200 -2.2190 -1.2937 -0.3683 (------*-------)--------+---------+---------+---------+--1.2 0.0 1.2 2.4
One-way ANOVA: fresh weight versus treatment
Source DF SS MS F Ptreatment 3 167.7 55.9 4.88 0.007Error 31 355.2 11.5
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Zn`s effect on health and growth of Eruca Sativa
Total 34 522.9
S = 3.385 R-Sq = 32.07% R-Sq(adj) = 25.49%
Individual 95% CIs For Mean Based on Pooled StDevLevel N Mean StDev +---------+---------+---------+---------0 10 9.512 2.437 (-------*------)400 10 10.301 3.770 (------*-------)800 9 12.562 4.446 (-------*-------)1200 6 5.802 1.772 (--------*---------)+---------+---------+---------+---------3.0 6.0 9.0 12.0
Pooled StDev = 3.385
Grouping Information Using Tukey Method
treatment N Mean Grouping800 9 12.562 A400 10 10.301 A B0 10 9.512 A B1200 6 5.802 B
Means that do not share a letter are significantly different.
Tukey 95% Simultaneous Confidence IntervalsAll Pairwise Comparisons among Levels of treatment
Individual confidence level = 98.93%
treatment = 0 subtracted from:
treatment Lower Centre Upper ---------+---------+---------+---------+400 -3.321 0.789 4.899 (------*------)800 -1.173 3.050 7.273 (------*------)1200 -8.457 -3.710 1.036 (-------*-------)---------+---------+---------+---------+-6.0 0.0 6.0 12.0
treatment = 400 subtracted from:
treatment Lower Centre Upper ---------+---------+---------+---------+800 -1.962 2.261 6.484 (------*------)1200 -9.246 -4.499 0.247 (-------*------)---------+---------+---------+---------+-6.0 0.0 6.0 12.0
treatment = 800 subtracted from:
treatment Lower Centre Upper ---------+---------+---------+---------+1200 -11.605 -6.761 -1.916 (-------*-------)---------+---------+---------+---------+-6.0 0.0 6.0 12.0
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Zn`s effect on health and growth of Eruca Sativa
One-way ANOVA: Porro A versus pot
Source DF SS MS F Ppot 3 11.238 3.746 6.45 0.002Error 30 17.430 0.581Total 33 28.667
S = 0.7622 R-Sq = 39.20% R-Sq(adj) = 33.12%
Individual 95% CIs For Mean Based onPooled StDevLevel N Mean StDev ---+---------+---------+---------+------0 9 1.6269 0.6272 (--------*--------)400 10 2.9126 1.0258 (--------*-------)800 9 1.6754 0.4633 (--------*--------)1200 6 1.6522 0.7868 (----------*---------)---+---------+---------+---------+------1.20 1.80 2.40 3.00
Pooled StDev = 0.7622
Grouping Information Using Tukey Method
pot N Mean Grouping400 10 2.9126 A800 9 1.6754 B1200 6 1.6522 B0 9 1.6269 B
Means that do not share a letter are significantly different.
Tukey 95% Simultaneous Confidence IntervalsAll Pairwise Comparisons among Levels of pot
Individual confidence level = 98.93%
pot = 0 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+400 0.3322 1.2857 2.2391 (-------*-------)800 -0.9297 0.0485 1.0267 (-------*--------)1200 -1.0684 0.0253 1.1189 (--------*--------)---------+---------+---------+---------+-1.2 0.0 1.2 2.4
pot = 400 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+800 -2.1906 -1.2372 -0.2838 (-------*-------)1200 -2.3319 -1.2604 -0.1888 (-------*--------)---------+---------+---------+---------+-1.2 0.0 1.2 2.4
pot = 800 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+
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Zn`s effect on health and growth of Eruca Sativa
1200 -1.1169 -0.0232 1.0704 (--------*--------)---------+---------+---------+---------+-1.2 0.0 1.2 2.433 1.9723
S = 0.2123 R-Sq = 31.47% R-Sq(adj) = 24.62%One-way ANOVA: Porro B versus pot
Individual 95% CIs For Mean Based onPooled StDevLevel N Mean StDev ---+---------+---------+---------+------0 9 0.5909 0.1902 (-------*------)400 10 0.8655 0.2930 (------*------)800 9 0.6165 0.1477 (------*------)1200 6 0.5079 0.1517 (-------*--------)---+---------+---------+---------+------0.40 0.60 0.80 1.00
Pooled StDev = 0.2123
Grouping Information Using Tukey Method
pot N Mean Grouping400 10 0.8655 A800 9 0.6165 A B0 9 0.5909 B1200 6 0.5079 B
Means that do not share a letter are significantly different.
Tukey 95% Simultaneous Confidence IntervalsAll Pairwise Comparisons among Levels of pot
Individual confidence level = 98.93%
pot = 0 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+400 0.0091 0.2746 0.5401 (-------*------)800 -0.2468 0.0256 0.2980 (-------*-------)1200 -0.3875 -0.0830 0.2215 (--------*-------)---------+---------+---------+---------+-0.35 0.00 0.35 0.70
pot = 400 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+800 -0.5146 -0.2491 0.0164 (-------*------)1200 -0.6560 -0.3576 -0.0592 (--------*-------)---------+---------+---------+---------+-0.35 0.00 0.35 0.70
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Zn`s effect on health and growth of Eruca Sativa
pot = 800 subtracted from:
pot Lower Centre Upper ---------+---------+---------+---------+1200 -0.4131 -0.1086 0.1960 (--------*--------)---------+---------+---------+---------+-0.35 0.00 0.35 0.70Error 30 27.557 0.919Total 33 44.587
S = 0.9584 R-Sq = 38.20% R-Sq(adj) = 32.02%
One-way ANOVA: Porro total versus pot
Individual 95% CIs For Mean Based onPooled StDevLevel N Mean StDev ---+---------+---------+---------+------0 9 2.2222 0.8098 (-------*-------)400 10 3.7859 1.3035 (------*-------)800 9 2.2964 0.5948 (-------*-------)1200 6 2.1645 0.9151 (---------*---------)---+---------+---------+---------+------1.60 2.40 3.20 4.00
Pooled StDev = 0.9584
Grouping Information Using Tukey Method
pot N Mean Grouping400 10 3.7859 A800 9 2.2964 B0 9 2.2222 B1200 6 2.1645 B
Means that do not share a letter are significantly different.
Tukey 95% Simultaneous Confidence IntervalsAll Pairwise Comparisons among Levels of pot
Individual confidence level = 98.93%
pot = 0 subtracted from:
pot Lower Centre Upper +---------+---------+---------+---------400 0.3649 1.5637 2.7625 (-------*-------)800 -1.1558 0.0742 1.3042 (-------*--------)1200 -1.4328 -0.0577 1.3175 (---------*--------)+---------+---------+---------+----------3.0 -1.5 0.0 1.5
pot = 400 subtracted from:
pot Lower Centre Upper +---------+---------+---------+---------
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Zn`s effect on health and growth of Eruca Sativa
800 -2.6883 -1.4895 -0.2907 (-------*-------)1200 -2.9687 -1.6214 -0.2740 (--------*--------)+---------+---------+---------+----------3.0 -1.5 0.0 1.5
pot = 800 subtracted from:
pot Lower Centre Upper +---------+---------+---------+---------1200 -1.5070 -0.1319 1.2433 (--------*--------)+---------+---------+---------+----------3.0 -1.5 0.0 1.5
Kruskal-wallis readings
Kruskal-Wallis Test: Leaf Zn uptake versus treatment
Kruskal-Wallis Test on Leaf Zn uptake
treatment N Median Ave Rank Z
0 8 24.83 4.5 -4.00
400 9 464.65 14.8 0.62
800 9 758.62 20.2 3.26
Overall 26 13.5
H = 18.28 DF = 2 P = 0.000
Kruskal-Wallis Test: Shoot Zn Uptake versus treatment
Kruskal-Wallis Test on Shoot Zn Uptake
treatment N Median Ave Rank Z
0 8 27.86 4.5 -4.00
400 9 166.67 13.6 0.05
800 9 512.82 21.4 3.83
Overall 26 13.5
H = 20.65 DF = 2 P = 0.000
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Zn`s effect on health and growth of Eruca Sativa
H = 20.66 DF = 2 P = 0.000 (adjusted for ties)
Kruskal-Wallis Test: Zn uptake versus treatment
Kruskal-Wallis Test on Zn uptake
treatment N Median Ave Rank Z
0 8 15.44 4.5 -4.24
400 9 193.33 13.2 -1.62
800 9 351.33 21.8 1.28
1200 9 4857.14 31.0 4.42
Overall 35 18.0
H = 31.55 DF = 3 P = 0.000
T-test readings
Two-Sample T-Test and CI: 400 shoot, 400 leaf
Two-sample T for 400 shoot vs. 400 leaf
SE
N Mean StDev Mean
400 shoot 9 221.0 95.3 32
400 leaf 9 532 177 59
Difference = mu (400 shoot) - mu (400 leaf)
Estimate for difference: -310.6
95% CI for difference: (-456.3, -164.9)
T-Test of difference = 0 (vs. not =): T-Value = -4.65 P-Value = 0.001 DF = 12
Two-Sample T-Test and CI: control shoot, control leaf
Two-sample T for control shoot vs. control leaf
N Mean StDev SE Mean
control shoot 8 36.4 18.5 6.6
control leaf 8 31.7 16.7 5.9
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Zn`s effect on health and growth of Eruca Sativa
Difference = mu (control shoot) - mu (control leaf)
Estimate for difference: 4.68
95% CI for difference: (-14.37, 23.73)
T-Test of difference = 0 (vs. not =): T-Value = 0.53 P-Value = 0.605 DF = 13
Two-Sample T-Test and CI: 800 shoot, 800 leaf
Two-sample T for 800 shoot vs. 800 leaf
N Mean StDev SE Mean
800 shoot 9 752 538 179
800 leaf 9 945 548 183
Difference = mu (800 shoot) - mu (800 leaf)
Estimate for difference: -193
95% CI for difference: (-739, 352)
T-Test of difference = 0 (vs. not =): T-Value = -0.75 P-Value = 0.462 DF = 15
Boxplots and individual plots
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Zn`s effect on health and growth of Eruca Sativa
Figure 16: boxplot of chlorophyll A content between treatments
Figure 17: individual plot of chlorophyll A content between
treatments
Figure 16: boxplot of chlorophyll A content between treatments
93
Zn`s effect on health and growth of Eruca Sativa
Figure 18: boxplot of chlorophyll B content between treatments
Figure 19: Individual plot of chlorophyll B content between
treatments
94
Zn`s effect on health and growth of Eruca Sativa
Figure 20: boxplot of chlorophyll total content between treatments
Figure 21: individual plot of chlorophyll total content between
treatments
95
Zn`s effect on health and growth of Eruca Sativa
Figure 22: boxplot of fresh weights between treatments
Figure 23: individual plot of fresh weights between treatments
96
Zn`s effect on health and growth of Eruca Sativa
Figure 24: boxplot of dry weights between treatments
Figure 25: individual plot of dry weights between treatments
97
Zn`s effect on health and growth of Eruca Sativa
Figure 26: Boxplot of chlorophyll ratios
Figure 26: individual plots of chlorophyll ratios
98
Zn`s effect on health and growth of Eruca Sativa
SCHOOL OF SOCIETY, ENTERPRISE & ENVIRONMENT
RISK ASSESSMENT
STUDENT LABORATORY WORK
The purpose of this risk assessment is to identify potential hazards
associated with the proposed laboratory work, and to identify control
measures that eliminate or minimise risk to an acceptable level.
99
Zn`s effect on health and growth of Eruca Sativa
Notes for completion of Summary:
Students should complete this Summary page after identifying hazards and
appropriate controls in Table A and rating the level of risk using
Table B. If you are working alone you must also complete Table C.
SUMMARY OF RISK ASSESSMENT
Student name: Ryan Pullen
Student number: 228329
Module Code: BY6001
Date(s) of laboratory work:
from 4th Nov., 2013, in TE
and WE labs and N Park
greenhouse
Risk
Assessment ID number:
(office use)
Proposed laboratory work:
soil preparation, heavy metal uptake
measurements, fresh weigh, dry weight
measurements, chlorophyll extraction
100
Zn`s effect on health and growth of Eruca Sativa
Level of Risk
Low
(delete as applicable after completing
table A and consulting table B below)
Student signature
and date Ryan, Oct
2013
Supervising tutor signature
and date
Area Safety Manager
signature and date
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Zn`s effect on health and growth of Eruca Sativa
Table A Details of Risk Assessment
Notes for completion of Table A:
NB Separate risk assessments must be completed for each type of
activity planned.
Hazards: list significant hazards, for example: chemicals and
chemical reactions, fire or explosions, biohazards, radiation,
noise, high/low temperatures, electricity, radiation; or hazards
arising from the use of equipment, machinery and glassware.
Who might be harmed? Identify groups of people at risk from the
identified hazards, such as staff, students, general public, other?
Column 3 Controls: list precautions that are necessary to control
hazards, for example: proper labelling, storage and containment of
chemicals, provision and use of personal protective equipment, clear
instructions and training for handing chemicals and using equipment,
etc. Is First Aid provision adequate? Are particular measures
necessary to protect disabled or vulnerable colleagues?
Comments/Further action: are there other issues or concerns, which
require further action to reduce risk to an acceptable level?
Lone working should be avoided wherever possible. Will you be
working alone? Yes – in greenhouse (delete as appropriate). If yes
state under Comments/further action how you will be able to summon
help if an accident or emergency arises and complete Table C.
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Zn`s effect on health and growth of Eruca Sativa
Hazards Who might
be harmed?
Controls Comments/
further action
Zinc sulphates
or sulphur
compounds used
to level sulphur
mg/kg
Me, anyone
else
working
nearby.
Lab coat, goggles,
gloves, face mask
Prepare with care.
Observe usual
laboratory best
practice, e.g. no
hand-to-mouth
operations, wash
hands before
eating…
Acetone for
chlorophyll
extraction
Me, anyone
else
working
nearby
Lab coat, goggles,
gloves, face mask.
Fume cupboard.
Take care whilst
using the solvent.
Ensure room used
is well-
ventilated.
Zinc
contaminated
plant material
Me, anyone
else who
uses or
enters the
greenhouse/
anyone in
contact
Lab coats, gloves.
Face mask and
goggles when
grinding dried plant
material
Make sure anyone
entering the
greenhouse is
aware of the risks
103
Zn`s effect on health and growth of Eruca Sativa
with the
soil or
plant
Heating block
for zinc uptake
digestion stage
Me , anyone
in the room
whilst it
is on
Make sure to turn
it off when it
isn’t in use
Nitric acid Me, anyone
else
working
nearby
Lab coat, goggles,
gloves, face mask.
Fume cupboard to
contain fumes.
Use with great
care
Zinc
contaminated
soil, dried
plant material,
and solutions
Me, anyone
else
working
nearby
Ensure al is
appropriately
disposed of at end
of project.
Greenhouse work,
slips, trips
etc.
me Be careful and tidy Have phone on me
at all times so I
can contact anyone
if anything
happens
Table B Level of Risk
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Zn`s effect on health and growth of Eruca Sativa
Notes for completion of Table B:
Use table B to assess the level of risk (high, medium, low) by
estimating the likelihood of the identified hazards occurring, then
the potential severity of the consequences. If the activity is
assessed to be high risk action must be taken to reduce it to an
acceptable level (medium or low) – this should be discussed with the
Area Safety Manager if necessary.
Severity of
consequences
Likelihood
of hazard
Minor
injury/
modest
damage
Significant
accident/signif
icant damage
Major injury or
death/
major destructive
damage
Probably will occur Medium
Risk
High Risk High Risk
Possibly will occur Low Risk Medium Risk High Risk
Unlikely to occur Low Risk Low Risk Medium Risk
Table C: Working alone - only do so if it unavoidable.
In the event of an accident or emergency you must be contactable and
be able to contact others from where you will be working. You must
provide this information on each occasion that you work alone. It is
105
Zn`s effect on health and growth of Eruca Sativa
particularly important that a responsible adult is willing to act on
your behalf in an emergency or if you do not log off – provide
details below:
I agree to notify my contact with a log off time on each occasion I
work alone
Student signature: Ryan Pullen
2. Details of responsible adult (e.g. parent /partner/ friend)
Notes: Thank you for agreeing to act as a point of contact. By
doing so you are indicating your willingness to take appropriate
action on behalf of the student in the event of an emergency, or if
s/he does not log off. In practice this Means: if s/he has not
logged off 15 minutes after the scheduled time, you must try to
contact her/him at 10 minute intervals; if, after one hour, there is
no response then you must begin a search – this might include
contacting the emergency services.
Name of contact: Steven Pullen (Father)
Phone number or other Means of contact: home phone:
106
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