GOVERNMENT COLLEGE UNIVERSITY FAISALABAD

Department of Botany
Synopsis for MS
Title: Zinc-lysine mediated alterations in morpho-physiological and
ionomics of sunflower (Helianthus annuus L.)under cadmium stress
Name of Student: Farwa Jabeen

Registration No: GCUF-000747

ABSTRACT

A pot experiment will be conducted in the research area Government College University,
Faisalabad in order to determine the interactive effects of different levels of cadmium and zinc-
lysine. Two selected sunflower cultivars NK-278 and 5551 will be grown in soil and after
germination plants will be supplemented with different concentrations of CdCl2(0,20,40,60
CdCl2 mg kg-1 ). After one week of germination foliar application of different levels of Zn–lys (0
and 5 ppm) will be given to mediate the detrimental effects of Cd. The seeds will be taken from
Ayub Agricultural Research Institute, Faisalabad (AARI) and will be sown in 64 pots and
seedlings will harvested after 6 weeks of growth to determine various morphological,
physiological and biochemical attributes such as relative membrane stability, H2O2, lipid
peroxidation, MDA contents, antioxidant enzyme activities, free proline accumulation, total
proteins, total free amino acids, total anthocyanin, SOD, POD, ascorbate and CAT at this stages.
Photosynthetic pigments and water contents will be analyzed. Plants will also be analyzed for
lysine, Zn, Cd, Mg, Ca, K etc. The data collected will be subjected to appropriate statistical
analysis.

1
 Government College University, Faisalabad
Department of Botany
Synopsis for MS Degree
Title: Zinc-lysine mediated alterations in morpho-physiological and ionomics
of sunflower (Helianthus annuus L.)under cadmium stress

Date of Admission: 01-06-2017

Date of Initiation: 18-08-2018

Probable Duration: One year

Supervisory committee

Dr. M. Tariq Javed (Supervisor)

Dr. Qasim Ali (Member)

Dr. Naeem Iqbal (Member)

Need of the project

Heavy metals are heterogeneous groups of elements which greatly differ in their chemical
properties and biological functions. Membrane damage, alteration of enzyme activities and the
inhibition of root growth are considered characteristic feature of heavy metal stress (Pourrut et
al., 2013).

Sunflower plant is valuable from economic as well as from an ornamental point of view
sunflower oil is widely used in nutrition and is highly appreciated as a source of the essential
linoleic acid (Wallin et al., 2016)The oil is also used in soap and paints and as a lubricant and the
seeds may be eaten dried or roasted (Dziubanek et al., 2017) The exposure of sunflower
seedlings to 20 mg/kg Cd inhibited biomass production, decreased chlorophyll and carotenoid
concentrations and strongly increased accumulation of Cd in both roots and shoots (Brackhage et
al., 2014).

2
 Cadmium (Cd) is toxic trace pollutant for humans, animals and plants, which enters the
environment mainly from anthropogenic processes and is then transferred to the food chain
(Templeton & Liu., 2010). The health risk associated with cadmium exposure through
consumption of edible plants cultivated in different parts of the world has been estimated when
these plants are eaten by human and animals they cause many disorders. Chronic exposure to
cadmium may result in kidneys and bones damage (Ke at al., 2015).

The negative features of cadmium also include its high mobility within the environment
and bioavailability in the soil-plant system (Shahid et al., 2017). Plants affected by Cd show
impaired photosynthesis (Andosch et al., 2012 ).Cadmium occurs naturally in soils (e.g. in
complexes) but the anthropogenic emissions mostly due to mining activities , burning of fossil
fuels, metallurgical industry and the intensive use of phosphate fertilizers are the main sources of
soil contamination (Singh & Agrawal., 2007).

Cadmium stress in plants leads to a battery of stress symptoms that include chlorosis,
necrotic lesions wilting, disturbances in mineral nutrition and carbohydrate metabolism, and may
therefore strongly reduce biomass production (Santos et al. 2010).

The photosynthetic apparatus is particularly susceptible to Cd and a reduction of

photosynthesis is a common response in plants exposed to Cd (Duan et al., 2013). Cd interacts
with the water balance (Ronzan et al., 2018) and damages the photosynthetic apparatus, in
particular the light harvesting complex II and the photosystem II and I (Goix et al., 2014).
Furthermore, Cd inhibited the oxidative mitochondrial inner membrane (Zanella et al., 2016).

Cd significantly reduces the normal hydrogen and potassium exchange and the activity of
plasma membrane ATPase (Ahmad et al., 2015), and strongly affects (often by inhibiting) the
activity of several enzymes, such as glucose-6-phosphate dehydrogenase, glutamate
dehydrogenase, malic enzyme, isocitrate dehydrogenase .Cadmium toxicity may result from the
binding of metal to sulphydryl groups in proteins, leading to inhibition of activity or disruption
of structure (Ali et al., 2014).

Translocation of cadmium from root to shoot has been studied in several plant species,
showing that it is likely to occur via the xylem and to be driven by leaf transpiration (Khan et al.,
2007). Excess concentrations of Cd may trigger an increased production of the reactive oxygen

3
species (ROS), which includes superoxide anion, hydroxyl radical and hydrogen peroxides
(H2O2) causing membrane damage and electrolyte leakage (Mobin & Khan., 2007).

Cadmium is believed to penetrate the root through the cortical tissue. As soon as Cd
enters the roots, it can reach the xylem through an apoplastic pathway (Howladar, 2015).
Normally Cd ions are mainly retained in the roots, and only small amounts are transported to the
shoots (Singh et al., 2015). It has recently been hypothesized that Cd accumulation in developing
fruits could occur via phloem-mediated transport (Li et al., 2015). Cadmium mainly enters into
the human body by the use of Cd contaminated food crops grown on these contaminated soils
(Nagajyoti el at., 2010).

Zinc-lys significantly increased the photosynthesis, grain yield, enzyme activities and Zn
contents in different plant tissues. Zinc-lys reduced Cd contents in grains, shoot and root as well
as reduced the oxidative stress (Rizwan et al., 2017). Foliar applied Zn-lys reduced HRI (Health
risk index) which may help to reduce health risks associated with Cd.

Hypothesis

 How zinc lysine can alleviate the effect of Cd & mediate the morphological and
physiological attributes of sunflower
 Examination of cd induce alterations in soil system and determine their correlation with
ion dynamics in sunflower plant

Objectives

 To determine how physiological attributes and ion dynamics has changed in sunflower
plant under cadmium stres.
 To study the role of zn lysine in mitigating the cd toxicity in sunfllower

4
Review of Literature

Hendrix et al. (2018) conducted a study to determine the effect of Cd exposure on cell
division and endoreduplication in Arabidopsis thaliana leaves. The result demonstrates that Cd
exposure significantly reduces cell number, cell size and nuclear DNA content, implying an
inhibition of both cell division and endoreduplication. They also examine that these effects
accumulate over time and contribute to Cd-induce disturbance of leaf growth and development.

Garcia et al. (2018) conducted a study to determine the effect of Cd and arsenic on lateral
root formation and development in rice. They reported that Cd and arsenic mainly accumulated
in rice root. The result demonstrates that these heavy metals alter the lateral root primordial
organization and development with negative consequences on root system and affect the cyto-
histology and morphology of the root. This is due to the disturbance of IAA biosynthesis and
transport. Cadmium act on different IAA biosynthetic genes in rice root.

Benavides et al. (2018) studied the impact of Cd and Cu on sunflower plant growth. They
reported that if sunflower plant is treated with 1mM Cdcl2 or Cucl2 for 5-15 days during
germination and seedling it alters the membrane fluidity (MF) of leaves and root. Their results
demonstrate that Cd and Cu have noxious effect on biophysical of plant membrane that could
responsible for toxicity. They concluded that if plant is pre-treated by polyamine or spermine, it
will prevent those alterations.

Yousefi et al. (2018) conduct an experiment in order to evaluate the morphometric

antioxidant enzyme activity and gene expression in sugarcane by growing it in vitro condition
under Cd stress. Sample was cultured in MS medium for 14 days containing CdCl2 (100, 250,
500 µmol). Pigment quantity, antioxidant enzyme activity and gene expression were analyzed.
The amount of Chla, Chlb decrease whereas carotenoid increases significantly. Antioxidant
enzyme activity increase rapidly. They reported that Cd decrease the photosynthetic mechanism
and growth rate in sugarcane. Antioxidant enzyme and gene expression were significantly
unregulated in sugarcane root compare to leaves. Cadmium concentration in shoots and roots of
sugarcane significantly increase.

Hussain et al. (2018) conduct a study to determine the effect of different levels of Zn
lysine (0, 10, 20 and 30 mgL-1) upon photosynthesis, biomass, antioxidant enzyme activity,
5
oxidative stress on rice under various concentrations and Cr. They concluded that Zn-lys foliar
treatment enhance the photosynthesis, biomass, zinc contents and enzyme activity. Their results
reveal that Zn-lys foliar treatment was helpful in increasing plant growth and reduce oxidative
stress also reduce the Cr contents in rice.

Pessoa et al. (2017) investigate the bioimaging of Cd and micronutrients (Cu, Fe, Mn) in
seed of sunflower which grown into Cd contaminated soil. They use laser ablation inductively
plasma mass spectrometer for quantitative approach. Their results indicate that high
concentration of Cd is translocated into seeds and cotyledons ranging from 10 to 20 mg/g-1.
They reported that Cd uptake is responsible for misbalance of micronutrients which play
important role in the metabolism of sunflower. High Cd concentration also causes the failure of
self defense mechanism for avoiding the excess of such elements in aerial compartments of
plant.

Rizwan et al. (2017) studied role of foliar application of zinc-lysine complex on

biochemical, growth parameters and Cd uptake in wheat plant grown under Cd-contaminated
soil. They used different foliar concentrations of zinc-lysine (0, 10, 20 and 30 mgL-1) at different
time intervals (2nd, 3rd, 5th and 7th week). Plant was harvested at maturity. They reported that zinc-
lysine significantly increases photosynthesis, grain yield, enzyme activity, and Zn contents in
different plant tissues. They also examine that Zinc-lysine efficiently improve wheat growth and
fortified Zn contents while reduce Cd concentration in plants. Foliar applied Zinc-lysine reduces
HRI (health risk index) which may reduce the health risks associated with Cd.

Junior et al. (2016) performed in vitro experiment to access the Cd bioavailability and
bioaccessibility. They also evaluate the effect of different doses of Cd (0, 50, 350 and 700 mg)
on sunflower seed. Their results indicate that Cd is translocated to seeds in such that the level of
this element in the seed was 130 times higher than the tolerable amount of human body (0.1 mg
kg-1). While 92%, 72% and 74% of Cd is bioaccessible in control group. They also detect an
imbalance of other essential metals. They reported that Cd interfere with the seed germination by
increasing the dormancy time (50% higher) and by decreasing the germination rate (60% lower).

Zouari et al. (2016) studied the effect of Cd stress on growth and physiological traits in
young date palm and to evaluate the role of exogenous proline in Cd stress alleviation. Two-year

6
old date palm plants were exposed to different Cd concentrations (10 and 30 mg kg-1 soil) for
five months. The result demonstrate that Cd toxicity cause a decrease in growth, mineral
contents, membrane stability, starch contents and peroxidase (POD) activity and an increase in
hydrogen peroxidase concentration. They also reported some physiological symptoms such as
necrosis.

Cheng et al. (2016) conducted an experiment to investigate Cd toxicity absorbent in

aqueous solution. They selected 22 absorbents but find peanut husk biochar as the best absorbent
among them. They reported that husk biochar absorbent approach to equilibrium in 12 hours.
They treated soil with initial 200 mg/L Cd to 40 g/L at room temperature. Cd is adsorbed best at
5 pH within 24 hours by peanut husk biochar within the optimum conditions. Moreover, they
found that Cd ejection reached to 99.9% at these conditions. They concluded that PBH is best
remediate to remove Cd from the polluted environment.

Junior et al. (2015) designed an experiment at proteomic level to evaluate the

biochemical bases of Cd toxicity mechanism in sunflower metabolism. They analyze the changes
in protein abundance in sunflower leaves in the absence or presence of Cd (as CdCl2). They
extract protein with phenol after cultivation period (45 days). The protein content decrease by
approximately 41% in plants treats with 700 mg Cd. They analyze 11 different proteins
identified which involve in energy and disease/defense (including stress), such as ribulose
bisphosphate carboxylase large chain and heat shock proteins were most affected by Cd
exposure. They also concluded that photosynthesis is the main process affected by the presence
of high Cd concentrations in sunflower, although the plants are highly tolerant to Cd stress.

Vaculik et al. (2015) investigated the effect of Si in reducing Cd toxicity in maize. They
performed an experiment by giving two different quantities of Cd and measuring the chloroplast
function in plant cell by using bright field and electron microscopy transmission. They found that
Cd did not influence the leaf anatomy and ultrastructure of mesophyll’s cell of leave. Silicon
showed the negative effect of various biotic and abiotic stresses on plant growth. They found that
Si reduce the syndromes of Cd by increasing effective photochemical quantum of photochemical
system II and content of metabolism of pigments, did not decrease concentration of Cd in leaf
tissues and chloroplast but negatively affect thylakoid formation in chloroplasts of bundle sheath
cells and this was due to alleviated Si. Si induced increased photosynthetic rate and biomass

7
production in C4 plants maize. They concluded that Si reduces the toxicity of Cd by increasing
photosynthetic rate and improve ultrastructure in chloroplasts of bundle sheath’s cell.

Rashti et al. (2014) reported that to forecast environmental issue it is essential to

understand the desorption nature of pollutants in soil system. They performed an experiment to
study the soil behavior for Cd absorption. Their treatments consisted of Cd in two levels
(15mg/kg and 30 mg/kg as CdCl2). Soil were incubated at 25± 1 C and 20%. Cd absorption
pattern was examined by DTPA extraction from 30 to 960min. Result showed that constant rate
of kinetic models exhibited best correlation with each other. They concluded that pH, CEC, Fe
and OM are essential characters of soil that affected Cd desorption and diffusion.

Klaus et al. (2013) designed a study to investigate the effect of cadmium on

photosynthetic pigments and growth of maize plants at different stages. At germination stage
they used 2mM concentration of Cd, at seedling stage they used 4-200 µM (4-9 days) and
juvenile plants (34 days). They demonstrated that the high amount of Cd stored in roots while
only small amount translocated to maize shoots. They declared that the high amount of Cd does
not show any effect on germinating stage while the seedling stage is most sensitive stage it
shows effect on seedling stage, and 90% growth inhibition takes place in juvenile stage. They
also exhibited that the Cd influenced the shoot mass accumulation more than that of root mass.
They additionally exhibited that in juvenile stage shoot water substance is more influenced than
root. They inferred that the affectability of Cd expanded with expanding age of the plants and
shoot development is more influenced by Cd than roots.

Hussain et al. (2013) studied the inhibitory effect of different concentration of Cd on growth and
biochemical parameters in two maize varieties at various time intervals. They applied different
levels (0, 3, 6,9mg/kg) of CdCl2 on 20 days old two maize lines i.e. Maize- targeted mutagenesis
1and 2 in sand. They got results after 5th, 10th, 15th days of treatment and considerably reduced
plant growth in line maize-targeted mutagenesis-1 while plant biomass and leaf area had no
inhibitory effects due to Cd in line maize-targeted mutagenesis-2 at 3mg of CdCl2 /kg. Results
suggested that H2O2, Malondialdehyde contents, Relative membrane permeability and
anthocyanin increased at all level of Cd in both lines while growth, photosynthetic pigment, free
amino acid and proline contents was reduced due to induced oxidative stress specially at 10th
and 15th day after treatments. They concluded that Cd affect morph physiological parameters in

8
both lines maize-targeted mutagenesis while maize-targeted mutagenesis was considered as more
tolerant than maize-targeted mutagenesis.

Irfan et al. (2013) explained effects of Cd toxic soil and its physiological damages
through their experimental study. They reported that Cd toxicity induced nutritional stress with
physiological and toxic symptoms. Cadmium also induced oxidative stress which stops defense
mechanism and accelerates damages at the expense of growth. Through mineral uptake system
many undesirable elements were taken along with Cd. Different pathways were associated with
cellular sequestration and systematic accumulation of Cd. They also reported reduced carbon
fixation and photosynthesis, electron leakage as ROS, enhanced negative feedback in plants.
Taken together results showed that Cd caused loss of crop yield.

Nazar et al. (2012) conducted an experiment to investigate the nutrients uptake under Cd
stress conditions in plants. They reported that Cd and nutrients share the same transport channels
for accumulation into roots, so due to increase in Cd uptake, the supply of nutrients in plants will
decrease. They also reported that increase into the concentrations of nutrients is beneficial in
influence the Cd stress and it may outcomes in the increase in plant growth and accumulation of
biomass.

Hussain et al. (2012) explored the impact of Cd on two maize cultivars. They conducted
the pot experiment in growth chamber after15 days of germination. They treated the sand with
half quality Hoagland's supplement arrangement having 0, 25, 50 and 75 μM kg-1 Cd fixations.
They reported that Cd in high concentration extremely diminished the different growth
parameters in and no impact reported up to the 50 μM Cd. They reported that Cd reduces the
relative water content, and chlorophyll while enhanced membrane permeability, hydrogen
peroxide and malondialdehyde at all levels. They also reported that the Agatti-2002 is a Cd
tolerant. They concluded that to overcome Cd toxicity cultivate Cd tolerant varieties.

Kostandi et al. (2012) investigated impact of Cd on the uptake of nutrients and

rhizosphere pH of two maize hybrids (yellow and white). They conducted experiment under
greenhouse effect and used the soil having pH 8. They applied Cd concentration that is (0, 2, 5
mg/kg) and two nitrogen fertilizers that is NH4+ and NO3-. They demonstrated that yellow
cultivar has high shoot dry weight and low uptake of Cd than the white cultivar. They also

9
demonstrated that the accumulation of the nitrogen in plants was not affected by the form of
nitrogen that is present in both cultivars. They also demonstrated that the rhizospheric pH of soil
reduced in both forms of nitrogen that is present in both cultivars. N level is high in soil in NH4+
and low in NO3- after harvest.

Materials and Methods

The present research conduct at research area, Government College University,

Faisalabad, Pakistan to investigate the Zn lysine mediated alterations on physiology and yield of
sunflower under Cd stress.

Seed Material

Seed of sunflower hybrid variety will be use in this experiment. The seeds will be obtained from
Ayyub Research Institute Faisalabad, Pakistan.

Procedure

Soil obtained from Faisalabad will be filled in 64 pots (8 kg per Pot). Seeds of sunflower
will be sown in each pot. After germination Cd solution of different concentrations will be added
in each pot. After week 1 zinc lysine will be foliarly applied. After week 4 harvesting will be
done from each pot. Three plants will be collected from each pot. The fresh weight of one plant
will be taken and the plant will be stored in paper bag. Two fresh plants will be taken in zipper
bags. Data will be collected according to completely randomized design with four replications.
Remaining plants will be examined to reach maturity.

Data Collection

Following morphological, biochemical and physiological parameters will be recorded after

harvesting.

Morphological analysis

 Plant height (cm)

 Shoot fresh weight (g)
 Shoot dry weight (g)

10
 Root fresh weight (g)
 Root dry weight (g)
 Leaf fresh weight (g)
 Leaf dry weight (g)
 Leaf area (cm2)
 Leaf relative water contents

Biochemical analysis
 Ascorbic acid concentration
 H2O2 contents
 MDA contents
 Total proline
 Catalase activity
 Peroxidise
 Superoxide dismutase

Physiological analysis
 Determination of nutrients
 Total phenolics
 Total free amino acids
 Total soluble protein contents
 Total flavonoids
 Total soluble sugars
 Chlorophyll and carotenoids contents

Ionomic analysis
 Lys-
 Zn+
 Mg+
 Ca+

11
 K+

Statistical Analysis

The data collected will be analyzed by using variance (ANOVA) with completely randomized
design (CRD) with four replicates per treatment.

References

Adams, S. V., Quraishi, S. M., Shafer, M. M., Passarelli, M. N., Freney, E. P., Chlebowski, R.
T., ... & Newcomb, P. A. (2014). Dietary cadmium exposure and risk of breast,
endometrial, and ovarian cancer in the Women’s Health Initiative. Environmental Health
Perspectives, 122(6), 594.
Ahmad, P., Sarwat, M., Bhat, N. A., Wani, M. R., Kazi, A. G., & Tran, L. S. P. (2015).
Alleviation of cadmium toxicity in Brassica juncea L.(Czern. & Coss.) by calcium
application involves various physiological and biochemical strategies. PloS one, 10(1),
e0114571.
Ali, B., Gill, R. A., Yang, S., Gill, M. B., Ali, S., Rafiq, M. T., & Zhou, W. (2014). Hydrogen
sulfide alleviates cadmium-induced morpho-physiological and ultrastructural changes in
Brassica napus. Ecotoxicology and Environmental Safety, 110, 197-207.
Andosch, A., Affenzeller, M. J., Lütz, C., & Lütz-Meindl, U. (2012). A freshwater green alga
under cadmium stress: ameliorating calcium effects on ultrastructure and photosynthesis
in the unicellular model Micrasterias. Journal of Plant Physiology, 169(15), 1489-1500.
Azevedo, H., Glória Pinto, C. G., & Santos, C. (2005). Cadmium effects in sunflower: membrane
permeability and changes in catalase and peroxidase activity in leaves and
calluses. Journal of Plant Nutrition, 28(12), 2233-2241.
Benavides, M. P., Gallego, S. M., & Tomaro, M. L. (2005). Cadmium toxicity in
plants. Brazilian Journal of Plant Physiology, 17(1), 21-34.
Benavides, M. P., Groppa, M. D., Recalde, L., & Verstraeten, S. V. (2018). Effects of
polyamines on cadmium-and copper-mediated alterations in wheat (Triticum aestivum
L) and sunflower (Helianthus annuus L) seedling membrane fluidity. Archives of
Biochemistry and Biophysics.

12
Brackhage, C., Huang, J. H., Schaller, J., Elzinga, E. J., & Dudel, E. G. (2014). Readily available
phosphorous and nitrogen counteract for arsenic uptake and distribution in wheat
(Triticum aestivum L.). Scientific Reports, 4, 4944.
Cataldo, D. A., McFadden, K. M., Garland, T. R., & Wildung, R. E. (1988). Organic constituents
and complexation of nickel (II), iron (III), cadmium (II), and plutonium (IV) in soybean
xylem exudates. Plant Physiology, 86(3), 734-739.
Cheng, Q., Huang, Q., Khan, S., Liu, Y., Liao, Z., Li, G., & Ok, Y. S. (2016). Adsorption of Cd
by peanut husks and peanut husk biochar from aqueous solutions. Ecological
Engineering, 87, 240-245.

Dias, M. C., Monteiro, C., Moutinho-Pereira, J., Correia, C., Gonçalves, B., & Santos, C. (2013).
Cadmium toxicity affects photosynthesis and plant growth at different levels. Acta
Physiologiae Plantarum, 35(4), 1281-1289.
Duan, G., Liu, W., Chen, X., Hu, Y., & Zhu, Y. (2013). Association of arsenic with nutrient
elements in rice plants. Metallomics, 5(7), 784-792.
Dziubanek, G., Baranowska, R., Ćwieląg-Drabek, M., Spychała, A., Piekut, A., Rusin, M., &
Hajok, I. (2017). Cadmium in edible plants from Silesia, Poland, and its implications for
health risk in populations. Ecotoxicology and Environmental Safety, 142, 8-13.
Goix, S., Lévêque, T., Xiong, T. T., Schreck, E., Baeza-Squiban, A., Geret, F., ... & Dumat, C.
(2014). Environmental and health impacts of fine and ultrafine metallic particles:
assessment of threat scores. Environmental Research, 133, 185-194.
Howladar, S. M. (2014). A novel Moringa oleifera leaf extract can mitigate the stress effects of
salinity and cadmium in bean (Phaseolus vulgaris L.) plants. Ecotoxicology and
Environmental Safety, 100, 69-75.
Hussain, A., Ali, S., Rizwan, M., ur Rehman, M. Z., Hameed, A., Hafeez, F., ... & Wijaya, L.
(2018). Role of Zinc–Lysine on Growth and Chromium Uptake in Rice Plants under Cr
Stress. Journal of Plant Growth Regulation, 1-10.
Hussain, I., Akhtar, S., Ashraf, M. A., Rasheed, R., Siddiqi, E. H., & Ibrahim, M. (2013).
Response of maize seedlings to cadmium application after different time intervals. ISRN
Agronomy, 2013.
Hussain, I., Iqbal, M., QURAT-UL-AIN, S. O. B. I. A., Rasheed, R., Mahmood, S., Perveen, A.,
& Wahid, A. (2012). Cadmium dose and exposure-time dependent alterations in growth

13
 and physiology of maize (Zea mays). International Journal of Agriculture and
Biology, 14(6).
Irfan, M., Hayat, S., Ahmad, A., & Alyemeni, M. N. (2013). Soil cadmium enrichment:
Allocation and plant physiological manifestations. Saudi Journal of Biological
Sciences, 20(1), 1-10.

Júnior, C. A. L., de Sousa Barbosa, H., Galazzi, R. M., Koolen, H. H. F., Gozzo, F. C., &
Arruda, M. A. Z. (2015). Evaluation of proteome alterations induced by cadmium stress
in sunflower (Helianthus annuus L.) cultures. Ecotoxicology and Environmental
Safety, 119, 170-177.
Júnior, C. A. L., Oliveira, S. R., Mazzafera, P., & Arruda, M. A. Z. (2016). Expanding the
information about the influence of cadmium on the metabolism of sunflowers:
Evaluation of total, bioavailable, and bioaccessible content and metallobiomolecules in
sunflower seeds. Environmental and Experimental Botany, 125, 87-97.
Ke, S., Cheng, X. Y., Zhang, J. Y., Jia, W. J., Li, H., Luo, H. F., ... & Chen, Z. N. (2015).
Estimation of the benchmark dose of urinary cadmium as the reference level for renal
dysfunction: a large sample study in five cadmium polluted areas in China. BMC Public
Health, 15(1), 656.
Khan, N. A., Samiullah, Singh, S., & Nazar, R. (2007). Activities of antioxidative enzymes,
sulphur assimilation, photosynthetic activity and growth of wheat (Triticum aestivum)
cultivars differing in yield potential under cadmium stress. Journal of Agronomy and
Crop Science, 193(6), 435-444.
Kostandi, S. F., Soliman, M. F., Beschow, H., & Merbach, W. (2012). Rhizosphere effects of
maize hybrids and N forms on Cd bioavailability in a limed soil. Archives of Agronomy
and Soil Science, 58(8), 903-913.

Mobin, M., & Khan, N. A. (2007). Photosynthetic activity, pigment composition and
antioxidative response of two mustard (Brassica juncea) cultivars differing in
photosynthetic capacity subjected to cadmium stress. Journal of Plant
Physiology, 164(5), 601-610.
Nagajyoti, P. C., Lee, K. D., & Sreekanth, T. V. M. (2010). Heavy metals, occurrence and
toxicity for plants: a review. Environmental Chemistry Letters, 8(3), 199-216.

14
Nazar, R., Iqbal, N., Masood, A., Khan, M. I. R., Syeed, S., & Khan, N. A. (2012). Cadmium
toxicity in plants and role of mineral nutrients in its alleviation. American Journal of
Plant Sciences, 3(10), 1476.

Pant, N., Kumar, G., Upadhyay, A. D., Gupta, Y. K., & Chaturvedi, P. K. (2015). Correlation
between lead and cadmium concentration and semen quality. Andrologia, 47(8), 887-
891.
Pessôa, G. D. S., Júnior, C. A. L., Madrid, K. C., & Arruda, M. A. (2017). A quantitative
approach for Cd, Cu, Fe and Mn through laser ablation imaging for evaluating the
translocation and accumulation of metals in sunflower seeds. Talanta, 167, 317-324.
Pourrut, B., Shahid, M., Douay, F., Dumat, C., & Pinelli, E. (2013). Molecular mechanisms
involved in lead uptake, toxicity and detoxification in higher plants. In Heavy Metal
Stress in Plants (pp. 121-147). Springer, Berlin, Heidelberg.
Rashti, M. R., Esfandbod, M., Adhami, E., & Srivastava, P. (2014). Cadmium desorption
behaviour in selected sub-tropical soils: Effects of soil properties. Journal of
Geochemical Exploration, 144, 230-236.

Rizwan, M., Ali, S., Hussain, A., Ali, Q., Shakoor, M. B., Zia-ur-Rehman, M., ... & Asma, M.
(2017). Effect of zinc-lysine on growth, yield and cadmium uptake in wheat (Triticum
aestivum L.) and health risk assessment. Chemosphere, 187, 35-42.
Ronzan, M., Zanella, L., Fattorini, L., Della Rovere, F., Urgast, D., Cantamessa, S., ... &
Feldmann, J. (2017). The morphogenic responses and phytochelatin complexes induced
by arsenic in Pteris vittata change in the presence of cadmium. Environmental and
Experimental botany, 133, 176-187.
Saidi, I., Chtourou, Y., & Djebali, W. (2014). Selenium alleviates cadmium toxicity by
preventing oxidative stress in sunflower (Helianthus annuus) seedlings. Journal of Plant
Physiology, 171(5), 85-91.
Shahid, M., Dumat, C., Khalid, S., Niazi, N. K., & Antunes, P. M. (2016). Cadmium
bioavailability, uptake, toxicity and detoxification in soil-plant system. In Reviews of
Environmental Contamination and Toxicology Volume 241 (pp. 73-137). Springer,
Cham.

15
Singh, R. P., & Agrawal, M. (2007). Effects of sewage Helianthus annuus L) sludge amendment
on heavy metal accumulation and consequent responses of Beta vulgaris
plants. Chemosphere, 67(11), 2229-2240.
Templeton, D. M., & Liu, Y. (2010). Multiple roles of cadmium in cell death and
survival. Chemico-Biological Interactions, 188(2), 267-275.
Vaculík, M., Pavlovič, A., & Lux, A. (2015). Silicon alleviates cadmium toxicity by enhanced
photosynthetic rate and modified bundle sheath's cell chloroplasts ultrastructure in
maize. Ecotoxicology and Environmental Safety, 120, 66-73.

Yousefi, Z., Kolahi, M., Majd, A., & Jonoubi, P. (2018). Effect of cadmium on morphometric
traits, antioxidant enzyme activity and phytochelatin synthase gene expression (SoPCS)
of Saccharum officinarum var. cp48-103 in vitro. Ecotoxicology and Environmental
Safety, 157, 472-481.
Zanella, L., Fattorini, L., Brunetti, P., Roccotiello, E., Cornara, L., D’Angeli, S., ... & Degola, F.
(2016). Overexpression of AtPCS1 in tobacco increases arsenic and arsenic plus
cadmium accumulation and detoxification. Planta, 243(3), 605-622.
Zouari, M., Elloumi, N., Ahmed, C. B., Delmail, D., Rouina, B. B., Abdallah, F. B., &
Labrousse, P. (2016). Exogenous proline enhances growth, mineral uptake, antioxidant
defense, and reduces cadmium-induced oxidative damage in young date palm (Phoenix
dactylifera L.). Ecological Engineering, 86, 202-209.

16
 Signatures
Name of Student: Farwa jabeen ___________________

Supervisory Committee (Name and Signature)

1) Dr. M. Tariq Javed (Supervisor) __________________

2) Dr. Qasim Ali (Member) __________________

3) Dr. Naeem Iqbal (Member) __________________

Faculty Scrutiny committee (Name and Signature)

1) Prof. Dr. Muhammad Iqbal _____________________

2) Dr. Nudrat Aisha Akram _____________________

3) Dr. Muhammad Tariq Javed _____________________

4) Dr. Shgufta Perveen _____________________

___________________________ _________________

Chairman Dean

Department of Botany Faculty of Life Sciences

____________________________

Director of Advanced Studies

17