Kaur Nayyar : Heavy metal toxicity to food legumes: effects, antioxidative defense and tolerance mechanisms 1 1
commonly found in leaves, floral parts, and pollens. They usually accumulate in the plant vacuole as glycosides, but
they also occur as exudates on the surface of leaves and other aerial plant parts. Flavonoids are suggested to have
many functions in flowers, fruits, and seed pigmentation, protection against UV light, defence against phytopathogens
pathogenic microorganisms, insects, animals, role in plant fertility and germination of pollen and, molecules in plant-
microbe interaction. Apart from the above roles, flavonoids have as antioxidative activity Brown et al.1998. Besides
having the function of ROS scavenging , flavonoids are able to function as chelators for metals, depending on the molecular
structure Brown et al.1998 and hence can take part in plant defence. In Arabidopsis thaliana, the relation between
flavonoids and heavy metal tolerance were investigated. Both Arabidopsis
wild type and mutant lines with a defect in flavonoid biosynthesis were grown on media containing
different heavy metals. Results revealed that root length and seedling weight were reduced in mutants more than in the
wild type when grown on cadmium, while on zinc only root length was affected Keilig and Muller 2009.
5. Tolerance mechanisms
Heavy metals in the plant environment operate as stress factors that cause physiological strain and in doing so they
reduce the plant vigour and totally inhibit the plant growth in extremes However, plants have evolved several physiological
mechanisms which enable them to tolerate metal toxicity Baker 1987. The development of metal tolerance in plants is a major
way to reduce the harmful effects of excessive exposure to heavy metal ions Tyler et al.1989. There are various potential
cellular and other mechanisms available for metal detoxification and tolerance in higher plants Hall 2002, which
have been reported to function in legumes.
5.a. Role of Arbuscular mycorrhizal fungi
Mycorrhizal association is a symbiotic non-pathogenic relationship between plant roots and fungal hyphae with a
fungal connection between the soil and the root Harley and Smith 1983. It has been reported that the host plant receives
support from AM fungi, with the help of its symbiotic association, in the aspect of uptake of phosphorus and other
nutrients, enhancement of growth hormones, increase of protein content, increase of lipid, sugars, amino acid levels,
increase of tolerance to heavy metals, increase of salinity tolerance, and resistance to root-borne pathogens
Upadhyaya et al. 2010. Recently, the symbiotic association with mycorrhizal fungi has been proposed as one of the major
mechanisms of plant HM-tolerance Hall 2002, Joachim et al.
2009. However, alleviating heavy metal toxicity by AMF colonization can vary to a large extent, depending on which
heavy metal is involved, its concentration in the soil, the fungal symbiosis partner and the conditions of plant growth Turnau
1998. There are many
strategies adopted by AM which can
alleviate heavy metal threats in mixed
culture systems and, thus, from the food
chains Joschim et al. 2009. These include
the immobilisation of metal compounds,
precipitation
of p o l y p h o s p h a t e
granules in the soil, adsorption to chitin in
the fungal cell walls and chelation of
heavy metals inside the fungus Joachim et al. 2009. Generally, AM binds to heavy metals beyond the plant rhizosphere by
releasing an insoluble glycoprotein commonly known as glomalin Gonzalez- Chavez et al. 2004. The roles of AM are
summarized in the Fig.5.
Numerous studies have indicated that AMF can decrease the metal uptake of the host plants, thus protecting
them against HMs toxicity Leyval et al. 1997. Many heavy metal contaminated sites are reported to have mycorrhizae
Weissenhorn and Leyval 1993.This indicates that these fungi have evolved a HM-tolerance and that they may play a role in
the phytoremediation of the site Khan et al. 2000. Mycorrhizae were found to ameliorate the toxicity of trace
metals in polluted soils growing in soybean and lentil plants Jamal et al. 2002. Increased heavy metal tolerance of plants
by dual inoculation of an arbuscular mycorrhizal fungi and nitrogen-fixer Rhizobium bacterium was reported in cowpea
Al-Garni 2006. The effects of dual inoculation with arbuscular mycorrhizal AM fungus and Rhizobium N-fixing bacteria,
NFB on the host plant cowpea Vigna sinensis in pot cultures were investigated at six concentrations of Zn 0.0-1000 mgkg
dry soil and Cd 0.0-100 mgkg dry soil. The study provides evidence for benefits of NFB to AM fungi in the protection of
host plants against the detrimental effects of heavy metals and provides the mechanisms for metal tolerance against
them.A greenhouse pot experiment was done to investigate the effects of the colonization of arbuscular mycorrhizal fungus
AMF Glomus mosseae on the growth and metal uptake of three leguminous plants Sesbania rostrata, Sesbania
cannabina
, Medicago sativa grown in multi-metal contaminated soil Lin et al. 2007. The results revealed that
AMF colonization increased the growth of the legumes thereby indicating that AMF colonization increased the plant’s
resistance to heavy metals. The effect was also enhanced on the formation of root nodules and N and P uptake increased,
which may be due to the heavy metal tolerance mechanisms conferred by the AMF.
Fig: 5 Steps for alleviating heavy metal stress adopted by AM fungi
1 2 Journal of Food Legumes 263 4, 2013
5.b. Role of metallothioneins and phytochelatins
Metallothioneins MT’s belong to a family of cysteine- rich low molecular weight metal-binding proteins generally
induced during the metal stress Corbett and Goldsbrough 2002. Metallothioneins generally form complexes with heavy
metal ions and are present in almost all forms of life and have a role in protecting cells from the deleterious effects of high
concentration metal ions. The function of MT is to detoxify non-essential metals such as mercury, cadmium and essential
metals such as zinc and copper.
Phytochelatins PCs, a type of MT’s, are synthesized in plants in response to heavy metal stress and due to various
metals. PCs consist of only the three amino acids: Glu, Cys and Gly, the Glu, and Cys residues linked through a g-
carboxylamide bondCobbet 2000.Recent research indicates that PCs are present in a wide variety of plant species and in
some microorganisms. They are structurally related to glutathione GSH, g-Glu-Cys-Gly and were presumed to be
the products of a biosynthetic pathway. In addition, a number of structural variants, for example, g-Glu-Cysn-b- Ala, g-
Glu-Cysn-Ser, and g-Glu-Cysn-Glu, have been identified in some plant species Rauser 1999, Zenk 1996.
Activation of the detoxicative-phytochelatin system was observed in the cytosol of root cells of three legume species,
Vicia faba , Pisum sativum, and Phaseolus vulgaris when they
were exposed to lead ions Piechalak et al. 2002. This system was composed of phytochelatins PCs in roots of V. faba,
homophytochelatins hPCs in P. vulgaris roots, and both PCs and hPCs in P. sativum roots.
5.c. Organic acids and amino acids
Some amino acids, particularly histidine and proline, also play very important roles in the chelation of metal ions
both within plant cells and in the xylem sap Rai 2002. Kerkeb and Krämer 2003have reported that in Alyssum lesbiacum
and Brassica juncea , an enhanced release of Ni into the xylem is associated with concurrent release of histidine from an
increased root free His pool. Other amino acids such as citrate, malate and histidine are potential ligands for heavy metals
and could play a role in tolerance and detoxification Rauser 1999. Citrate, malate and oxalate have been involved in
transport of metal ions through the xylem and vacuolar sequestering Rauser 1999. It is reported that citric acid to be
a major Cd
2+
ligand at low Cd
2+
concentrations Wagner 1993 and has been shown to form complexes with Ni
2+
in Ni- hyperaccumulation plants Sagner et al. 1998. It is also
suggested that malate is a cytosolic zinc chelator in zinc- tolerant plants Mathys 1977. Kramer et al.1996 reported
that the significant and proportional change in amino acid or organic acid concentration elicited by a change in metal
exposure was shown by histidine response in plants that accumulate nickel. The presence of different concentrations
of organic acids among various ecotypes of metal-tolerant plants in their natural habitat has deemed these substances
as likely cellular chelators Rauser 1999.
5.d. Polyamines
Polyamines PAs are nitrogenous compounds present in all living cells. They are not only involved in various cellular
processes like growth promotion and cell division but also in the inhibition of ethylene production and senescence Tiburcio
et al. 1997. They influence a variety of growth and
development processes in plants which have been suggested to be a class of plant growth regulators and to act as second
messengers Evans and Malmberg 1989, Kakkar and Sawhney 2002. The polyamines are cations due to protonation at
cytoplasmic pH, i.e. putrescine
2+
, spermidine
3+
, and spermine
4+
, which accounts for their binding ability to nucleic acids Flink
and Pettijohn 1975.It has been reported that the levels of polyamines and the activities of their biosynthetic enzymes
in plants increase under environmental stresses Evans and Malmberg 1989. Polyamine contents are highly altered in
response to the exposure to heavy metals. For example, the response of different polyamines to Cd treatment strongly
varied in Phaseolus vulgaris in an organ-specific manner. Putrescine increased in root, hypocotyl, and epicotyl whereas
spermidine increased in hypocotyl, decreased in leaves, and did not change in roots.In soybean phospholipids, using
membrane vesicles Weinstein et al. 1986. Tadolini et al. 1984 showed that polyamines inhibit lipid peroxidation when bound
to the negative charges on the vesicle surface. In addition, polyamines namely, spermine, spermidine, putrescine, and
cadaverine have been demonstrated to scavenge free radicals in vitro
Drolet et al.1986. Furthermore, polyamines block one of the major vacuolar channels, the fast vacuolar cation
channel, and their accumulation could decrease ion conductance at the vacuolar membrane to facilitate metal ion
compartmentation Bru¨ggemann et al.1998. Their roles in metal tolerance remain to be explored in detail.
Conclusion
Heavy metal contaminations seriously threaten the productivity of plants and particularly the legumes which are
important atmospheric nitrogen fixers and an excellent source of protein to both animals and human beings. These metals
prove to be deleterious for the legume growth and physiology, and ultimately enter the food chain to affect human population.
In spite of presence of diverse tolerance mechanisms to toxic metals, legumes suffer due to their cultivation in contaminated
soils. The knowledge about metal tolerance mechanisms gained from model plants such as Arabidopsis needs to be
explored in legumes too to induce tolerance to metals. Various mechanisms involving phytochelatins, thiols, transporters
based in plasma membrane and tonoplast need to be manipulated through genetic means to enhance the metal
tolerance in legumes.
Kaur Nayyar : Heavy metal toxicity to food legumes: effects, antioxidative defense and tolerance mechanisms 1 3
Acknowledgement
The Financial assistance in the form of fellowship from University Grants Commission, New Delhi, to the first author
is gratefully acknowledged.
References
Athar R and Ahmad M. 2002. Heavy metal toxicity in legume –
microsymbiont system. Journal of Plant Nutrition.25:369-386.
Al-Garni SMS. 2006. Increased heavy metal tolerance of cowpea plants by dual inoculation of an arbuscular mycorrhizal fungi and nitrogen
fixer rhizobium. African Journal of Biotechnology.5:133-142.
Ahmad E, Almas Z, Mohammad SK and Mohammad O.2012. Heavy metal Toicity to Symbiotic Nitrogen-Fixing microorganism and
Host Legumes.In Toxicity of heavy metals to legume and bioremediation. Eds.A.Zaidi,PA Wani and MS Khan.Pp 248.
Ahmad MSA, M Hussain, S Ijaz and AK Alvi. 2008. Photosynthetic performance of two mung bean Vigna radiata L. wilczek cultivars
under lead and copper stress. International Journal of Agriculture and Biology.10: 167–72.
Alia and Saradhi PP. 1991. Proline accumulation under heavy metal
stress.Journal of Plant Physiology 138:554-558.
Al-Qurainy Fahad 2009. Toxicity of Heavy Metals and Their Molecular Detection on Phaseolus vulgaris L. Australian Journal of Basic
Applied Sciences.3:3025
Al-Rumaih MM, Rushdy SA and Warsy AS. 2001. Effect of cadmium chloride on seed germination and growth characteristics of cowpea,
Vigna unguiculata L. plants in the presence and absence of gibberellic
acid. Saudi Journal of Biological Sciences 8:41-51.
Al-Yemeni and Mohammed-Nasser. 2001. Effect of cadmium, mercury and lead on seed germination and early seedling growth of Vigna
ambacensis
L. Indian Journal of Plant Physiology 6: 147-151.
Angelov MT, Tsonev A, Uzunova K and Gaidardjieva.1993. Cu
2+
effect upon photosynthesis, chloroplast structure, RNA and protein
synthesis of pea plants. Photosynthetica 28
: 341-350. Antipchuk AF, Rangelova VN and Tantsurenko FV. 2000. Effects of
heavy metals and reclaimerson formation and functioning of legume – rhizobial symbiosis. Mikrobiologi Chnii-Zhurnal 62:44-50.
Arora NK, Khare E, Singh S and Maheshwari DK. 2010.Effect of aluminium and heavy metals on enzymes of nitrogen metabolism
of fast and slow growing rhizobia under explanta conditions. World Journal of Microbiology and Biotechnology 26:811–816.
Atici O, H Outcu and OF Algur. 2005. Effect of putrescine on inducing symbiosis in chickpea and vetch inoculated with commercial or
indigenous strains of Rhizobium. Symbiosis.38: 163-174.
Aydinalp C and Marinova S. 2009. The effects of heavy metals on seed germination and plant growth on alfalfa plant Medicago sativa.
Bulgarian Journal of Agricultural Science15:347-350.
Babich H and Stotzky G. 1978. Effects of cadmium on the biota: Influence of environmental factors. Advances in Applied
Microbiology 23:55–117. Baker AJM.1987. Metal tolerance.New Phytologist .106:93-111.
Bakkaus E, Gouget B, Gallien JP, Khodja H, Carrot H, Morel JL and Collins R. 2005. Concentration and distribution of cobalt in higher
plants: the use of micro-PIXE spectroscopy. Nuclear Instruments and Methods. 231:350–356.
Balestrasse,KB, Gallego SM and Tomarol ML. 2004. Cadmium-induced senescence in nodules of soybean Glycine max L. plants.Plant
and Soil.262: 373–381.
Barceló J and Poschenrieder CM. 1999. Structural and ultrastructural changes in heavy metal exposed plants. In. Heavy Metal Stress in
Plants, from Molecules to Ecosystems. Eds Prasad MNV and Hagemeyer J . Springer Verlag, Berlin.Pp183-205.
Barcelo J, Vazquez MD and Poschenrieder CH. 1987. Structural and ultrastructural disorders in cadmium-treated bush bean plants
Phaseolus vulgaris L.. New Phytologist 108:37–49.
Baszynski T, Wajda L, Krol M, Wolinska D, Krupa Z and Tukendorf A. 1980. Photosynthetic activities of cadmium-treated tomato plants.
Plant Physiology 48:365–370.
Bera AK, Kanta-Bokaria AK and Bokaria K. 1999.Effect of tannery effluent on seed germination, seedling growth and chloroplast
pigment content in Mungbean Vignaradiata LWilczek. Environment and Ecology 17:958–961.
Bhardwaj P, Chaturvedi AK and Prasad P. 2009. Effect of enhanced lead and cadmium in soil on physiological and biochemical attributes
of Phaseolus vulgaris L. Nature and Science7:63-75.
Bhat MT, Ansari MY, Choudhary S, Aslam R and Alka. 2011. Synergistic Cytotoxic Stress and DNA Damage in Clover Trifolium repens
Exposed to Heavy Metal Soil from Automobile Refining Shops in Kashmir-Himalaya. ISRN Toxicology2011:1-7.
Bhamburdekar SB and Chavan PD. 2011. Effect of Some Stresses on Free Proline Content During Pigeonpea Cajanas cajan Seed
Germination. Journal of Stress Physiology Biochemistry.7: 235- 241.
Bianucci E, Fabra A and Castro S. 2011.Cadmium Accumulation and Tolerance in Bradyrhizobium spp. Peanut Microsymbionts.
Current Microbiology 62:96–100.
Bibi M and Hussain M. 2005.Effect of copper and lead on photosynthesis and plant pigments in black gram Vigna mungo L.Hepper.
Bulletin of Environmental Contamination and Toxicology 74
:1126–33. Blaylock MJ and Huang JW. 2000. Phytoextraction of metals.
Phytoremediation of toxic metals: using plants to clean up the environment. Eds. Raskin I and Ensley BD. John Wiley and Sons,
Toronto.Pp303. Böddi B, Oravecz AR and Lehoczki E. 1995.Effect of cadmium on
organization and photoreduction of protochlorophyllide in dark- grown leaves and etioplast inner membrane preparations of wheat.
Photosynthetica 31:411-420.
Bouazizi H, Jouili H, Geitmann A and Ferjani EEI. 2010. Copper toxicity in expanding leaves of Phaseolus vulgaris L.: antioxidant enzyme
response and nutrient element uptake. Ecotoxicology and Environmental Safety 73:1304–1308.
Broos K, Beyens H and Smolders E. 2005. Survival of rhizobia in soil is sensitive to elevated zinc in the absence of the host plant. Soil
Biology and Biochemistry 37:573–579.
Brown JE, Khodor H, Hider RC and Rice-Evans CA. 1998. Structural dependence of flavonoid interactions with Cu2+ ions: implications
for their antioxidant properties. Biochemical Journal330: 1173-1178.
Brüggemann LI, Pottosin I I and Schönknecht G. 1998. Cytoplasmic polyamines block the fast-activating vacuolar cation channel. the
Plant Journal16:101–105.
Brun LA, Maillet J, Hinsinger P and Pepin M. 2001.Evaluation of copper availability to plants in copper-contaminated vineyard soils.
Environmental Pollution 111: 293–302.
1 4 Journal of Food Legumes 263 4, 2013
Brynhildsen L and Rosswall T. 1997. Effects of metals on the microbial
mineralization of organic acids. Water, Air, and Soil Pollution 94:45–57.
Budnikov G. 1998. Heavy metals in the ecological monitoring in water
systems.Educational Journal of Trorosovsk. 5: 23-29 In Russian.
Cobbett C and Godsbrough P. 2002. Phytochelatins and metallothioneins: Roles in heavy metal detoxification and
homeostatis. Annual Review of Plant Biology.53:159-82.
Cobbett CS. 2000. Phytochelatins and heavy metal tolerance in
plants.Current Opinion in Plant Biology.3:211-216.
Cakmak I, Horst WJ and Bangruth F. 1989. Effect of Zinc Nutritional Status on Growth, Protein Metabolism and Levels of Indole-3-
acetic acid and other phytohormones in bean Phaseolus vulgaris L.. Journal of Experimental Botany.40:405-412.
Cakmak I and Horst WJ. 1991. Effect of aluminum on lipid peroxidation, superoxide dismutase, catalase and peroxidase activities in root tips
of soybean Glycine max .Physiolgia Plantarum. 83: 463-468.
Cargnelutti D, Tabaldi LA, Spanevello RM, Jucoski GO, Battisti V, Redin M, Linares CEB, Dressler VL, Flores MM, Nicoloso FT,
Morsch VM and Schetinger MRC. 2006. Mercury toxicity induces oxidative stress in growing cucumber seedlings. Chemosphere
65
:999–1006. Cervantes C, Campos-Garcia J, Debars S, Gutierrez-Corona F, Loza-
Tavera H, Carlos-Tarres-Guzman M and Moreno-Sanchez R. 2001. Interaction of chromium with Microgenesis and plants. FEMS
Microbiology Reviews 25:335-347.
Chaudri AM, McGrath SP, Giller KE, Reitz E and Suerbeck DR. 1993. Enumeration of indigenous Rhizobium leguminosarum biovar
trifolii in soils previously treated with metal contaminated sewage
sludge. Soil Biology and Biochemistry .25:301–309
Chaoni A, Mazhoudi S, Groebal MH and Ferjani EI. 1997. Cadmium and zinc induction of lipid peroxidation and effects on antioxidant
enzyme activities in bean Phaseolus vulgaris L.. Plant Science 127:
139-147.
Chaudri AM, Allain CM, Barbosa-Jefferson VL, Nicholson FA, Chambers BJ and McGrath SP. 2000. A study of the impacts of Zn and Cu on
two rhizobial species in soils of a long term field experiment. Plant Soil 22:167–179.
Chen GH and Goldsbrough PB.1994. Increased activity of c- glutamylcysteine synthetase in tomato cells selected for cadmium
tolerance. Plant Physiology 106:233–239.
Chen YX. He YF, Yang Y, Yu YL, Zheng SJ, Tian GM, Luo YM and Weng MH. 2003. Effect of cadmium on soybean in contaminated
soils. Chemosphere.50:781-7.
Cheng SP. 2003. Effects of heavy metals on plants and resistance
mechanisms.Environmental Science and Pollution Research 10:256- 264.
Chugh LK and Sawhney SK. 1996.Effect of cadmium on germination, amylases and rate of respiration of germinating pea seeds.
Environmental Pollution 92:1-5.
Chugh, LK. and SK. Sawhney. 1999. Photosynthetic activities of Pisum sativum
seedlings grown in presence of cadmium. Plant Physiology
and Biochemistry.37: 297-303.
Clijsters H and Assche F. 1985. Inhibition of photosynthesis by heavy
metals. Photosynthesis Research 7:31–40.
Costa G and Morel JL. 1994. Water relations, gas exchange and amino acid content in Cd-treated lettuce. Plant Physiological and
Biochemistry 32:561-570.
Cox RM. 1988.The sensitivity of pollen from various coniferous and broad-leaved trees to combinations of acidity and trace metals.
New Phytologist 109: 193-201.
Cunningham SD, Shann JR, Crowley DE and Anderson TA. 1997. Phytoremediation of contaminated water and soil. Eds. Kruger
EL, Anderson TA and Coats JR. ACS symposium series 664, American Chemical Society, Washington.Pp2-19.
Dan T, Hale B, Johnson D, Conard B, Stiebel B and Veska E. 2008. Toxicity thresholds for oat Avena sativa L. grown in Ni-impacted
agricultural soils near Port Colborne, Ontario, Canada. Canadian Journal of Soil Science 88:389–398.
Das P, Samantaray S and Rout GR. 1997. Studies on cadmium toxicity
in plants:a review.Environmental Pollution 98:29-36.
De Carvalho MM, Edwards DG, Asher CJ and Andrew CS. 1982.Effects of aluminium on nodulation of two Stylosanthes species grown in
nutrient solution. Plant and Soil 64:141-152.
de Vries W, Lofts S, Tipping E, Meili M, Groenenberg JE and Schütze G. 2002. Impact of soil properties on critical concentrations of
cadmium, lead, copper, zinc, and mercury in soil and soil solution in view of ecotoxicological effects. Reviews of Environmental
Contamination and Toxicology 191:47–89.
Demirevska–Kepova K, Simova–Stoilova L, Stoyanova ZP and Feller U. 2006. Cadmium stress in barley: growth, leaf pigment, and
protein composition and detoxification of reactive oxygen species. Journal of Plant Nutrition 29:451-468.
Dewan MM and HR Dhingra. 2004. Cadmium partitioning and seed quality in two varities of pea and their hybrid as influenced by
rhizopheric cadmium. Indian Journal of Plant Physiology 9:15-20.
di Toppi LS and Gabbrielli R. 1999. Response to Cadmium in Higher
Plants.Environmental and Experimental Botany 41: 105-130.
Dixit V, Pandey V and Shyam R .2001.Differential oxidative responses to cadmium in roots and leaves of pea Pisum sativum
L cv.
Azad.Journal of Experimental Botany. 52: 1101-1109.
DraÛzkiewicz M, Sko _ ´rzyn´ska-Polit E , Krupa Z.2004. Copper- induced oxidative stress and antioxidant defence in Arabidopsis
thaliana
. BioMetals .17: 379–387.
Drolet G, Dumbroff EB, Legge RL and Thompson JE.1986.Radical
scavenging properties of polyamines.Phytochemistry.25:367-371.
Duan C and Wang H. 1995. Studies on the cell gene-toxicity of heavy metals to beans and micro-nuclear techniques. Acta Botanica Sinica
37
:14-24 In Chinese with English abstract. Ebbs SD and Kochian LV. 1997. Toxicity of zinc and copper to Brassica
species: implications for phytoremediation. Journal of Environmental Quality 26:776–781.
Egharevba and Omoregie H.2010. Effect of Cadmium on Seed Viability
of Vigna unguiculata, Ethnobotanical Leaflets:14:413-419.
Evans PT and Malmberg RL. 1989. Do polyamines have roles in plant development? Annual Review of Plant Physiology and Plant
Molecular Biology. 40:235–269.
Feng J, Shi Q, Wang X, Wei M, Yang F and Xu H. 2010. Silicon Supplementation Ameliorated the Inhibition of Photosynthesis and
Nitrate Metabolism by Cadmium Cd Toxicity in Cucumis sativus L. Scientia Horticulturae 123:521-530.
Finnegan PM and Chen W. 2010. Arsenic toxicity: The effects on
plant metabolism.Frontiers in Physiology.3:182-238
Flink I and Pettijohn DE. 1975.Polyamines stabilise DNA
folds. Nature. 253:62–63.
Kaur Nayyar : Heavy metal toxicity to food legumes: effects, antioxidative defense and tolerance mechanisms 1 5
Fontes RLS and Cox FR. 1998. Zinc toxicity in soybean grown at high iron concentration in nutrient solution. Journal of Plant Nutrition
21
:1723–1730. Foyer CH and Halliwell B. 1976. The presence of glutathione and
glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21-25.
Ghosh M and Singh SP. 2005.A review on phytoremediation of heavy metals and utilization of its byproducts. Applied Ecology and
Environmental Research 3:1-18.
Gimeno-García E, Andreu V and Boluda R. 1996.Heavy metals incidence in the application of inorganic fertilizers and pesticides to rice
farming soils.Environmental Pollution 92:19 –25.
Gill SS and Tuteja N. 2010. Reactive oxygen species and antioxidant machinery in abiotic stresstolerance in crop plants t Physiology
and Biochemistry.48 :909-930.
Gincchio R, Rodriguez PH, Badilla-Ohlbaum R, Allen HE and Lagos GE. 2002. Effect of soil copper content and pH on copper uptake of
selected vegetables grown under controlled conditions. Environmental Toxicology Chemistry 21:1736–1744.
Golan-Goldhirsh A,Mozafar A and Oerlli JJ. 1995. Effect of ascorbic acid on soybean seedlings grown on medium containing a high
concentration of copper. Journal of Plant Nutrition 18:1735-1741.
Gomez-Arroyo S, Cortés-Eslava J, Bedolla-Cansino RM, Villalobos- Pietrini R, Calderón-Segura ME and Ramírez-Delgado Y. 2001.Sister
chromatid exchanges induced by heavy metals in Vicia faba. Journal of Plant Biology 44:591-594.
Gonzalez-Chavez MC, Carrillo-Gonzalez R, Wright SF and Nichols KA. 2004. The role of glomalin, a protein produced by arbuscular
mycorrhizal fungi, in sequestering potentially toxic elements. Environmental Pollution 130:317–323.
Gupta YP. 1987. Anti-nutritional and toxic factors in food legumes:a
review. Plant foods for human nutrition.37:201-228.
Hall JL. 2001. Cellular mechanisms for heavy metal detoxification and
tolerance.Journal of Experimental Botany.56:1-11.
Han FX, Sridhar BBM, Monts DL and Su Y. 2004.Phytoavailability and toxicity of trivalent and hexavalent chromium to Brassica juncea
L
Czern. New Phytologist 162:489–499.
Harley H. and SE Smith.1983.Mycorrhizal symbiosis. Academic Press, London
Heckman JR, Angle JS and Chaney RL. 1987. Residual effects of sewage sludge on soybean II. Accumulation of soil and symbiotically fixed
nitrogen.Journal of Environmental Quality 16:117–124.
Hirsch PR, Jones MJ, McGrath SP and Giller KE. 1993. Heavy metals from past applications of sewage sludge decrease the genetic diversity
of Rhizobium leguminosarum biovar trifolii populations. Soil Biology and Biochemistry 25:1485–1490.
Hossain MA, Hasanuzzaman M and Fujita M. 2010. Up-Regulation of Antioxidant and Glyoxalase Systems by Exogenous Glycinebetaine
and Proline in Mung Bean Confer Tolerance to Cadmium Stress. Physiology and Molecular Biology of Plants16:259-272.
Huang CY, Bajaj FA and Vanderhoef LN. 1974. The inhibition of soybean
metabolism by cadmium and lead. Plant Physiology 54:122-124.
Israr M, Sahi S, Datta R and Sarkar D. 2006.Bioaccumulation and physiological effects of mercury in Sesbania drummonii.
Chemosphere 65:591–598.
Jamal A, Ayub N and Usman Mand Khan AG. 2002. Arbuscular mycorrhizal fungi enhance zinc and nickel uptake from
contaminated soil by bean and lentil. International Journal of Phytoremediation.4:205–221.
Johnson MS and Eaton J. 1980.Environmental contamination through residual trace metal dispersal from a derelict lead-zinc mine. Journal
of Environmental Quality 9:175–179.
Joschim HJ, Makoi R and Ndakidemi PA. 2009. The agronomic potential of vesicular-arbuscular mycorrhiza AM in cereals– legume
mixtures in Africa. African Journal of Microbiology Research.3 : 664-675.
Kamel HA. 2008. Lead accumulation and its effect on photosynthesis and free amino acids in Vicia faba grown hydroponically.Australian
Journal of Basic and Applied sciences.3:438-446
Kakkar, R.K. and Sawhney V.K. 2002.Polyamine research in plants- a
changing perspective. Phyiologia Plantarum 116 : 281-292.
Karina BB, Benavides MP, Gallego SM and Tomaro ML. 2003. Effect of cadmium stress on nitrogen metabolism in nodules and roots of
soybean plants. Functional Plant Biology 30:57–64.
Keilig K and Ludwig-Muller J. 2009. Effect of flavonoids on heavy metal tolerance in Arabidopsis thaliana
seedlings. Botanical Studies
50
:311-318. Kerkeb L and Kra¨mer U .2003.The role of free histidine in xylem
loading of nickel in Alyssum lesbiacum and Brassica juncea.Plant Physiology.131:716–724.
Khan AG, Kuek C, Chaudhry TM and Khoo CS, and Hayes WJ.2000.Role of plants, mycorrhizae and phytochelators in heavy metal
contaminated land remediation. Chemosphere 41:197–207.
Khan RM and Khan MM. 2010. Effect of varying concentration of nickel and cobalt on the plant growth and yield of chickpea. Australian
Journal of Basic and Applied Sciences 4:1036-1046.
Kristen U, Hoppe U and Pape W. 1993. The pollen tube test: a new alternative to draize eye irritation assay. Journal of the Society of
Cosmetic Chemists44:153-162.
Krupa Z and Baszynski T. 1995. Someaspects of heavy metals toxicity towards photosynthetic apparatus direct and indirect effects on
light and dark reactions. Acta Physiologie Plantarum 17:177–190.
Kukier U, Peters CA, Chaney RL, Angle JS and Roseberg RJ. 2004. The effect of pH on metal accumulation in two Alyssum species. Journal
of Environmental Quality 33:2090–2102.
Kumar G and Tripathi R. 2007. Lead induced cytotoxicity and
mutagenecity in Grass Pea. Turkish Journal of Biology 32:73-78.
Leyval C, Turnau K, Haselwandter K .1997. Effect of heavy metal pollution on mycorrhizal colonization and function: Physiological,
ecological and applied aspects. Mycorrhiza.7: 139–153.
Li HF, Gray C, Mico C, Zhao FJ and McGrath SP. 2009.Phytotoxicity and bioavailability of cobalt to plants in a range of soils.
Chemosphere 75:979–986.
Li Z, McLaren RG and Metherell AK. 2004. The availability of native and applied soil cobalt to ryegrass in relation to soil cobalt and
manganese status and other soil properties. New Zealand Journal of Agricultural Research 47:33–43.
Lin AJ, Zhang XH, Wong MH. 2007. Increase of multi-metal tolerance of three leguminous plants by arbuscular mycorrhizal fungi
colonization.Environmental Geochemistry and Health 29:473– 481.
Liu D, Jiang W and Gao X. 2003. Effect of cadmium on root growth, cell division and nucleoli in root tip cells of garlic. Plant Biology
47
:79-83.
1 6 Journal of Food Legumes 263 4, 2013
Liu D, Jiang W and Li M. 1992. Effects of cadmium on root growth and cell division of the root tip of garlic Allium sativum L.. Acta
Scientiae Circumstantiae 12:439-446 In Chinese with English abstract.
Madhava Rao KV, Sresty TVS .2000.Antioxidative parameters in the seedlings of pigeonpea Cajanus cajan
L. Millspaugh in response
to Zn and Ni stresses. Plant Sci. 157:113-128
Maksymiec 2007.Signaling responses in plants to heavy metal stress.Acta
Physiologie Plant arum.29:177–187.
Maksymiec W and Baszyn´ski T. 1996. Chlorophyll ûuorescence in primary leaves of excess Cu-treated runner bean plants depends on
their growth stages and the duration of Cu action. Journal of Plant Physiology 149:196–200.
Maksymiec W. 1997.Effect of copper on cellular processes in higher
plants. Photosynthetica 34:321–342.
Malik JA, Goel S, Sandhir R and Nayyar H. 2011. Uptake and distribution of arsenic in chick pea:Effects on seed yield and seed composition.
Communications in soil science and plant analysis. 42:1728-1738
Mandal SM and Bhattacharyya RN. 2007. Heavy metal toxicity on seed germination of four pulses. International Journal of Plant
Sciences 2:124-127.
Mascher R,B Lippmann,S Holzinger and H Bergmann. 2002. Arsenate toxicity:Effects on oxidative stress response molecules and
enzymes in red clover plants.Plant Science.163:961-969.
Mathys W. 1977.The role of malate, oxalate, and mustard oil glucosides in the evolution of zinc-resistance in herbage plants. Physiologia
Plantarum. 40:130–136.
McCarthy MC, Romero-Puertas JM, Palma LM, Sandalio FJ, CorpasM, Gómez LA and Del Río
.
2001. Cadmium induces senescence symptoms in leaf peroxisomes of pea plants. Plant, Cell and
Environment 24:1065–1073. Mcilveen WD and Cole HJr. 1974. Influence of heavy metals on
nodulation of red clover. Phytopathology 64:583-589.
Metwally A, Safronova VI, Belimov AA and Dietz KJ. 2005. Genotypic variation of the response to cadmium toxicity in Pisum sativum L.
Journal of Experimental Botany 56:167-178.
Mo W and Li M. 1992. Effects of Cd
2+
on the cell division of root tip
in bean seedlings. Bulletin of Botany 9:30-34 In Chinese with English abstract.
Moftah AE. 2000. Physiological response of lead polluted tomato and egg plant to the antioxidant ethylene diurea. Menufiya Journal of
Agricultural Research 25:933–955.
Molina AS, Nievas C, Chaca MVP, Garibotto F, Gonza´lez U, Marsa SM, Luna C, Gime´nez MS and Zirulnik F. 2008. Cadmium-induced
oxidative damage and antioxidative defense mechanisms in Vigna mungo
L. Plant Growth Regulation 56:285-295.
Moustakas MT, Lanaras L, Symeonidis S and Karataglis 1994. Growth and some photosynthetic characteristics of field grown Avena
sativa
under copper and lead stress. Photosynthetica 30: 389-396.
Mullineaux MP, Karpinski S and Bakerlogy NR. 2006. Spatial Dependence for Hydrogen Peroxide-Directed Signaling in Light-Stressed Plants.
American Society of Plant Biologists 141:346–350.
Mumthas S,Chidambaram AA, Sundaramoorthy P and Ganesh KS. 2010.Effect of Arsenic and Manganese on Root Growth and Cell
Division in Root Tip Cells of Green Gram Vigna radiata L.. Journal
of Agricultural and Food 22:
285– 297. Muneer S, Qadri TN, Mahmooduzaffarand
Siddiqi TO. 2011. Cytogenetic and biochemical investigations to study the response
of Vigna radiata to cadmium stress. African Journal of Plant Science 5:
183–192. Nieboer E and Richardson DHS.1980. The replacement of the non-
descript term “heavy metals” by a biologically and chemically significant classification of metals ions. Environmental Pollution
1:
3-26. Oladele EO, Odeigah PGC and Taiwo IA. 2013. The genotoxic effect
of lead and zinc on bambara groundnut Vigna subterranean. African Journal of Environmental Science and Technology 7:9-13.
Ouzounidou G, Giamparova M, Moustakas M and Karataglis S. 1995. Responses of maize Zea mays L. plants to copper stress. I. Growth.
Environmental and Experimental Botany 35:167–176.
Paivoke A. 1983. The long term effects of zinc on the growth and development, chlorophyll content and nitrogen fixation of the
garden pea Pisum sativum cv. Dipple Maj. Annales Botanici Fennici 20
:205-213. Paivoke AE and LK Simola. 2002. Arsenate toxicity to Pisum sativum:
Mineral nutrients,
chlorophyll content
and phytase
activity.Ecotoxicology and Environmental safety.49:111-121.
Pal SC. 1996. Effect of heavy metals on Legume-Rhizobium
symbiosis.Developments in Plant Soil Sciences.70:21-29
Panda SK, Singha NB and Khan MH.2003. Does aluminium phytotoxicity induce oxidative stress in green gramVigna radiata
Bulgarian Journal of Plant Physiology. 29:77–86
Pandey N and Sharma CP. 2002.Effect of heavy metals Co2+, Ni
2+
, and Cd
2+
on growth and metabolism of cabbage. Plant Science 163:753–
758. Parr PD and Taylor JrFG. 1982.Germination and growth effects of
hexavalent chromium in Orocol TL a corrosion inhibitor on Phaseolus vulgaris. Environment International 7:197-202.
Patra M and Sharma A. 2000.Mercury toxicity in plants. Botanical
Review 66: 379–422.
Pätsikkä E, Kairavuo M, Šeršen F, Aro EM and Tyystjärvi E. 2002. Excess copper predisposes photosystem II to photoinhibition in
vivo by outcompeting iron and causing decrease in leaf chlorophyll. Plant Physiology129: 1359-1367.
Peralta JR, Gardea-Torresdey JL, Tiemann KJ, Gomez E, Arteaga S and Rascon E. 2001. Uptake and effects of five heavy metals on seed
germination and plant growth in alfalfa Medicago sativa L. Bulletin of Environmental Contamination and Toxicology66:727-734.
Peterson CA and Rauser WE. 1979. Callose deposition and photoassimilate export in Phaseolus vulgaris exposed to excess
cobalt, nickel and zinc. Plant Physiology 63:1170-1174.
Piechalak A, Tomaszewska B, Baralkiewicz D, Malecka A.2002. Accumulation and detoxification of lead ions in legumes.
Phytochemistry
60: 153–162.