Effect of Varying Concentration of Nickel and Cobalt on the Plant Growth and Yield of Chickpea

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1 Austrlin Journl of Bsi nd Applied Sienes, 4(6): , 2010 ISSN Effet of Vrying Conentrtion of Nikel nd Coblt on the Plnt Growth nd Yield of Chikpe Mujeebur Rhmn Khn nd Mohd. Mhmud Khn Deprtment of Plnt Protetion, Fulty of Agriulturl Sienes, Aligrh Muslim University, Aligrh , Indi Abstrt: A pot experiment ws onduted to evlute the sensitivity of hikpe, Cier rietinum v. T-3 to nikel nd oblt t 0, 10, 50, 100, 200 nd 400 ppm onentrtions. The bterized seeds of hikpe were sown in 15 m ly pots ontining 1.5 kg stem sterilized soil. Before the sowing 550 ml metl solution of different onentrtions ws dded per pot. Nikel nd oblt t the higher onentrtions ( ppm) were found injurious to hikpe. Seed germintion, plnt growth nd biomss prodution were dversely ffeted. Root nodultion ws suppressed nd number of funtionl nodules ppreibly deresed. Lef hlorophyll ontents were gretly redued. These dverse impts refleted in redued yield of the plnts. Nikel ws pprently more hrmful thn oblt to hikpe. The study suggests tht ultivtion of rops like hikpe in metl polluted soils should be voided or pproprite ontrol mesures be dopted to mintin the hevy metl ontent of the soil below the dmge threshold levels, whih ws found to be 10 ppm Ni nd 50 ppm Co for hikpe. Key words: Biomss, hlorophyll ontent, hevy metls, plnt growth, root nodultion, seed germintion, soil pollutnt. INTRODUCTION Contmintion of soil in ultivted fields by industril effluents loded with toxi hevy metls hs emerged s new thret to griulture. Most of the effluents nd wstes ontin hevy metls in n mount suffiient enough to use toxiity to rop plnts (Huthinson nd Whiteby, 1974, Temple nd Bisessr, 1981; Khn et l., 2006). Exessive umultion of hevy metls like nikel, opper, oblt, et. in soil originting from metl mining nd proessing nd other tehnologil tivities of mn hve been reported on number of osions (Foy et l.,1978; Mliszewsk et l., 1958). Nikel is nturlly present in soil nd wter, usully in tre mounts. Severl plnts nutritionlly require -1 Ni for vrious metboli tivities (Welh, 1981). Ni (< 5 ìg L ) my stimultion of growth in higher plnts (Mishr nd Kr, 1974; Stedmn, 1968; Welh, 1981). However, rpid industriliztion nd urbniztion during the reent pst hve used umultion of Ni in vried hbitts where from the quisition by the plnts nd their further trnsfer to humn nd niml popultion my ffet the life forms seriously. Higher onentrtion of Ni my use numerous dverse effets on flor (Smith, 1996). Physiologil role of Ni nd its toxi effets on higher plnts hve been observed (Bisesser et l., 1983; Hle et l., 1985; Seregin nd Kozhevnikov, 2006; Khn et l., 2006). Growth of most plnts speies is dversely ffeted by tissue onentrtion bove 50 μg -1 Ni g dry weight. These effets re mnifested t morphologil, physiologil nd biohemil levels nd they my result either beuse of tendeny of Ni to ompete with other tions suh s C, Fe nd Zn nd thus to use their rtifiil defiienies. Higher levels of Ni in the soil nd in the plnts tissue often indue Zn or Fe defiieny whih leds to hrteristi symptoms of hlorosis (Anderson et l., 1973). Coblt is n essentil element for humns, nimls nd prokryotes, physiologil funtion for this element in higher plnts, however, hs not been identified. Coblt ontining vitmin B 12 does not our in -1 plnts. Norml Co onentrtions in plnts re ited to be s low s μg g dry weights nd its benefiil role s tre element neessry for the norml metboli funtions of the plnt hs been desribed (Bkkus et l., 2005). Higher onentrtion of this metl, however, my prove toxi nd severely interfere with physiologil nd biohemil funtions (El-Sheekh et l., 2003; Prmr nd Chnd, 2005 nd Jykumr -1 et l., 2008). Adverse effets of higher onentrtion of oblt (upto 750 ìg L ) hs been reported on some plnts (Terry 1981; Wlle, A. nd A.M. Abou-Zm Zm, 1989nd Jykumr nd Vijyrengn, 2006). Corresponding Author: Mujeebur Rhmn Khn, Deprtment of Plnt Protetion, Fulty of Agriulturl Sienes, Aligrh Muslim University, Aligrh , Indi, E-mil: mrkhn777in@yhoo.o.in 1036

2 Jykumr et l. (2008) observed tht seed germintion nd length of rdil nd plumule of rgi nd pddy -1-1 inresed signifintly t 5 ìg L, but gretly redued t ìg Co L. High onentrtions of Ni, Co nd other hevy metls disturb biologil equilibrium in the soil whih in turn unfvourbly influenes soil fertility, plnt development nd yield (Vllee nd Ulmer, 1972, Kbt- Pendis nd Pendis, 1973). Hevy metls lso interfere in biohemil retions of plnt nd indue physiologil disorders like redution in lef hlorophyll (Hle et l., 1985 nd Khn et l., 1988). Adverse effets of nikel, opper nd oblt hve been reported on growth hrters, biomss, yield, et. of elery, tomto nd lentil plnts both in mbient nd rtifiil tretment onditions (Bisessr et l., 1983, Hle et l., 1985, Khn et l., 2006). In the present pper, we report the growth performne of hikpe (Cier rietinum -1 L.) grown in soil rtifiilly treted with 0, 10, 50, 100, 200 nd 400 ìg L nikel nd oblt. MATERIALS AND METHODS Tretments nd Plnt Culture: Seed Germintion: Five onentrtions of nikel hloride (NiCl 2.5H2O) nd obltous hloride (CoCl 2.H 2O) viz. 10, 50, 100, 200 nd 400 ppm were prepred in distilled wter. Seeds of hikpe, Cier rietinum L. v. T-3 bterized with grm strin of Rhizobium were sown in offee ups of 6 m. dimeter ontining 100 g wter wshed nd sterilized snd (10seeds/up). In eh up 25 ml solution, the quntity found suffiiently enough for homogenous distribution of the metl solution ws dded prior to sowing. A set of ups reeived 25 ml distilled wter in ple of metl solutions served s ontrol. Eh tretment ws replited five times. The ups were pled on greenhouse benhes. Ten dys fter sowing the pots were exmined for seed germintion. Plnt Growth: To study the effet of nikel nd oblt on plnt growth nd produtivity of hikpe, 15 m ly pots were filled with 1.5 kg utolved soil (ly, snd, ompost 2:1:1). In eh pot 550 ml metl solution of different onentrtions ws dded seprtely prior to sowing. The mount (550 ml) ws found suffiient for the homogenous distribution of metl solution in 1.5 kg of soil. Pots reeiving distilled wter in ple of metl solution served s ontrol. Five seeds of hikpe v. T-3 bterized with ommeril Rhizobium of grm strin were sown in ll pots. Eh tretment ws replited five times. The pots were pled on greenhouse benhes. A week lter plnts were thinned to one in eh pot. At termintion, three months fter sowing, plnt growth (root nd shoot length), biomss (fresh nd dry weights of shoot nd root), root nodultion (funtionl, non-funtionl nd totl number of nodules/root system) nd yield (number of pods/plnt) were determined. Estimtion of Metls nd Lef Pigments in Plnt Tissue: Soil Anlysis: The soil smples were oven dried (40 C) nd finely ground (<0.1 mm). They were burnt to shes in ruible. Ashes of 1 g of soil smple were tken in onil flsk nd were moistened with 1 ml of doubledistilled wter; onentrted HCl nd HNO3 were suessively dded in 3: 1 rtio. The flsk ws heted gently on heting plte until the smple ws digested nd indited by the formtion of ler solution bove the soil residue. The mixture ws redued to volume of 1 ml. Double-distilled wter ws dded to reh finl filtrte volume of 100 ml nd filtered through Whtmn filter pper nos. 1 nd 42. Digested soil smples were nlyzed for Ni nd Co onentrtion by Atomi Absorption Spetrophotometer (Mlik nd Jiswl 2000). Nikel nd Coblt Assy: Plnt were dried in n hot ir oven t 80 C for 24 hr nd the leves, stem nd roots were grinded seprtely to mke orse powder. One grm of plnt dry mtter ws tken into 50 ml volumetri digestion tube. Using pipette 10 ml of the mixture HNO 3 nd HClO 4 (2:1, vv) ws dded nd swirled to thoroughly wet the smple. 25 mm reflux funnels were pled over smples nd llowed to predigest over night. The digestion tubes were pled into digestion blok port for 3 hours t 200ºC fter the HNO 3 fumes hve evolved. The funnels were removed 10 minutes before the end of the digestion. The tubes were tken out of the digestion blok, left for 20 minutes in hood. Therefter 10 ml deionized wter ws dded in the tubes on hot plte (90ºC). The ontents of the digestion tubes were mixed, ooled nd quntittively trnsferred into 25 ml volumetri flsk nd diluted to the finl volume, nd the Ni nd Co ontents of plnt digest were mde using Atomi Absorption Spetrophotometer (Miller, 1998). 1037

3 Chlorophylls: Chlorophyll, hlorophyll b nd totl hlorophyll were extrted by merting the lef tissue in 85 % etone (Lihenthler, 1987). One grm fresh lef tissue pooled from 5 plnts (45 dys old) of tretment ws put in mortr nd pestle, hlf filled with liquid nitrogen on ie. A smll mount of grinding snd ws lso dded. After the liquid nitrogen hving ompletely boiled off, the lef tissue ws grinded by the pestle. One ml of 85% etone (ph 8) ws dded in the mortr nd ground to mix. The suspension from mortr ws poured into 1.5 ml tube nd entrifuged t rpm for 3 minutes. One ml superntnt ws diluted to 3 ml with the etone nd bsorbne ws red in spetrophotometer t 637, 647 nd 663 nm. If bsorbne ws greter thn 1.0, the solution ws further diluted with etone. Sttistil Anlysis: The experiment ws performed over two onseutive yers. The generl trend in the effet of tretments on the onsidered vribles ws more or less identil during both yers of study but the effet of yers ws frequently signifint t P Hene the dt obtined from the five replites mintined eh yer were nlyzed seprtely. Results desribe the experiments onduted during seond yer. The dt on plnt growth, lef pigments nd Ni/Co umultion were subjeted to single ftor nlysis of vrine (ANOVA) nd lest signifine differenes (LSD) were lulted for eh vrible t probbility level of P 0.05, 0.01 nd (Dospekhov, 1984). The dt on metl umultion in plnt prts were nlyzed by the two wy single ftor ANOVA nd LSDs were lulted t the three probbility levels. Regression nlysis for some importnt vribles ws performed to lulte regression eqution nd presented in grphil form. Stndrd devition (SD) ws lulted for eh tretment nd hs been indited on the regression lines. Results: Seed Germintion: All the onentrtions of Ni nd Co used exept 10 ppm inhibited seed germintion (Tble 1). Perent germintions of seeds in the ontrol nd 10 ppm were t pr. But from 50 ppm Ni (P 0.01 nd P 0.001) nd 200 ppm Co onwrds, progressive nd signifint deline in the seed germintion ws notied in omprison to respetive ontrols (P 0.01). The differenes between 10 nd 50 ppm, 50 nd 100 ppm, 100 nd 200 ppm nd 200 nd 400 ppm of nikel, nd 100 nd 200 ppm nd 200 nd 400 ppm of oblt were lso sttistilly signifint (P 0.05 or P 0.01). Visul Symptoms: As generl indition of toxiity of nikel (Ni) nd oblt (Co), leves of hikpe developed hlorosis nd exhibited tip blkening nd redued size. Pre-mture lef drop lso ourred. The intensity of the symptoms ws found to be onentrtion dependent. The symptoms were most pronouned in tretments with 400 ppm onentrtions of the metls. Roots of the metl treted plnts showed limited growth nd their extensiveness ws heked, lterl roots being more suppressed. Disolourtion of roots beuse of development of drk brown pthes ws notied in plnts treted with higher onentrtions of Ni nd Co. In overll ssessment, Ni ws more injurious thn Co. Plnt Growth, Biomss Prodution nd Pod Formtion: Both Ni nd Co prtiulrly t higher onentrtions used dverse effets on plnt growth. Plnts treted with the metl solution exhibited poor root nd shoot growth with less number of brnhes (Tble 2). A signifint redution ourred in shoot length t 200 nd 400 ppm Ni nd 400 ppm Co over ontrol (P 0.05). Differene in shoot lengths obtined with 200 nd 400 ppm Ni ws lso signifint (P 0.05). Redutions in shoot growth with other onentrtions of either metl were not signifint. Root growth of hikpe plnts treted with 100, 200 nd 400 ppm Ni nd 200 nd 400 ppm Co ws signifintly retrded in omprison to respetive ontrols (Tble 2). Growth of lterl roots ws gretly suppressed. Biomss (fresh nd dry weights of root nd shoot) of hikpe ws impeded by the metl tretments (Tble 2). Fresh weights of root nd shoot were more ffeted by Ni thn Co. There ws signifint redution in the fresh weight of root of the plnts grown in 100, 200 (P 0.05) nd 400 ppm Ni (P 0.01) or 400 ppm Co treted soils (P 0.05). Signifint redution in fresh weight of shoot ourred with 400 ppm of Ni nd Co (P 0.05). Similr redution in dry weight of root ws notied with the pplition of 200 nd 400 ppm of Ni (P 0.01) nd 400 ppm of Co (P 0.05) in omprison to untreted plnts (ontrol). Higher onentrtions (100, 200 nd 400 ppm) of both the metls were signifintly effetive in reduing the dry weights of hikpe shoots (P 0.05). All the three onentrtions were eqully hrmful s redutions used by the onentrtions did not differ signifintly t P

4 Tble 1: Effet of nikel nd oblt on perent seed germintion of hikpe (men of two sesons) Tretments Seed germintion % ìg Ni nd Co /L Nikel Coblt b b L.S.D P P P b b Eh vlue is men of five replites signifintly different from the ontrol t P 0.01 ( ) nd P Tble 2: Effet of soil pplition of nikel nd oblt on the plnt growth of hikpe (men of two sesons). Tretment Nikel effets Coblt effets Ni/Co ìg/l Length (m) Fresh weight (g) Dry weight (g) Yield Length (m) Fresh weight (g) Dry weight (g) Yield Root Shoot Root Shoot Root Shoot Root Shoot Root Shoot Root Shoot 0 (ontrol) b b b b L.S.D. P P P b Eh vlue is men of five replites. b signifintly different over ontrol t P 0.05 ( ) nd P 0.01 ( ) nd P 0.001( ) Pod formtion ws gretly suppressed by both the metls, Ni being more hrmful thn Co nd overll it ws onentrtion dependent. Tretments with 100, 200 (P 0.05) nd 400 ppm Ni (P 0.01) nd 200 nd 400 ppm Co (P 0.05) used signifint deline in the pods/plnt over untreted plnts. Root Nodultion: Root nodultion ws suppressed by Ni nd Co onentrtions showing orresponding derese in totl number of nodules with n inrese in the onentrtions of either metl. This impt ws, however, signifint with 200 nd 400 ppm of Ni nd 400 ppm of Co over ontrol (Tble 3). Number of non-funtionl nodules ws not ffeted but number of funtionl nodules delined onsiderbly. The dverse impt on funtionl nodules ws signifint with 200 (P 0.05) nd 400 ppm Ni (P 0.01) nd 400 ppm Co (P 0.05). Tble 3: Effet of soil pplition of nikel nd oblt on root nodultion of hikpe (men of two sesons) Tretments Nikel Coblt Funtionl Non funtionl Totl Funtionl Non funtionl Totl nodule nodule nodule nodule nodule nodule b LSD P P P b b Eh vlue is men of five replites. signifintly different over ontrol t P 0.05 ( ) nd P 0.01 ( ) nd P 0.001() Lef Pigments: A grdul nd signifint redution in the totl hlorophyll ontent ourred in the plnts treted with 50, 100 (P 0.05), 200 (P 0.01) nd 400 ppm Ni (P 0.001) over ontrol (Tble 4). But this redution with Co ws signifint t only 200 (P 0.05) nd 400 ppm (P 0.01). The signifint redution in hlorophyll ourred in the plnts treted with 100, 200 (P 0.05) nd 400 ppm Ni (P 0.01) nd 200 nd 400 ppm Co (P 0.05). Ni ws omprtively more inhibitory thn Co for hlorophyll. Signifint redution in the hlorophyll b ws reorded with only 200 nd 400 ppm Ni nd not with ny onentrtion of Co (P 0.05). 1039

5 Tble. 4: Effet of soil pplition of nikel nd oblt on lef hlorophyll of hikpe (men of two sesons) Tretments Nikel Coblt Chl Chl b Totl Chl Chl Chl b Totl Chl b b b L.S.D P P P b Eh vlue is men of five replites. signifintly different over ontrol t P 0.05 ( ) nd b P 0.01 ( ) nd P 0.001( ) Fig 1: Correltion ship between metl umultion in plnt tissue (root nd shoot) nd dry weight of shoot nd root of hikpe Metl Aumultion: Soil: Applition of 550 ml metl solution of different onentrtions resulted to orresponding inrese in the onentrtion of the respetive metls in the pot soil (Fig 3). Conentrtion of both Ni nd Co dy fter pplition (pre-plnting) ws found signifintly greter thn the pplied onentrtion of the solution (P 0.05). The soil onentrtion of the two metls t hrvest (4 month fter pplition), however, slightly deresed in omprison to the pre-plnting onentrtion (Fig 3). 1040

6 Fig. 2: Effet of soil pplition of nikel nd oblt on hlorophyll, hlorophyll b nd totl hlorophyll of hikpe leves Plnt Prts: Highest onentrtion of the hevy metls ws reorded in the leves of hikpe (Fig 4). Ni onentrtion in the leves of plnts pplied with 10 ppm onentrtion ws 7.8 ìg/g of lef tissue whih inresed grdully with the inrese in the onentrtion of metl solution pplied. A strong liner reltionship ws reorded with the onentrtion of metl solution pplied nd its umultion in the leves (Fig 4). The onentrtion of Ni in the roots ws signifintly less thn leves t 10, 50 nd 100 ppm tretments (P 0.05). Lowest onentrtion of Ni ws reorded in stem nd ws signifintly less thn root (P 0.001). But stem onentrtion of Ni lso showed strong liner reltionship between onentrtion of metl solution pplied nd its umultion in stem. Coblt onentrtion in plnt prts lso inresed orrespondingly with the inresed in metl onentrtion pplied (Fig 4). Greter umultion of Co ws reorded in leves followed by roots nd lowest in stem. The liner reltionship between onentrtion of Co in the solution pplied nd umultion of the metl in plnt prts ws invribly reorded in leves, stem nd roots. 1041

7 Fig. 3: Correltion ship between soil pplition of hevy metls nd their umultion in soil Disussion: Contmintion of soil with hevy metls hs found to suppresses to the seed germintion (Jin, 1987; Khn et l., 1987, 1988), s observed in the present study. Prominent visible injury viz. shoot injury, lef hlorosis, lef-tip blkening nd root injury with restrited lterl growth nd slight disolourtion, the symptoms observed on hikpe due to Ni nd Co toxiity re omprble to those ppered on plnts like Aer rubrum, Cornus stolonifer, Lonier ttri, Pinus resinos nd Lne ulnens due to Ni, Co nd Cu toxiity (Hele nd Ormrod, 1982; Khn et l., 1987, 1988, Hilmy nd Gd, 2002, Ynqun et l., 2004, Ynqun et l., 2005). These symptoms espeilly the yellowing, hlorosis nd nerosis my hve ourred due to injury or denturing of hlorophyll moleules (Smith, 1996) or rtifiil defiieny of Fe, Zn et. indued by Ni nd Co (Anderson et l., 1973, Esrre et l., 2000, Hilmy nd Gd, 2002, Hilmy et l., 2002, Ynqun et l., 2004, Ynqun et l., 2005). Diminutive effets of hevy metls on plnt growth prtiulrly on shoot hve been reorded ner nikel refinery where Ni nd Co were present t higher onentrtions (Bisessr et l., 1983, Gbriell nd Attil, 2002, Rotkittikhun et l., 2006). Redution in shoot growth of lentil plnts by Ni, Cu nd Co in the rtifiil tretments hve been observed by Khn et l. (1987, 1988). Although the roots were diretly exposed to the 1042

8 Fig. 4: Aumultion orreltion between metl tretments nd Nikel/Coblt umultion in root, stem nd leves of hikpe. metl but they t s trnsporter by bsorbing the metls nd trnsporting it to the shoot where the metl gets umulted s evidened by reltively greter onentrtion of Ni nd Co in lef thn root. The metl, however, used toxiity to both the prts. Suppression of root enlongtion nd seedlings of Pie bies, elery nd tomto plnts due to vrious hevy metls on extensiveness nd prolifertion of root nd their subsequent effets on shoot growth led to suppression of dry mtter prodution (Bisessr et l., 1983; Hke et l., 1985; Reddy, 2003, Yng et l., 2003, Vnik et l., 2005, Khn et l., 2006). Adverse effets of Ni/Co on the plnt growth were onentrtion dependent. Greter umultion of the metl in leves or roots resulted to orresponding derese in their growth or dry weights. Regression nlysis hs lso reveled negtive reltionship between metl umultion in the root nd leves nd subsequent derese in their dry weight nd yield. Over ll greter toxi effets of Ni thn Co s observed in the present study hve lso been reported by other workers (Froster, 1954, Hele nd Ormrod 1982, Bisessr et l., 1983, Hle et l., 1985, Jsiewiz nd Antonkiewiz, 2000, Pndey nd Shrm, 2002, Giridhr nd Siddrmpp, 2002, Hilmy nd Gd, 2002, Hilmy et l., 2002, Ogundirn, 2007). Root nodultion is diretly relted to nitrogen eonomy of leguminous plnts nd ffets their produtivity. Poor root nodultion on hikpe plnts grown in Ni or Co ontminted soil my hve interfered in the nitrogen-fixing system using diret toxi effet on Rhizobium nd/or inhibiting leghemoglobin synthesis (Khn et l., 1987, Bunzl et l., 2001, Boulrbh et l., 2006). Hevy metls my lso denture hlorophyll pigments or my ffet their synthesis (Hle et l., 1985, Hilmy nd Gd, 2002, Hilmy et l., 2002, Boulrbh et l., 2006). In the present study Ni nd Co used suppression of hlorophylls ontents of leves. Metl toxiity my hve used phephytiniztion of hlorophyll moleules whih ws expressed s browning nd 1043

9 + nerosis of lef tissue. Metl ions my reple Mg ion from the tetrpyrol ring of the hlorophyll moleule leding to denturing of the pigments s evident from the signifint redution in lef ontents of hlorophyll (Hilmy nd Gd, 2002, Hilmy et l., 2002, Reddy, 2003, Khn et l., 2006, Boulrbh et l., 2006, Ogundirn, 2007). Present study hs reveled 10 nd 50 ppm s dmge threshold level of Ni or Co respetively for hikpe ultivtion. Approprite efforts should be employed to mintin the metl onentrtion below the dmging level in the res lose to nikel or oblt soure. ACKNOWLEDGEMENTS Finnil ssistne from University Grnt Commission, Government of Indi, New Delhi through mjor reserh projet No /2006(SR) is grtefully knowledged. REFERENCES Agrwl, S.C., S.S. Bisht, C.P. Shrm nd A. Agrwl, Effet of defiieny of ertin mironutrients on the tivity of letolsp in rdish plnts grown in S nd P ulture. Cn. J. Bot., 54: Anderson, J.G., D.R. Myer nd R.K. Myer, Hevy metl toxiity levels of Nikel, oblt nd hromium in the soil nd plnts ssoited with visul symptom nd vrition in growth of n ot rop. Aust. J. Agri. Res., 24: Bkkus, E., B. Gouget, J.P. Gllien, H. Khodj, F. Crrot, J.L. Morel nd R. Collins, Conentrtion nd distribution of oblt in higher plnts: The use of miro-pixe spetrosopy Nul. Instr. Meth. Phys. Res. B. 231, 350. Bisessr, S., R.J. Rinne nd J.W. Potter, Effet of hevy metl nd Meloidogyne hpl on elery grown on orgni soil ner nikel refinery. Plnt Diseses, 67(1): Boulrbh, A., C. Shwrtz, G. Bitton, W. Aboudrr, A. Ouhmmou nd J.L. Morel, Hevy metl ontmintion from mining sites in South Moroo: 2. Assessment of metl umultion nd toxiity in plnts, Chemosphere, 63: Bunzl, B., M. Trutmnnsheimer, P. Shrmel nd W. Reifenhuser, Avilbility of Arseni, opper, Led, Thllium nd Zin to vrious vegetbles grown in slg-ontminted soils, J. Environ. Qul., 30: Crooke, W.M., Effet of soil retion on uptke of Nikel from serpentine soil. Soil Si., 81: Dospekhov, B.A., Field experimenttion. Mir Publishers. Mosow, Russi, pp: 352. El-Sheekh, M.M., A.H. El-Nggr, M.E.H. Osmn nd E. El-Mzly, Effet of oblt on growth, pigments nd the photosyntheti eletron trnsport in Monorphidium minutum nd Nitzhi perminut. Brz. J. Plnt Physiol., 15: 159. Esrre, J., C. Lefebvre, W. Gruber, M. Lebln, L. Leprt, Y. Riviere nd B. Dely, Zin nd dmium hyperumultion by Thlspi erulesens from metlliferous nd nonmetlliferous sites in the Mediterrnen re: implitions for phytoremedition, New Phytol., 145: Foy, C.D., R.L. Chney nd M.C. White, The physiology of metl toxiity in plnts. Annul Review plnt physiology, 29: Froster, W.A., Toxi effets of hevy metls on rop plnts grown in soil ulture. Annul Applied Biology, 41: Gbriell, M. nd A. Attil, Hevy metl uptke by two rdish vrieties, At Biol. Szegediensis, 46(3-4): Giridhr, M. nd R. Siddrmpp, Effet of hevy metls on urese tivity in soil. Curr. Res. Univ. Argri. Si. Bnglore, 31: 4-5. Godbold, D.L. nd A. Huttermnn, Effet of zin, dmium nd merury on root elongtion of Pie bies (Krst) seedlings nd the signifine of these metls to forest diebk. Environmentl Pollution, 38(4): Hele, E.L. nd D.P. Ormrod, Effet of nikel nd opper on Aer rubrum, Cornus stolenifer, Lonier ttri nd Pinus resinos. Cndin journl of Botny, 60: Hele, E.L., D.P. Ormrod, P.J. Lffey nd O.B. Allen, Effet of nikel nd opper mixtures on tomto in snd ulture. Environtl Pollution, (A) 39:

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