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Effects of Cadmium on Morphology, Photosynthesis and Protein Profile of Bean Plants Pournia, Farnaz 2009-03-14

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Ef f ect s of Cadmi um on Mor phol ogy, Phot osynt hesi s and Pr ot ei n Pr of i l e of Bean Pl ant s Farnaz Pournia; Jarnail Mehroke; Santokh Singh Department of Botany, Biology Program, University of British Columbia, Vancouver, B.C, Canada  Abst r act Cadmi um ( Cd) , a c anc er - c aus i ng heav y met al i on, c onc ent r at i on i n t he env i r onment has r ec ent l y s hown a r api d i nc r eas e mai nl y as a r es ul t of human ac t i v i t y . Si nc e many pl ant s ar e us ed as medi c i ne and f ood, i t i s c r uc i al t o i nv es t i gat e ef f ec t s of Cd on pl ant gr owt h and dev el opment . Thi s s t udy ex ami nes t he t i me c our s e and c onc ent r at i on- dependent ef f ec t s of Cd on mor phol ogy , phot os y nt hes i s and pr ot ei n pr of i l es in hy dr oponi c al l y - gr own bean pl ant s ( Phas eol us v ul gar i s ) . Hi gh c onc ent r at i ons of Cd ( 1000 and 100 uM) i nduc ed l eaf wi l t i ng and r educ ed bot h l eaf gr owt h and phot os y nt hes i s r at e wi t hi n 24 and 48 hour s r es pec t i v el y . Lower c onc ent r at i on of 10 uM Cd s howed t he wi l t ed mor phol ogy af t er 96 hour s . In addi t i on, t he Cd- i nduc ed c hanges i n al pol l ut pr ot ei Env n i rpronm of ent i l es es pec i ali on ly thas he iphot eas ed hes dr iam i c al l syt rov t y nt s at and eser s tprheot pas ei ns . nc r os fwiew y bec e at ofi on human l l dec beades di sm cai usnl s ed i n aus r el to ac i v i t ogy i es , ( 1)l . eaf Pr es enc e h of and Cd mort phol gr owt cphot aus es f f er t y pespl ant of spr os y ntdihes i sent i n bean . obl ems t o l i v i ng bei ngs i nc l udi ng a wel l k nown c anc er c aus at i on i n human ( 2) . Si nc e pl ant s as pr i mar y pr oduc er s ar e c ons umed by ani mal s and human and al s o bec aus e of t he us age of pl ant s i n dr ug pr oduc t i on, i t i s nec es s ar y t o s t udy t he ef f ec t s of Cd on pl ant s ( 1) . Pr ev i ous s t udi es hav e s hown negat i v e ef f ec t s on mor phol ogy and r educ ed phot os y nt hes i s and t r ans pi r at i on r at e i n pr es enc e of Cd ( 2) . Thi s has been at t r i but ed t o t he i nt er f er enc e of t he el ement wi t h mol ec ul ar pat hway s s uc h as phot os y s t em II and el ec t r on t r ans por t c hai n in t hy l ak oi d membr ane ( 2) . Al s o, s t omat al c l os ur e i n Cd s t r es s c ondi t i ons r es ul t s i n l ower t r ans pi r at i on r at e and l ower CO2 i nt ak e whi c h i n t ur n c aus es l ower phot os y nt hes i s r at e ( 2) . Cd i s al s o k nown t o di s t ur b t he s t r uc t ur e and ac t i v i t i es of enz y mes and al t er t he l i pi d c ompos i t i on and per meabi l i t y of membr ane ( 3) . Fi nal l y ac c umul at i on of Cd i n pl ant or gans eens hoot Beansand( Phas ul rgar i se ) s uc h Gras r ooteolmus ay iv nt oduc wer own he gr e s fes or of7 i nhie bi gr t or s ti on rtegul at eenhous e pr oc es day s . s sThe ei l att ihen pl ant uc h ass eedl ni t irngs at e aswer si m on tpat r ans f er (r 3) ed . ont o a hy dr oponi c s y s t em hway whi c h i s a gr owi ng medi um wi t h k nown c onc ent r at i ons of nut r i ent s . Af t er 7 m or e obj dayec s t (i 16 i ght day ) i, s t the The v e hr of. tl hi s s/t udy o pl h i Cd ex ant ami sne wer t hee eft rf eat ec t ed of wi ct adm um at on cmonc ent rogy at i, ons of 10, phot 100 or phol os yand nt hes1000 i s, uM . t r ans pi r at i on, and t he pr ot ei n  Fi gur es 6 hr  24 hr  48 hr  96 hr  Resul t s and Mor phol ogy Di scussi on  168 hr  • Hi gh c onc ent r at i ons of Cd, 1000 and 100 uM i nhi bi t gr owt h and dev el opment of bean pl ant s and c aus e wi l t ed mor phol ogy i n 24 hour s and 48 hour s r es pec t i v el y • The ef f ec t s of l ow Cd c onc ent r at i on ( 10 uM) ar e obs er v ed af t er 96 hr • Cd ef f ec t s ar e bot h c onc ent r at i on- dependent .  Fi gur e 1 - Ti me Cour s e Changes i n and Cd Tr eat ed bean pl ant s  • Phot os y nt hes i s is par t i al l y ( appr ox i mat el y 50%) r educ ed in hi gher Cd c onc ent r at i ons , 100 and 1000 uM, af t er 24 hr  Mor phol ogy of Cont r ol  1 .8 Cont rol  www.PosterPresentations.com  1 0 0 uM Cd  s)  1 0 0 0 uM Cd  T ra nspira t ion Ra t e (um ol.m  5  1 0 0 uM Cd  -2  1 0 uM Cd  6  4 3 2 1 0 0  24  48  96  1 0 uM Cd  1 .6  Cont rol  -2  -1  Phot osynt he sis Ra t e (um ol.ms )  7  1 .4  1 0 0 0 uM Cd  1 .2 1  • Tr ans pi r at i on is r educ ed ( appr ox i mat el y 75%) i n 100 and 1000 uM Cd by 24 hr and i n 10 uM Cd by 96 hr • Tr ans pi r at i on is c ompl et el y i nhi bi t ed by 1000 uM Cd af t er 96 hr  0 .6 0 .4  0 0  Fi gur e 2 - Ti me Cour s e Changes i n phot os y nt hes i s Rat e ( umol . m- 2 s - 1 ) of Cont r ol and Cd Tr eat ed bean pl ant s  • Cd c ompl et el y i nhi bi t ed phot os y nt hes i s r at e at and af t er 96 hr  0 .8  0 .2  168  T im e (hr.)  24  48  96  168  Tim e (hr.)  Fi gur e 3 - Ti me Cour s e Changes i n Tr ans pi r at i on Rat e ( umol . m- 2 s 1 ) of Cont r ol and Cd t r eat ed bean pl ant s  •A mor e r api d dec l i ne in phot os y nt hes i s r at e c ompar ed to t r ans pi r at i on, s ugges t s t hat f ac t or s ot her t han s t omat al c l os ur e ar e af f ec t i ng phot os y nt hes i s . Thes e c oul d be due t o c hanges i n s t r uc t ur e and/ or f unc t i on of phot os y nt het i c pr ot ei ns  Pr ot ei n Pr of i l e St andar d Pr ot ei n s 66 KDa  St andar d Pr ot ei n s66 KDa  Lar ge subuni t of Rubi sco and ATPase ( 54 KDa) LHC- I I B  45 KDa Lar ge subuni t of Rubi sco & ATPase ( 54KDa)  45 KDa 36 KDa 29 KDa  36 KDa 29 KDa  LHC- I I B ( 25 KDa)  24 KDa 20 KDa  Newl y synt hesi sed pr ot ei n ( 18 KDa)  14. 2 KDa 24 hr  24 hr  Cont r o l  10 uM  24 hr 100 uM  Mat er i al s and Met hods  POSTER TEMPLATE BY:  and  Phot osynt hesi s and Tr anspi r at i on Rat es  I nt r oduct i on  pr of i l es . Gr owt h and dev el opment of pl ant s wer e moni t or ed f or s ev en day s . Rat es of phot os y nt hes i s and t r ans pi r at i on f or eac h t r eat ment wer e meas ur ed at t he end of 4, 6, 24, 48, 96 and 168 hour s af t er t r eat ment us i ng LI - 6200 Por t abl e Phot os y nt het i c Sy s t em. By t he end of eac h s pec i f i ed per i od, l eav es f r om c ont r ol and t he t hr ee  t i me-  Cont r ol  24 hr  48 hr  48 hr  48 hr  48 hr  1000 uM  Cont r o l  10 uM  100 uM  1000 uM Cont r o l  10 uM  100 uM  1000 uM  Cont r ol  10uM 100 uM  1000 uM  6 hr  24 hr  uM  uM  ol  uM  uM  96 hr  96 hr  96 hr  100 uM  1000 uM  10 uM  uM  168 hr  168 hr  Cont r o l  10 uM  Fi gur e 5 - Pol y ac r y l ami de gel s howi ng t he maj or pr ot ei n bands i n 24 and 48 hr c ont r ol and Cd t r eat ed bean pl ant s Cont r ol  10 uM  100 uM 1000 uM  96 hr  Fi gur e 6 - Wes t er n Bl ot , Dehy dr i n ( 60 KDa) i n 4 and 6 hr c ont r ol and Cd t r10eat 100 ed bean plr ant s 100 1000 10 1000 Cont uM  Newl y synt hesi sed pr ot ei n ( 18 KDa)  96 hr  4 hr  Cont r ol  20 KDa  14. 2 KDa  Fi gur e 4 - Pol y ac r y l ami de gel s howi ng t he maj or pr ot ei n bands i n 24 and 48 hr c ont r ol and Cd t r eat ed bean pl ant s  0 hr  ( 25 KDa)  24 KDa  Cont r ol  10 uM  168 hr  Fi gur e 8 - Wes t er n Bl ot Dehy dr i n ( 60 KDa) i n 96 and 168 hr c ont r ol and Cd t r eat ed bean Cont r 10 uM 100 uM 1000 pl ant s ol uM  Concl usi ons  • Appear anc e of t he 18 KDa pr ot ei n, a l i k el y Cd s t r es s r es pons e pr ot ei n, 1. or phol c al and and c phy s i olr ogi al is M bot h t ogi i meonc ent at i conef f ec t s of Cd ar e bot h t i me- and dependent c onc ent r at i on- dependent . • Dehy dr i n pr ot ei n whi c h i s a s t r es s 2. y nt hes i secand r ans r at inson pr ot Phot ei n os and pr ot t s plt ant s piagai t rdehy at es ar ie on i nhii sbi tsed by Cd dr at t r ongl y pr es ent i n Cd ment s profot1000 at 48 hr and 3. t rAeat new ei n uM ( 18 KDa) is of 10 t ed uM iatn rand af t e er t o 96 Cd hr s t r es s . det ec es pons  Fut ur e St udi es • Rubi s c o and LHC- I I B pr ot ei n l ev el s  •ar e I dent r uc t ur eby and t i oner of noti f yaf sf tec t ed Cd; f unc howev ,  tt he hei r18 acKDa t i v iprt ot y ei mi n ght be •• CdSt udy t sl ak atoi dt he mm em ane af f Cd ec t sef f ec t hy em brbrane per ev elc h may l ead t o a per m meabi eabi ll ii tt yy l whi dec l i ne i n pr ot on gr adi ent ac r os s • I nv es t i gat e Cd ef f ec t s on gene texhe br ane and a r educ t i on i n pr esmsem i on Iphot woulos d lyint k e hes t o ti hank my eBi ol ogy 352 l ab par t ner s s r at  Acknowl edgement s  Br eanne J ohns on, Samant ha I v er s en and Ros anne Pi puni c f or hel p wi t h a pr el i mi nar y ex per i ment . I woul d al s o l i k e t o t hank Shi ng Zhan f or hel pi ng wi t h s ome dat a c ol l ec t i on. Fi nal l y , I woul d l i k e t o t hank Sk y l i ght Dev el opment Fund, Fac ul t y of Sc i enc e and t he Depar t ment of Bot any f or f i nanc i al s uppor t .  Ref er ences  48 hr  Fi gur e 7 - Wes t er n Bl ot , Dehy dr i n ( 60 KDa) i n 24 and 48 hr c ont r ol and Cd t r eat ed bean pl ant s  • A new pr ot ei n ( 18 KDa) appear s i n bean l eav es t r eat ed wi t h 1000 uM Cd at and af t er 48 hr , i n 100 uM at and af t er 96 hr and i n 10 uM at 168 hr  Fi gur e 9 - Wes t er n Bl ot LHC- I I B ( 25 KDa) i n 48 hr c ont r ol and Cd t r eat ed bean pl ant s  1.  Rai V. , Khat oon S. , Bi sht S. S. and Mehr ot r a S. 2005. Ef f ect s of Cadmi um on gr owt h, ul t r amor phol ogy of l eaf and secondar y met abol i sm of Phyl l ant hus amar us and Thor n. Chemospher e. 61 : 1644- 1650  2.  Cl i j st er s H. , Van Assche F. 1985 I nhi bi t i on of Phot osynt hesi s Resear ch. 7: 31- 40, 1985.  3.  Sandal i o, L. M. , Dal ur z o, H. C. , Gomez , M. , Romer o- Puer t as, 2001. Jour nal of Exper i ment al Bot any 52: 2115- 2126.  phot osynt hesi s by heavy met al s. M. C.  and del  Ri o,  L. A.  

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