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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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POSTER TEMPLATE BY: www.PosterPresentations.com 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 . Fi gur es Resul t s and Di scussi on Mat er i al s and Met hods I nt r oduct i on Concl usi ons Ref er ences    Cadmi um ( Cd) ,  a cancer - causi ng heavy met al  i on,  concent r at i on i n t he envi r onment  has r ecent l y  shown a r api d i ncr ease mai nl y as a r esul t  of human act i v i t y .  Si nce many pl ant s ar e used as medi c i ne and f ood,  i t  i s cr uci al  t o i nvest i gat e ef f ect s of  Cd on pl ant  gr owt h and devel opment . Thi s st udy exami nes t he t i me cour se and concent r at i on- dependent  ef f ect s of  Cd on mor phol ogy,  phot osynt hesi s and pr ot ei n pr of i l es i n hydr oponi cal l y- gr own bean pl ant s ( Phaseol us vul gar i s ) .  Hi gh concent r at i ons of  Cd ( 1000 and 100 uM)  i nduced l eaf  wi l t i ng and r educed bot h l eaf  gr owt h and phot osynt hesi s r at e wi t hi n 24 and 48 hour s r espect i vel y.  Lower  concent r at i on of 10 uM Cd showed t he wi l t ed mor phol ogy  af t er  96 hour s.  I n addi t i on,  t he Cd- i nduced changes i n pr ot ei n pr of i l es especi al l y  t he phot osynt hesi s and st r ess pr ot ei ns wi l l  be di scussed i n r el at i on t o mor phol ogy,  l eaf  gr owt h and phot osynt hesi s i n bean pl ant s.    Gr een Beans ( Phaseol us vul gar i s) wer e gr own  i n t he gr eenhouse f or  7 days.  The seedl i ngs wer e t hen t r ansf er r ed ont o a hydr oponi c syst em whi ch i s  a gr owi ng medi um wi t h known concent r at i ons of  nut r i ent s.  Af t er  7 mor e days ( 16 hr .  l i ght  /  day) ,  t he pl ant s wer e t r eat ed wi t h Cd at concent r at i ons of  10,  100 and 1000 uM. Gr owt h and devel opment  of  pl ant s wer e moni t or ed f or  seven days.  Rat es of  phot osynt hesi s and t r anspi r at i on f or  each t r eat ment  wer e measur 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 osynt het i c  Syst em.  By t he end of  each speci f i ed per i od, l eaves f r om cont r ol  and t he t hr ee t r eat ment s wer e col l ect ed f or pr ot ei n [ Ri bul ose- 1, 5- bi phosphat e car boxyl ase/  oxygenase ( Rubi sco) , Li ght  Har vest i ng Compl ex- I I B ( LHC- I I B)  and Dehydr i n) ]  anal ysi s  by SDS- PAGE and West er n bl ot  t echni ques. Mor phol ogy • Hi gh concent r at i ons of  Cd,  1000 and 100 uM i nhi bi t  gr owt h and devel opment  of  bean pl ant s and cause wi l t ed mor phol ogy i n 24 hour s and 48 hour s r espect i vel y • The ef f ect s of  l ow Cd concent r at i on ( 10 uM)  ar e obser ved af t er  96 hr • Cd ef f ect s ar e bot h t i me-  and concent r at i on- dependent . Phot osynt hesi s and Tr anspi r at i on Rat es • Phot osynt hesi s  i s  par t i al l y ( appr oxi mat el y  50%)  r educed i n hi gher  Cd concent r at i ons,  100 and 1000 uM,  af t er  24 hr •  Cd compl et el y i nhi bi t ed phot osynt hesi s r at e at  and af t er  96 hr • Tr anspi r at i on i s  r educed ( appr oxi mat el y  75%)  i n 100 and 1000 uM Cd by 24 hr  and i n 10 uM Cd by 96 hr • Tr anspi r at i on i s  compl et el y i nhi bi t ed by 1000 uM Cd af t er  96 hr • A mor e r api d decl i ne i n phot osynt hesi s r at e compar ed t o t r anspi r at i on,  suggest s t hat  f act or s ot her  t han st omat al  c l osur e ar e af f ect i ng phot osynt hesi s.  These coul d be due t o changes i n st r uct ur e and/ or  f unct i on of  phot osynt het i c pr ot ei ns Pr ot ei n Pr of i l e •   A new pr ot ei n ( 18 KDa)  appear s i n bean l eaves 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 • Appear ance of  t he 18 KDa pr ot ei n,  a l i kel y Cd st r ess r esponse pr ot ei n, i s  bot h t i me-  and concent r at i on- dependent • Dehydr i n pr ot ei n whi ch i s  a st r ess pr ot ei n and  pr ot ect s pl ant s agai nst dehydr at i on i s  st r ongl y pr esent  i n Cd t r eat ment s of 1000 uM at  48 hr  and of  10 uM at  and af t er  96 hr • Rubi sco and LHC- I I B pr ot ei n l evel s ar e not  af f ect ed by Cd;  however , t hei r  act i v i t y  mi ght  be • Cd af f ect s t hyl akoi d membr ane per meabi l i t y  whi ch may l ead t o a decl i ne i n pr ot on gr adi ent  acr oss t he membr ane and a r educt i on i n phot osynt hesi s r at eI  woul d l i ke t o t hank my Bi ol ogy 352 l ab par t ner s Br eanne Johnson,  Samant ha I ver sen and Rosanne Pi puni c  f or  hel p wi t h a pr el i mi nar y exper i ment . I  woul d al so l i ke t o t hank Shi ng Zhan f or  hel pi ng wi t h some dat a col l ect i on.  Fi nal l y,  I  woul d l i ke t o t hank Skyl i ght  Devel opment  Fund,  Facul t y of Sci ence and t he Depar t ment  of  Bot any f or f i nanci al  suppor t .  Fi gur e 1 - Ti me Cour se Changes i n  Mor phol ogy of  Cont r ol and Cd Tr eat ed bean pl ant s 6 hr 24 hr 48 hr 96 hr 168 hr 0 1 2 3 4 5 6 7 P h o to s y n th e s is  R a te  ( u m o l. m-2 s- 1 ) 0 24 48 96 168 Time (hr.) Control 10uM Cd 100 uM Cd 1000uM Cd 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 T ra n sp ir a ti on  R at e (u m ol .m -2  s ) 0 24 48 96 168 Time (hr.) Cont rol 10uM Cd 100 uM Cd 1000uM Cd Acknowl edgement s 66 KDa 45 KDa 36 KDa29 KDa 24 KDa 20 KDa 14. 2 KDa Fi gur e 4  -  Pol yacr y l ami de gel showi ng t he maj or  pr ot ei n bands i n 24 and 48 hr  cont r ol  and Cd t r eat ed bean pl ant s St andar d Pr ot ei n s 66 KDa 45 KDa 36 KDa 29 KDa 24 KDa 20 KDa 14. 2 KDa St andar d Pr ot ei n s Fi gur e 5  -  Pol yacr y l ami de gel showi ng t he maj or  pr ot ei n bands i n 24 and 48 hr  cont r ol  and Cd t r eat ed bean pl ant s Fi gur e 6 -  West er n Bl ot ,  Dehydr i n ( 60 KDa)   i n 4 and 6 hr  cont r ol and Cd t r eat ed bean pl ant s  Fi gur e 7 - West er n Bl ot , Dehydr i n ( 60 KDa)   i n 24 and 48 hr  cont r ol and Cd t r eat ed bean pl ant s Fi gur e 9 - West er n Bl ot  LHC- I I B ( 25 KDa)  i n 48 hr  cont r ol  and Cd t r eat ed bean pl ant s 0 hr  Cont r ol  10 uM  100 uM  1000 uM Cont r ol 10uM  100 uM  1000 uM  Cont r ol  10 uM 100 uM  1000 uM Cont r ol 10 uM  100 uM  1000 uM Fi gur e 2 -  Ti me Cour se Changes i n phot osynt hesi s Rat e ( umol . m- 2 s - 1 )  of   Cont r ol  and Cd Tr eat ed bean pl ant s Fi gur e 3 -  Ti me Cour se Changes i n Tr anspi 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   1.  Mor phol ogi cal  and physi ol ogi cal ef f ect s of  Cd ar e bot h t i me-  and concent r at i on- dependent . 2.  Phot osynt hesi s and t r anspi r at i on r at es ar e i nhi bi t ed by Cd 3.  A new pr ot ei n ( 18 KDa)  i s det ect ed i n r esponse t o Cd st r ess. Cont r ol  10 uM  100 uM 1000 uM 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 by heavy met al s. Phot osynt hesi s Resear ch.  7:  31- 40,  1985. 3.  Sandal i o,  L. M. ,  Dal ur zo,  H. C. ,  Gomez,  M. ,  Romer o- Puer t as,  M. C.  and del  Ri o,  L. A. 2001.  Jour nal  of  Exper i ment al  Bot any 52:  2115- 2126.   Envi r onment al  pol l ut i on has i ncr eased dr amat i cal l y  over  t he past f ew decades mai nl y because of  human act i v i t i es ( 1) .  Pr esence of  Cd causes di f f er ent  t ypes of  pr obl ems t o l i v i ng bei ngs i ncl udi ng a wel l - known cancer  causat i on i n human ( 2) . Si nce pl ant s as pr i mar y pr oducer s ar e consumed by ani mal s and human and al so because of  t he usage of pl ant s i n dr ug pr oduct i on,  i t  i s necessar y t o st udy t he ef f ect s of  Cd on pl ant s ( 1) .  Pr evi ous st udi es have shown negat i ve ef f ect s on mor phol ogy and r educed phot osynt hesi s and t r anspi r at i on r at e i n pr esence of  Cd ( 2) .  Thi s has been at t r i but ed t o t he i nt er f er ence of  t he el ement  wi t h mol ecul ar  pat hways such as phot osyst em I I  and el ect r on t r anspor t  chai n i n t hyl akoi d membr ane  ( 2) .  Al so,  st omat al  c l osur e i n Cd st r ess condi t i ons r esul t s i n l ower  t r anspi r at i on r at e and l ower CO2 i nt ake whi ch i n  t ur n  causes l ower phot osynt hesi s r at e ( 2) .  Cd i s al so known t o di s t ur b t he st r uct ur e and act i v i t i es of  enzymes and al t er  t he l i pi d composi t i on and per meabi l i t y of  membr ane ( 3) .  Fi nal l y accumul at i on of  Cd i n pl ant  or gans such as shoot  and r oot  may i nt r oduce i nhi bi t or s t o r egul at e pr ocesses of pl ant s such as ni t r at e assi mi l at i on pat hway ( 3) . The obj ect i ve of  t hi s  s t udy i s  t o exami ne t he ef f ect  of  cadmi um on mor phol ogy,  phot osynt hesi s , t r anspi r at i on,  and t he pr ot ei n pr of i l es. 24 hr Cont r o l  24 hr 10 uM 24 hr 100 uM 24 hr 1000 uM 48 hr Cont r o l 48 hr 10 uM 48 hr 100 uM 48 hr 1000 uM 96 hr Cont r o l 96 hr 10 uM 96 hr 100 uM 96 hr 1000 uM 168 hr Cont r o l 168 hr 10 uM Lar ge subuni t of  Rubi sco & ATPase (  54KDa) LHC-  I I B  ( 25 KDa) Newl y synt hesi sed pr ot ei n ( 18 KDa) Lar ge subuni t  of Rubi sco and ATPase      ( 54 KDa) LHC-  I I B ( 25 KDa) Newl y synt hesi sed pr ot ei n ( 18 KDa) Fi gur e 8 -  West er n Bl ot  Dehydr i n ( 60 KDa)  i n 96 and 168 hr cont r ol  and Cd t r eat ed bean pl ant s 96 hr 168 hr Cont r ol 10 uM 100 uM 1000 uM Cont r ol 10 uM 24 hr 48 hr 4 hr 6 hr Fut ur e St udi es •  I dent i f y  st r uct ur e and f unct i on of t he 18 KDa pr ot ei n •  St udy Cd ef f ect s at  t he membr ane per eabi l i t y  l evel •  I nvest i gat e Cd ef f ect s on gene expr essi on

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