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Effects of Cadmium on Morphology, Photosynthesis and Protein Profile of Bean Plants Pournia, Farnaz; Mehroke, Jarnail; Singh, Santokh 2009-03-14

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POSTER TEMPLATE BY:www.PosterPresentations.comEffects of Cadmium on Morphology, Photosynthesis and Protein Profile of Bean PlantsFarnaz Pournia; Jarnail Mehroke; Santokh SinghDepartment of Botany, Biology Program, University of British Columbia, Vancouver, B.C, CanadaAbstract.Figures Results and DiscussionMaterials and MethodsIntroductionConclusionsReferencesCadmium (Cd), a cancer-causing heavy metal ion,concentration in the environment has recently shown a rapidincrease mainly as a result of human activity. Since manyplants are used as medicine and food, it is crucial toinvestigate effects of Cd on plant growth and development.This study examines the time course and concentration-dependent effects of Cd on morphology, photosynthesis andprotein profiles in hydroponically-grown bean plants(Phaseolus vulgaris). High concentrations of Cd (1000 and100 uM) induced leaf wilting and reduced both leaf growthand photosynthesis rate within 24 and 48 hours respectively.Lower concentration of 10 uM Cd showed the wiltedmorphology after 96 hours. In addition, the Cd-inducedchanges in protein profiles especially the photosynthesis andstress proteins will be discussed in relation to morphology,leaf growth and photosynthesis in bean plants.Green Beans (Phaseolus vulgaris) were grown in thegreenhouse for 7 days. The seedlings were then transferredonto a hydroponic system which is a growing medium withknown concentrations of nutrients. After 7 more days (16 hr.light / day), the plants were treated with Cd at concentrationsof 10, 100 and 1000 uM.Growth and development of plants were monitored for sevendays. Rates of photosynthesis and transpiration for eachtreatment were measured at the end of 4, 6, 24, 48, 96 and168 hours after treatment using LI-6200 PortablePhotosynthetic System. By the end of each specified period,leaves from control and the three treatments were collectedfor protein [Ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco), Light Harvesting Complex-IIB (LHC-IIB) and Dehydrin)] analysis by SDS-PAGE and Western blottechniques.Morphology•High concentrations of Cd, 1000 and 100 uM inhibit growthand development of bean plants and cause wiltedmorphology in 24 hours and 48 hours respectively•The effects of low Cd concentration (10 uM) are observedafter 96 hr•Cd effects are both time- and concentration-dependent.Photosynthesis and Transpiration Rates•Photosynthesis is partially (approximately 50%) reduced inhigher Cd concentrations, 100 and 1000 uM, after 24 hr• Cd completely inhibited photosynthesis rate at and after 96hr•Transpiration is reduced (approximately 75%) in 100 and1000 uM Cd by 24 hr and in 10 uM Cd by 96 hr•Transpiration is completely inhibited by 1000 uM Cd after96 hr•A more rapid decline in photosynthesis rate compared totranspiration, suggests that factors other than stomatalclosure are affecting photosynthesis. These could be due tochanges in structure and/or function of photosyntheticproteinsProtein Profile• A new protein (18 KDa) appears in bean leaves treatedwith 1000 uM Cd at and after 48 hr, in 100 uM at and after96 hr and in 10 uM at 168 hr•Appearance of the 18 KDa protein, a likely Cd stressresponse protein, is both time- and concentration-dependent•Dehydrin protein which is a stress protein and protectsplants against dehydration is strongly present in Cdtreatments of1000 uM at 48 hr and of 10 uM at and after 96hr•Rubisco and LHC-IIB protein levels are not affected by Cd;however, their activity might be•Cd affects thylakoid membrane permeability which maylead to a decline in proton gradient across the membraneand a reduction in photosynthesis rateI would like to thank my Biology 352 lab partners Breanne Johnson, SamanthaIversen and Rosanne Pipunic for help with a preliminary experiment. I wouldalso like to thank Shing Zhan for helping with some data collection. Finally, Iwould like to thank Skylight Development Fund, Faculty of Science and theDepartment of Botany for financial support.Figure 1 -Time Course Changes in  Morphology of Control and Cd Treated bean plants 6 hr 24 hr 48 hr 96 hr 168 hr01234567Photosynthesis Rate (umol.m-2s-1 )0 24 48 96 168Time (hr.)Control10uM Cd100 uM Cd1000uM Cd00.20.40.60.811.21.41.61.8Transpiration Rate (umol.m-2 s) 0 24 48 96 168Time (hr.)Control10uM Cd100 uM Cd1000uM CdAcknowledgements66 KDa45 KDa36 KDa29 KDa24 KDa20 KDa14.2 KDaFigure 4 - Polyacrylamide gel showing the majorprotein bands in 24 and 48 hr control and Cd treatedbean plantsStandard Proteins66 KDa45 KDa36 KDa29 KDa24 KDa20 KDa14.2 KDaStandard ProteinsFigure 5 - Polyacrylamide gel showing the majorprotein bands in 24 and 48 hr control and Cd treatedbean plantsFigure 6 - Western Blot, Dehydrin (60 KDa)  in 4 and 6 hr control and Cd treated bean plantsFigure 7 -Western Blot,Dehydrin (60 KDa)  in 24 and 48 hr control and Cd treated bean plantsFigure 9 -Western Blot LHC-IIB (25 KDa) in 48 hr control and Cd treated bean plants0 hr Control 10 uM 100 uM 1000 uM Control 10uM 100 uM 1000 uMControl 10 uM 100 uM 1000 uM Control 10 uM 100 uM 1000 uMFigure 2 - Time Course Changes in photosynthesis Rate (umol.m-2 s-1 ) of  Control and Cd Treated bean plantsFigure 3 - Time Course Changes in Transpiration Rate    ( umol.m-2 s-1 ) of Control and Cd treated bean plants 1. Morphological and physiological effects of Cd are bothtime- and concentration-dependent.2. Photosynthesis and transpiration rates are inhibited by Cd3. A new protein (18 KDa) is detected in response to Cdstress.Control 10 uM 100 uM 1000 uM 1. Rai V.,Khatoon S., Bisht S. S. and Mehrotra S. 2005. Effects of Cadmium on growth, ultramorphology of leaf and secondarymetabolism of Phyllanthus amarus and Thorn. Chemosphere. 61 :1644-16502. Clijsters H., Van Assche F. 1985 Inhibition of photosynthesis by heavy metals. Photosynthesis Research. 7: 31-40, 1985.3. Sandalio, L.M., Dalurzo, H.C., Gomez, M., Romero-Puertas, M.C. and del Rio, L.A. 2001. Journal of Experimental Botany 52:2115-2126.Environmental pollution has increased dramatically over thepast few decades mainly because of human activities (1).Presence of Cd causes different types of problems to livingbeings including a well-known cancer causation in human(2). Since plants as primary producers are consumed byanimals and human and also because of the usage of plantsin drug production, it is necessary to study the effects of Cdon plants (1). Previous studies have shown negative effectson morphology and reduced photosynthesis andtranspiration rate in presence of Cd (2). This has beenattributed to the interference of the element with molecularpathways such as photosystem II and electron transportchain in thylakoid membrane (2). Also, stomatal closure inCd stress conditions results in lower transpiration rate andlower CO2 intake which in turn causes lower photosynthesisrate (2). Cd is also known to disturb the structure andactivities of enzymes and alter the lipid composition andpermeability of membrane (3). Finally accumulation of Cd inplant organs such as shoot and root may introduce inhibitorsto regulate processes of plants such as nitrate assimilationpathway (3).The objective of this study is to examine the effect ofcadmium on morphology, photosynthesis, transpiration, andthe protein profiles.24 hr Control24 hr10 uM 24 hr 100 uM 24 hr1000 uM48 hrControl48 hr10 uM48 hr100 uM48 hr1000 uM96 hr Control96 hr10 uM96 hr100 uM96 hr1000 uM168 hrControl168 hr10 uMLarge subunit of Rubisco & ATPase  ( 54KDa)LHC- IIB(25 KDa)Newly synthesised protein (18 KDa)Large subunit of Rubisco and ATPase           (54 KDa)LHC- IIB(25 KDa)Newly synthesised protein (18 KDa)Figure 8 - Western Blot Dehydrin (60 KDa) in 96 and 168 hr control and Cd treated bean plants96 hr 168 hrControl 10 uM 100 uM 1000 uM Control 10 uM24 hr 48 hr4 hr 6 hrFuture Studies• Identify structure and function of the 18 KDa protein• Study Cd effects at the membrane permeability level• Investigate Cd effects on gene expression

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