Functional Significance of AtHMA4 C-Terminal Domain In Planta 英文参考文献.docVIP

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Functional Significance of AtHMA4 C-Terminal Domain In Planta 英文参考文献.doc

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Functional Significance of AtHMA4 C-Terminal Domain In Planta 英文参考文献

Functional Significance ofAtHMA4 C-Terminal Domain InPlanta RebeccaF.Mills1,BillyValdes1,MichaelDuke1,KerryA.Peaston1,BrettLahner2,DavidE.Salt2, LorraineE.Williams1* 1School of Biological Sciences, University of Southampton, Southampton, Hampshire, United Kingdom, 2Center for Plant Environmental Stress Physiology, Purdue University,WestLafayette,Illinois,UnitedStatesofAmerica Abstract Background: Enhancing the upward translocation of heavy metals such as Zn from root to shoot through genetic engineering has potential for biofortification and phytoremediation. This study examined the contribution of the heavy metal-transportingATPase,AtHMA4,totheshootionomicprofileofsoil-grownplants,andinvestigatedtheimportanceof theC-terminaldomaininthefunctioningofthistransporter. PrincipalFindings:TheArabidopsishma2hma4mutanthasastuntedphenotypeandadistinctiveionomicprofile,withlow shootlevelsofZn,Cd,Co,KandRb,andhighshootCu.ExpressionofAtHMA4(AtHMA4-FL)undertheCaMV-35Spromoter partiallyrescuedthestuntedphenotypeofhma2hma4;rosettediameterreturnedtowild-typelevelsinthemajorityoflines and bolts were also produced, although the average bolt height was not restored completely. AtHMA4-FL expression rescued Co, K, Rb and Cu to wild-type levels, and partially returned Cd and Zn levels (83% and 28% of wild type respectively).Incontrast,expressionofAtHMA4-trunc(withouttheC-terminalregion)inhma2hma4onlypartiallyrestored therosettediameterintwooffivelinesandboltproductionwasnotrescued.Therewasnosignificanteffectontheshoot ionomic profile, apart from Cd, which was increased to 41% of wild-type levels. When the AtHMA4 C-terminal domain (AtHMA4-C-term) was expressed in hma2 hma4 it had no marked effect. When expressed in yeast, AtHMA4-C-term and AtHMA4-truncconferredgreaterCdandZntolerancethanAtHMA4-FL. Conclusion:Theionomeofthehma2hma4mutantdiffersmarkedlyfromwtplants.Thefunctionalrelevanceofdomainsof AtHMA4inplantacanbeexploredbycomplementingthismutant.AtHMA4-FLismoreeffectiveinrestorings