Triacylglycerol synthesis by PDAT1 in the absence of DGAT1 activity is dependent on re-acylation of LPC by LPCAT2 英文参考文献.docVIP

Triacylglycerol synthesis by PDAT1 in the absence of DGAT1 activity is dependent on re-acylation of LPC by LPCAT2 英文参考文献.doc

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Triacylglycerol synthesis by PDAT1 in the absence of DGAT1 activity is dependent on re-acylation of LPC by LPCAT2 英文参考文献

Xuetal.BMCPlantBiology2012,12:4 /1471-2229/12/4 RESEARCH ARTICLE OpenAccess TriacylglycerolsynthesisbyPDAT1intheabsence ofDGAT1activityisdependentonre-acylationof LPCbyLPCAT2 JingyuXu1,AndersSCarlsson2,TammyFrancis1,MengZhang3,TravisHoffman1,MichaelEGiblin1and DavidCTaylor1,4* Abstract Background:TheArabidopsisthalianadgat1mutant,AS11,hasanoilcontentwhichisdecreasedby30%,anda stronglyincreasedratioof18:3/20:1,comparedtowildtype.DespitelackingafunctionalDGAT1,AS11stillmanages tomake70%ofWTseedoillevels.Recently,itwasdemonstratedthatintheabsenceofDGAT1,PDAT1was essentialfornormalseeddevelopment,andisadominantdeterminantinArabidopsisTAGbiosynthesis. Methods:Biochemical,metabolicandgeneexpressionstudiescombinedwithgeneticcrossingofselected ArabidopsismutantshavebeencarriedouttodemonstratethecontributionofArabidopsisPDAT1andLPCAT2in theabsenceofDGAT1activity. Results:ThroughmicroarrayandRT-PCRgeneexpressionanalysesofAS11vs.WTmid-developingsiliques,we observedconsistenttrendsbetweenthetwomethods.FAD2andFAD3wereup-regulatedandFAE1down- regulated,consistentwiththeAS11acylphenotype.PDAT1expressionwasup-regulatedbyca65%whilePDAT2 expressionwasup-regulatedonly15%,reinforcingthedominantroleofPDAT1inAS11TAGbiosynthesis.The expressionofLPCAT2wasup-regulatedby50-75%,whileLPCAT1expressionwasnotsignificantlyaffected.Invitro LPCATactivitywasenhancedby75-125%inmicrosomalproteinpreparationsfrommid-developingAS11seedvs WT.Co-incidenthomozygousknockoutlinesofdgat1/lpcat2exhibitedseverepenaltiesonTAGbiosynthesis, delayedplantdevelopmentandseedset,evenwithafunctionalPDAT1;thedoublemutantdgat1/lpcat1showed onlymarginallyloweroilcontentthanAS11. Conclusions:Collectively,thedatastronglysupportthatinAS11itisLPCAT2up-regulationwhichisprimarily responsibleforassistinginPDAT1-catalyzedTAGbiosynthesis,maintainingasupplyofPCasco-substrateto transfersn-2moietiestothesn-3positionoftheenlargedAS11DAGpool. Keywords:dgat1mutantAS11,LPCAT1,LPCAT2,PDAT1,OilbiosynthesisSeedlinesfromNottinghamArabidopsis

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