Overexpression of ORCA3 and G10H in Catharanthus roseus Plants Regulated Alkaloid Biosynthesis and Metabolism Revealed by NMR-Metabolomics 英文参考文献.docVIP

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Overexpression of ORCA3 and G10H in Catharanthus roseus Plants Regulated Alkaloid Biosynthesis and Metabolism Revealed by NMR-Metabolomics 英文参考文献.doc

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Overexpression of ORCA3 and G10H in Catharanthus roseus Plants Regulated Alkaloid Biosynthesis and Metabolism Revealed by NMR-Metabolomics 英文参考文献

OverexpressionofORCA3andG10HinCatharanthus roseusPlantsRegulatedAlkaloidBiosynthesisand MetabolismRevealedbyNMR-Metabolomics QifangPan1,2,QuanWang1,FangYuan1,ShihaiXing1,JingyaZhao1,YoungHaeChoi2, RobertVerpoorte2,YueshengTian1,GuofengWang1,KexuanTang1* 1PlantBiotechnologyResearchCenter,SJTU-CornellInstituteofSustainableAgricultureandBiotechnology,Fudan-SJTU-NottinghamPlantBiotechnologyRDCenter, SchoolofAgricultureandBiology,ShanghaiJiaoTongUniversity,Shanghai,People’sRepublicofChina,2NaturalProductsLaboratory,InstituteofBiologyLeiden,Leiden University,Leiden,TheNetherlands Abstract InordertoimprovetheproductionoftheanticancerdimericindolealkaloidsinCatharanthuseroseus,muchresearchhas been dedicated to culturing cell lines, hairy roots, and efforts to elucidate the regulation of the monoterpenoid indole alkaloid(MIA)biosynthesis.Inthisstudy,theORCA3(Octadecanoid-derivativeResponsiveCatharanthusAP2-domain)gene alone orintegrated withtheG10H(geraniol 10-hydroxylase)gene werefirstintroduced into C.roseus plants.Transgenic C.roseusplantsoverexpressingORCA3alone(ORlines),orco-overexpressingG10HandORCA3(GOlines)wereobtainedby geneticmodification.ORCA3overexpressioninducedanincreaseofAS,TDC,STRandD4Htranscriptsbutdidnotaffect CRMYC2andG10Htranscription.G10HtranscriptsshowedasignificantincreaseunderG10HandORCA3co-overexpression. ORCA3 and G10H overexpression significantly increased the accumulation of strictosidine, vindoline, catharanthine and ajmalicinebuthadlimitedeffectsonanhydrovinblastineandvinblastinelevels.NMR-basedmetabolomicsconfirmedthe higher accumulation of monomeric indole alkaloids in OR and GO lines. Multivariate data analysis of 1H NMR spectra showedchangeofaminoacid,organicacid,sugarandphenylpropanoidlevelsinbothORandGOlinescomparedtothe controls.TheresultindicatedthatenhancementofMIAbiosynthesisbyORCA3andG10Hoverexpressionmightaffectother metabolicpathwaysintheplantmetabolismofC.roseus. Citation:PanQ,WangQ,YuanF,XingS,ZhaoJ,etal.(2012)Overexpressionof