甘薯nbs-lrr类抗病基因同源序列的克隆及分析-cloning and analysis of homologous sequences of nbs - lrr disease resistance genes in sweet potato.docxVIP

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甘薯nbs-lrr类抗病基因同源序列的克隆及分析-cloning and analysis of homologous sequences of nbs - lrr disease resistance genes in sweet potato.docx

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甘薯nbs-lrr类抗病基因同源序列的克隆及分析-cloning and analysis of homologous sequences of nbs - lrr disease resistance genes in sweet potato

IsolationandCharacterizationofNBS-LRRtypeResistanceGeneAnalogsfromIpomoeabatatasAbstractSweetPotato(Ipomoea.batatas)wasimportantfoodcropwhichgrownwidelyaroundtheworld.ThemaindiseaseswhichthreattheproductionofsweetpotatowereStemnematode,Blackrot,BacterialWilt,Rootrotandsoon.Useingconventionalmethodstoattainthepotatoresistantvarietiesneedalongbreedingcycleandithasalowefficiency.Useingmolecularbiologymethodtoclonetheresisitancegenethenuseingthemethodofgeneticengineeringtobreednovelsweetpotatoresistantvarietiesmaybethemosteffectiveapproachtogetthehighresistantvarieties.Untilnowtherehavenoreportaboutthecloneofthesweetpotatoresistancegene.TheNBS-LRRtyperesisitancegenewerethemostcommonpopulationofRgenes,ithasbeenreportedthatplantdiseaseresistanceresponseandNBS-LRRproteinaredirectlyrelated.TheNBSdomainhavethefunctioninPi-bindingofATPorGTP,LRRdomainhavethefunctioninrecognitionpathogen.Thepolymerasechainreaction(PCR)wasperfomedforamplifingRGAsbyusingtheprimerswhichweredesignedaccordingtotheP-LOOPandGLPLAmotifoftheNBS-LRRprotein,andthegenomicDNAextractedfromAB94078-1,Xushu18andtheir8progenieswereutilizedastemplates.Atlast,26resistance-geneanalogsfromsweetpotatowereobtainedandallwereacceptedbyNCBI.These26RGAsweresortedintotwodistincttypeintermsoftheirputativeaminoacidsquencess,TIR-NBS-LRRtypeandnon-TIR-NBS-LRRtype,soitconfirmedtheearlyreportthatNBS-LRRgenehastwomajorgroupsindicotspecies.Furtheranalysisaboutthe26RGAaminoacidstructuresrevealthatthefollowing5sequencesR1100-AB,R1100-16,R1100-34,R1100-79,R1100-169inaditiontohavefourconservativemotifoftheNBSdomain,itsC-terminalaminoacidsequenceexistLRRstructure,whichhasnotbeenreportedbeforeourresearch.TheseRGAwereperformedclusteranalysiswiththepublishedsweetpotatoNBS-LRRRGAinGenBankdatabase,theresultshowedthattheRGAclonefromdifferentsweetpotatovarietiesbelongtodifferentgroups,theRGAinourreaserchcouldclassifiedastwonewcategories,R510andR1100groups.FromanotheraspecttheRGAmayprovidesomeevidenceaboutplantpopulationdifferentiation.Thesout