Unique features of the rice blast resistance Pish locus revealed by large scale retrotransposon-tagging 英文参考文献.docVIP

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Unique features of the rice blast resistance Pish locus revealed by large scale retrotransposon-tagging 英文参考文献.doc

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Unique features of the rice blast resistance Pish locus revealed by large scale retrotransposon-tagging 英文参考文献

Takahashietal.BMCPlantBiology2010,10:175 /1471-2229/10/175 RESEARCH ARTICLE OpenAccess Uniquefeaturesofthericeblastresistance Pishlocusrevealedbylargescale retrotransposon-tagging AkiraTakahashi1,NagaoHayashi1,AkioMiyao2,HirohikoHirochika2* Abstract Background:Rgene-mediatedresistanceisoneofthemosteffectivemechanismsofimmunityagainstpathogens inplants.Todatesomecomponentsthatregulatetheprimarystepsofplantimmunityhavebeenisolated, however,themoleculardissectionofdefensesignalingdownstreamoftheRproteinsremainstobecompleted.In addition,Rgenesareknowntobehighlyvariable,however,themolecularmechanismsresponsibleforthis variabilityremainobscure. Results:ToidentifynovelfactorsrequiredforRgene-mediatedresistanceinrice,weusedriceinsertionalmutant lines,inducedbytheendogenousretrotransposonTos17,inageneticscreeninginvolvingthericeblastfungus Magnaportheoryzae.Weinoculated41,119mutantlineswiththefungususingahighthroughputprocedure,and identified86mutantlineswithdiminishedresistance.Agenomeanalysisrevealedthat72ofthe86linescontained mutationsinageneencodinganucleotidebindingsite(NBS)andleucinerichrepeat(LRR)domain-containing (NBS-LRR)protein.AgeneticcomplementationanalysisandapathogenesisassaydemonstratedthatthisNBS-LRR geneencodesPish,whichconfersresistanceagainstracesofM.oryzaecontainingavrPish.Theother14lineshave intactcopiesofthePishgene,suggestingthattheymaycontainmutationsinthesignalingcomponents downstreamofPish.ThegenomeanalysisindicatedthatPishanditsneighboringthreeNBS-LRRgenesarehigh similartooneanotherandaretandemlylocated.AninsilicoanalysisofaTos17flankingsequencedatabase revealedthatthisregionisa“hotspot”forinsertion.Intriguingly,theinsertionsitesarenotdistributedevenly amongthesefourNBS-LRRgenes,despitetheirsimilarityatthesequenceandexpressionlevels. Conclusions:InthisworkweisolatedtheRgenePish,andidentifiedseveralothermutantsinvolvedinthesignal transductionrequiredforPish-mediatedresistance.Theseresultsindicatethatourgeneticapproachisefficientand usefulforunveili