《Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted NBS-LRR Protein Class Gene in Apple （Golden Delicious）》.pdfVIP

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《Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted NBS-LRR Protein Class Gene in Apple （Golden Delicious）》.pdf

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《Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted NBS-LRR Protein Class Gene in Apple （Golden Delicious）》.pdf

MolecularPlant ·Volume7 ·Number1 ·Pages218-230 ·January2014 RESEARCHARTICLE CloningandCharacterizationofmiRNAsand TheirTargets，IncludingaNovelmiRNA—Targeted NBS-LRRProteinClassGeneinApple(Golden Delicious) ChaoMa。一，YouLu--，SonglinBai。，WennanZhang。，XuweiDuan。，DongMeng。，ZhigangWang。一， AideWang，‘ZongshanZhou andTianzhongLi。’ aLaboratoryofFruitCellandMolecularBreeding，ChinaAgriculturalUniversity,Beijing100193，China bResearchInstituteofPomology,ChineseAcademyofAgriculturalSciences，Xingcheng，Liaoning125100，China cCollegeofHorticulture，ShenyangAgriculturalUniversity,Shenyang110866，China ABSTRACT MicroRNA (miRNA)hasemergedasanimportantregulatorofgeneexpressioninplants．146miRNAs wereIdentifiedfromapple(Malusdomesticacv,GoldenDelicious)bybioinformaticanalysisandRNA Iibrarysequenc- ing．From these，135wereconservedand11werenoveImiRNAs．TargetanalysispredictedoneofthenoveImiRNAs， Md．miRLn11(MalusdomesticamicroRNA Ln11)，targetedanapplenucleotide．bindingsite(NBS)一leucine．richrepeat (LRR)classproteincodinggene(Md．NBS)．5RACEassayconfirmedtheabilityofMd．miRLn11tocleaveMd．NBSatthe 11-12-ntposition．AnalysisoftheexpressionofMd-miRLnl1andMd-NBSduringtheoptimum invasionperiodin40 applevarietiesshowedthattheexpressionof，Wd．～B5geneinresistantvarietiesishigherthaninsusceptiblevarieties． withan inversepatternforMd-miRLn11．Seedlingsfrom theresistantapplevariety JiGuan wereusedtocarryout anAgrobacterium infiltrationassav-andthen inoculatedwiththeappleleafspotdisease．Theresultshowedaclear declineofdiseaseresistanceinJiGuanapples．1ncontrast．thesusceptiblevariety FujiinfiltratedwiththeM d-NBS geneshowedasignificantincreaseindiseaseresistance．Basedontheaboveresults，weproposethatMd-miRLn11 regulatesM d·NBSgeneexpressioninparticularundertheconditionofpathogeninfection．andthattheMd．miRLn11 targetingp-loopsitemayregulatemanyNBS-LRRproteinclassgenesinw oodyplants． Keywords： apple；microRNA；NBS—LRRprotein；bioinformaticsanalysis；genefunctio