安徽桑黄花型萎缩病植原体16S rDNA序列分析及分子检测.pdfVIP

植物病理学报 ACTAPHYTOPATHOLOGICASINICA 40(6)：642Jo46(2010) 安徽桑黄花型萎缩病植原体 16S rDNA 序列分析及分子检测 卢全有 (中国农业科学院蚕业研究所，农业部蚕桑遗传改 良重点开放实验室，镇江 212018；江苏科技大学 ，镇江 212003) Moleculardetectionandsequenceanalysison16S ribosomalDNA ofphytoplasma associatedw ith mulberry yellow dwarfdisease occurring in Anhui LU Quan．you (TheKeyLaboratoryofGeneticImprovementofSilkworm andMulberry，MinistryofAgriculture，TheSericultrualResearch Institute，ChineseAcademyofAgriculturalSciences，Zhenjiang212018，China；JinagsuUniversityofScienceandTechnology， Zhenjiang212003，China) Abstract：Mulberryyellowdwarf (MYD)diseaseisanquarantinediseaseandthecausalagentisaphytoplas— ma．Twopairsofpublisheduniversalprimer，P1／P7andRm 16F2／Rm16R1，basedonthe16S一23SrDNA se— quenceofphytoplasmaandtotalDNA extractedrfom infected mulberrytissueswereemployedforPCR and nested—PCR detection．Theresultsrevealedthataphytoplasma ·specificl830bprfagmentwithaG +C con· tentof46．01％ wassequenced(GenBankaccessionNo．GQ249410)．Thesequenceshared99．7％ and 99．8％ identitywithasteryellows．therepresentatiivephytoplasmain16SrIgroup．andmulberrydwarfphyto— plasmaclassifiedintosubgroupBin16SrigroupandnamedastheMYDphytoplasmastrainAnhui(MYD— Anh)．Aphylogenetictreebasedon16SrDNA sequenceswasconstructedandshowedthatMYD—Anhwas clusteredinto16SrIgroup．Identity of16SrDNA sequencebetweenMYD—Anh andmulberry yellow dwarf phytoplasmastrainZhenjiang(MD—zj)wasnearly100％，andtheymightbelongtothesamestrain．Nested— PCR wasused to detectthepathogenicphytoplasmafrom thedifferentialtissuesofmulberry infectedwiht MYD—Anh．Theresultsshowedthataphytoplasma—specific1．4kbrfagmentwasamplifiedwiht totalDNA ex- tractedrfom barkandvein．Nested—PCR wasmoresensitivethanPCR ofrdetectingMYD phytoplasma． Keywords：mulberryyellow dwarfphytoplasma；PCR；phylogenetic；nested—PCR 文章编号：0412—0914(2010)06-0642435 桑黄花型萎缩病 (Mulberryyellow dwrafdi— 人工培养，根据形态和培养性状无法对其分类和鉴 sease．MYD)是桑树上的主要