基于Ypt1基因为靶标的木香疫病病组织及病田土壤分子检测技术.pdfVIP

植物病理学报 ACTAPHYTOPATHOLOGICASINICA 43(1)：95—99(2013) MolecularDetection ofPhytophthora tentaculata in InfectedAucklandialappaTissuesand SoilontheBasisofYpttGene MENG Jun，LIU Yan，FANG Zhi—xiang (NationalKeyLaboratoryofBiosafety，NanjingInstituteofEnvironmentalSciences MinistryofEnvironmentalProtectionofChina，Nanjing210042，China) Abstract：A species．specificPCR assaywasestablished forrapidand accuratedetection oftheoomycete pathogen Phytophthoratentaculataindiseasedplanttissuesandinfectedsoil．A pairofspecies—specificprimers Ptl／Pt2weredesignedonthebasisofRasrelatedprotein(Ypt1)genesequencesofthePhytophthoraspecies． PCR amplificationwiththePtprimersresultedina386bpproductonlyfrom isolatesofP．tentaculata．The detection thresholdwith Ptprimerswas100 Pg ofgenomicDNA．A nested PCR procedurewasdeveloped usingYptlF／YptlR asthefirst—roundamplificationprimersandPt1／ ashtesecond—roundprimers．which increasedthedetectionsensitivitylO0．ofldtolpg．PCR usingthesePtprimerscanalsobeusedtodetectP． tentaculatainnaturallyinfectedplanttissuesandsoil．ThePCR—basedmehtoddevelopedinthisstudyprovides arapidandsensitivetoolofrdetectionofP．tentaculata． Keywords：moleculardetection； tlgene；oomycetes 基于 yp 基因为靶标的木香疫病病组织及病田土壤分子检测技术 孟 军 ，刘 燕，方志翔 (环境保护部南京环境科学研究所国家环境保护生物安全重点实验室，南京210042) 文章编号：0412—0914(2013)01-0095—05 In August2007，stalk rotsymptomswereob_ diseasescausedbyPhytophthora 1nfection and for servedonAucklandialappa (Asteraceae)，aneco— reducingpathogenspread． nomicallyimportantChinesemedicinalplant，inthe The study wasaimedatdevelopinga species— majorityoffieldsinYunnanProvinceofChina．This specificPCR assayofrdetectionofP．tentaculatain washte firstreportofPhytophthora tentaculata in infectedtissuesandsoilsamples．Theobjectivewas ChinaLlj to design a pair ofP． tentaculat