【摘要】 利用本研究筛选出的一对松材线虫特异性PCR引物(上游引物:5′-CTACGTGCTGTTGTTGAGTTGGC-3′,下游引物:5′-TGGTGCCTAACATTGCGCGA-3′),从松材线虫DNA中,扩增出一条长度403bp(基因库登陆号:DQ855275)的特异性DNA片段。该引物可以将松材线虫DNA与松墨天牛组织以及拟松材线虫、畸刺伞滑刃线虫、变异伞滑刃线虫、小角伞滑刃线虫、利昂伞滑刃线虫、湖南伞滑刃线虫、红松滑刃线虫、滑刃属线虫、斯坦纳长尾线虫、小茎线虫、剑尾齿杆双胃线虫和寄生小杆属线虫等12种线虫的DNA区分开来。在此基础上,建立一套利用PCR技术直接从受感染的松墨天牛组织中检测出松材线虫的技术,并在实际运用中得到检验。更多还原

【Abstract】 The pine wood nematode(Bursaphelenchus xylophilus)is a highly damaging pest on a world scale.Monochamus alternatus is the most important vector of B.xylophilus.A quick,reliable and replicable method is needed for detecting the pest.To that end,we firstly developed a simple procedure for isolating DNA from the mixture of nematode +beetle tissue for subsequent nematode detection by PCR amplification.Then one pair of B.xylophilus-specific primers that generated a specific amplicon of 403 bp(DQ855275)located respectively in the ITS1 and ITS2 regions,were choosen.No amplicon was obtained from either M.alternatus or any of the 12 other nematodes species related to pine wilt disease(B.mucronatus,B.aberrans,B.corneolus,B.leoni,B.hunanensis,B.teratospicularis,Aphelenchoides resinosi,Seinura steineri,Ditylenchus parvus,Odorhabdiplogaster xiphocaudatus,Rhabditida sp.and Parasitorhabditis sp.)with the primers.The same 403 bp amplicon was also firstly obtained by PCR amplification from the beetle tissue infested with B.xylophilus.更多还原