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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。
A pair of pine wood nematode-specific PCR primers (upstream primer: 5’-CTACGTGCTGTTGTTGAGTTGGC-3 ’, and downstream primer: 5’-TGGTGCCTAACATTGCGCGA-3’) screened from this study were used to amplify one The length of 403bp (GenBank accession number: DQ855275) of the specific DNA fragments. This primer can be used to bind pine wood nematode DNA to Monochamus alternatus, as well as to pine wood nematode, Bursaphelenchus xylophilus, Bursaphelenchus xylophilus, Bursaphelenchus xylophilus, Nematodes, Bladeslaederzia, Procyontera nematode, Steinerneri longipes nematode, Stem nematodes, Skeletal hookworm two stomach worms and parasitic rodworm nematodes and other 12 kinds of nematode DNA to distinguish. On this basis, a set of technology to detect pine wood nematode directly from the infected Monochamus alternatus was established by using PCR technique, and was tested in practical application.