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以锥尾伞滑刃线虫(Bursaphelenchus conicaudatus)作为对照,在韩国不同地域和寄主中采集了15个松材线虫分离物、3个拟松材线虫的分离物及5个未鉴定的该属的分离物,并对其进行了ITS和D2D3 rDNA序列分析。以单条雌线虫的DNA为模板,ITS和D2D3区由PCR仪扩增并克隆其序列。结果表明,所有松材线虫的ITS和D2D3区序列相同,没有种内变异。然而2个采自黑松和红松的拟松材线虫分离物的基因型分别是亚洲型和欧洲型。5个未鉴定线虫的数据表明,它们分别接近B.tusciae、B.lini、B.thailandae、B.doui和B.hylobianum,该结果同时被形态学结果所支持。利用5种酶(Hinf I、Alu I、Msp I、Hae III、Rsa),PCR产物克隆并测序的数据也可以区分不同种和基因型。
Using Bursaphelenchus conicaudatus as a control, 15 isolates of Bursaphelenchus xylophilus, 3 isolates of Bursaphelenchus xylophilus, and 5 unidentified isolates of this genus were collected from different regions and hosts in Korea The ITS and D2D3 rDNA sequences were analyzed. Using the DNA of a single female nematode as a template, ITS and D2D3 regions were amplified by a PCR machine and their sequences were cloned. The results showed that ITS and D2D3 sequences of all the Bursaphelenchus xylophilus were the same, with no intraspecies variation. However, the genotypes of two isolates of pine wood nematode collected from Pinus thunbergii and Pinus koraiensis were Asian and European, respectively. Data from five unidentified nematodes showed that they were close to B.tusciae, B.lini, B.thailandae, B.doui, and B.hylobianum, respectively, supported by morphological results. Data from the cloning and sequencing of the PCR products using five enzymes (Hinf I, Alu I, Msp I, Hae III, Rsa) also make it possible to distinguish between different species and genotypes.