东亚型和欧洲型拟松材线虫的形态学和ITSPCR-RFLP鉴定

Identification of East Asian and European Types of Bursaphelenchus mucronatus by Morphological and ITS PCR-RFLP Methods

对宁波口岸截获的不同来源木质包装中的东亚型和欧洲型拟松材线虫进行形态学和ITSPCR-RFLP研究。选用5种限制性内切酶RsaI,HaeⅢ,MspI,HinfI,AluI对各株系单条线虫ITS区PCR扩增产物进行酶切,发现限制性内切酶RsaI有3种酶切结果,表明ITS区段存在DNA序列异质性。研究再次证明ITSPCR-RFLP不但是伞滑刃线虫鉴定十分可靠的辅助手段,还可有效区分种内不同地理型。对拟松材线虫与松材线虫(包括"M"型和"R"型株系)、豆伞滑刃线虫、伪伞滑刃线虫等近似种的区别进行讨论。

Morphological and ITS PCR-RFLP methods were used to identify East Asian and European types of Bursaphelenchus mucronatus isolated in Ningbo, China from packaging wood imported from different countries. Five traditional restriction endonucleases： Rsa I, Hae Ⅲ , Msp l, Hinf I and Alu I were applied to digest the PCR products of ITS region from single nematode of different isolates. It is first reported that there are 3 different patterns after Rsa I digestion, which may be caused from DNA sequence heterogeneity in ITS region. ITS-RFLP method is an efficient ~thod for Bursaphelenchus identification, and can be used for analyzing intraspecific different geographic types. 3al characters of B. mucronatus compared with similar species, such as B. xylophilus （including ＂ R＂ and ）, B. doui and B. fraudulentus, were also discussed.