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林业科学 ›› 2012, Vol. 48 ›› Issue (8): 113-117.doi: 10.11707/j.1001-7488.20120818

• 论文 • 上一篇    下一篇

东亚型和欧洲型拟松材线虫的形态学和ITS PCR-RFLP鉴定

王江岭, 顾建锋, 陈先锋, 段维军   

  1. 宁波出入境检验检疫局 宁波 315012
  • 收稿日期:2011-02-11 修回日期:2012-01-23 出版日期:2012-08-25 发布日期:2012-08-25
  • 通讯作者: 顾建锋

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

Wang Jiangling, Gu Jianfeng, Chen Xianfeng, Duan Weijun   

  1. Ningbo Entry-Exit Inspection and Quarantine Bureau Ningbo 315012
  • Received:2011-02-11 Revised:2012-01-23 Online:2012-08-25 Published:2012-08-25

摘要:

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ITS PCR-RFLP研究。选用5种限制性内切酶Rsa I,Hae Ⅲ,Msp I,Hinf I,Alu I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Rsa I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ITS PCR-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。

关键词: 拟松材线虫, 鉴定, ITS-RFLP, 异质性

Abstract:

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 I, 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 assistant method for Bursaphelenchus identification, and can be used for analyzing intraspecific different geographic types. Morphological characters of B. mucronatus compared with similar species, such as B. xylophilus (including "R" and "M" forms), B. doui and B. fraudulentus, were also discussed.

Key words: Bursaphelenchus mucronatus, identification, ITS PCR-RFLP, heterogeneity

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