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林业科学 ›› 2005, Vol. 41 ›› Issue (6): 101-104.doi: 10.11707/j.1001-7488.20050616

• 论文及研究报告 • 上一篇    下一篇

疫木切面上松材线虫抗原的免疫学检测方法

白钢1 李海燕2 马洪周1 王玉嬿2 杨文博1 张奇1   

  1. 1.南开大学生命科学学院,天津300071;2.国家林业局森林病虫害防治总站,沈阳110034;
  • 收稿日期:2003-12-12 修回日期:1900-01-01 出版日期:2005-11-25 发布日期:2005-11-25

An Immunocytochemical Method for Identification of Bursaphelenchus xylophilus Antigen on the Surface of Pinewood Section

Bai Gang1,Li Haiyan2,Ma Hongzhou1,Wang Yuyan2,Yang Wenbo1,Zhang Qi1   

  1. 1.College of Life Science, Nankai University Tianjin 300071; 2.General Station of Forest Pest Management,State Forestry Administration Shenyang 110034
  • Received:2003-12-12 Revised:1900-01-01 Online:2005-11-25 Published:2005-11-25

摘要:

利用抗松材线虫血清建立在疫木切面上直接进行免疫组化染色检测松材线虫抗原的方法。ELISA和免疫斑点印迹的结果表明:该方法对松材线虫具有较好的特异性,可以检测出木材表面0.1μg的线虫抗原,显示了较好灵敏度。自然感染松材线虫的黑松疫木免疫组化的染色结果表明:该方法可以清晰地检出管胞处松材线虫蛋白抗原的存在。

关键词: 松材线虫, 线虫分析, 免疫组化, ELISA

Abstract:

There are three different views regarding pathogenic mechanism of pinewood nematode disease: enzymes, cavitation, toxins.Most conclusions are based upon the change of the morphology of infected wood, but fail in expounding the internal mechanism on the molecular level. A immuno-cytochemical method for detecting BX (Bursaphelenchus xylophilus)antigen in pinewood section was established with anti-BX serum. The method not only showed a new potential for the direct detection of BX on pine wood section, but also discovered yield information about BX and nematode solvable antigen in wood tissues, and could be further confirmed synthetic matrices on pathogen distribution. Combining aspects of ELISA and immunoblot, the method was specific to nematode antigen two species of Bursaphelenchus genus, and could identify 0.1 μg nematode protein on the surface of pinewood section. With this method, it was detected that the BX antigen in tracheid tissue of Pinus thunbergii which were representative of a natural infestation.

Key words: Bursaphelenchus xylophilus, nematode analysis, immunocytochemistry, ELISA