基于SCAR标记的小麦禾谷孢囊线虫快速分子检测技术

Rapid Molecular Diagnosis Based on SCAR Marker System for Cereal Cyst Nematode

【目的】建立从禾谷孢囊线虫及其近缘种的混合群体中检测禾谷孢囊线虫的快速分子检测方法。【方法】使用随机扩增多态性DNA(RAPD)和特征序列扩增区域(SCAR)标记的方法,运用PCR技术进行特异性检测。【结果】用本研究建立的SCAR标记快速分子检测体系对9种32个线虫种群进行检测,能够直接从混合线虫样品中检测出禾谷孢囊线虫。该检测方法对禾谷孢囊线虫的孢囊、2龄幼虫具有扩增能力,最低检出阈值为1/2000个孢囊,1/80头2龄幼虫。【结论】本研究设计的SCAR标记快速分子检测体系能够从禾谷孢囊线虫及其近缘种的混合种群中快速检测出禾谷孢囊线虫,且检测准确、灵敏度高。

【Objective】 The objective of this study is to build a SCAR marker system for rapid molecular detection of cereal cyst nematode （Heterodera avenae） from mixtures of different nematodes. 【Method】 With the method of RAPD and SCAR, a single-step PCR was used to specially detect H. avenae. 【Result】 Using the SCAR marker system established in this study, the target fragment was amplified from the DNA templates of H. avenae only, from other 8 species including 32 populations. The SCAR marker system could detect cysts and the J2 of H. avenae, and the specific fragment could be clearly identified when the dilution was 1/2000 of a cyst or 1/80 of a J2 for all replicates. 【Conclusion】 The establishment of the SCAR marker system for rapid molecular detection of H. avenae can be used to detect the H. avenae from the mixed nematode samples rapidly, and the detection is accurate and highly sensitive.