旱稻孢囊线虫的快速分子检测

根据旱稻孢囊线虫(Heterodera elachista)和常见孢囊线虫的ITS序列比对分析,设计1对旱稻孢囊线虫特异性引物He–F/He–R,特异性片段长度为281 bp。运用该特异性引物及建立的DNA提取方法和PCR体系,可特异性检测旱稻孢囊线虫单条2龄幼虫,可以从混合有1条旱稻孢囊线虫的0.1 g水稻根组织中特异性检测出目的DNA片段。特异性引物He–F/He–R与通用引物D2A/D3B结合,运用一步双重PCR检测方法可快速鉴定单孢囊,也可从初始分离的田间土壤总线虫样品中直接检测出旱稻孢囊线虫。

Rapid molecular diagnosis of rice cyst nematode (Heterodera elachista)

Based on known ITS sequences of the rice cyst Nematode (Heterodera elachista) and common cyst nematodes, one species-specific primer pair named HeF and HeR was designed for H. Elachista, the expected fragment length of the PCR products using the primers was 281 bp. Using the established DNA extraction method and PCR system with the specific primers, target fragment detected from a single second instar larvae of nematode or an individual H. elachista mixed in 0.1 g rice root tissues. Combined the species-specific primers He-F and He-R with universal primer pairs D2A and D3B, a duplex PCR was established, which detects H. elachista from cyst nematode samples primarily separated from field soil.