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香蕉穿孔线虫(Radopholus similis)是一种国际公认的极具毁灭性的有害线虫,也是我国重要的进境检疫性有害生物。利用线虫通用引物对香蕉穿孔线虫7个不同群体的ITS进行PCR扩增、克隆及测序,获得ITS片段序列长度为706bp。经与国外香蕉穿孔线虫种群及近缘种的ITS序列进行比对及同源性分析,构建设计了1对香蕉穿孔线虫的特异性引物RsF1/RsR1,特异性扩增片段长度为271 bp;同时引入D2A/D3B作内标,研究出特异性检测香蕉穿孔线虫的一步双重PCR分子技术和方法,该项检测技术特异性好,耗时较短、操作简便、可靠。
Radopholus similis is an internationally recognized devastating harmful nematode and is also an important quarantine pest in our country. The ITS sequences of seven different populations of B. perfringens were amplified by PCR, cloned and sequenced using the universal nematode primers. The ITS sequence length was 706 bp. A pair of RsF1 / RsR1 specific primers were designed and constructed based on the ITS sequences of the foreign banana population and its relative species. The length of the specific amplified fragment was 271 bp. At the same time The introduction of D2A / D3B as an internal standard, developed a one-step double PCR molecular techniques and methods for the specific detection of banana boreal nematodes, the detection technology is specific, short time-consuming, easy to operate, and reliable.