螺旋粉虱的分子检测方法

Molecular detection of Aleurodicus disperses(Homoptera: Aleyrodidae)

螺旋粉虱(Aleurodicus dispersus)是一种世界性重要入侵害虫。针对其体型微小难以进行准确、快速鉴别,以及对其卵、若虫及不完整标本更是难以鉴别的难题,本文研究了它的分子标记快速检测方法。利用螺旋粉虱的CO I基因序列设计1对特异性引物,其扩增片段为399 bp。结果表明:此引物只对螺旋粉虱CO I基因具有特异性扩增,而对其他粉虱类昆虫不具有特异性扩增条带;该引物不仅能快速准确检测成虫,而且对其他虫态(卵、若虫和拟蛹等)亦具有同样的检测能力;同时,此检测方法灵敏度高,对成虫的检测限达到1/4000头。该检测方法的建立,为有效防控螺旋粉虱在我国的进一步扩散蔓延和危害提供了技术保障。

The spiraling whitefly,Aleurodicus disperses Russell,is a worldwide invasive pest.Morphological identification of adult of this pest is limited by its small size and the high degree of similarity with other whitefly.While morphological identification of egg,nymph,or half-baked specimen is almost impossible.In this study,a PCR method for rapid detection of A.disperses was established based on CO I gene sequence.A single fragment of 399 bp in length was amplified from all A.disperses individuals by using a pair of specific primers,which cannot be amplified from other closely related whitefly species.And it was proved to be valid for single spiraling whitefly egg,nymph 1st to 4th instar,male and female adults even the extraction of a single female adult of A.disperses was diluted to 1 /4000.The establishment of the method for rapid detection of A.disperses provided a technical support in prevention its spread and in reduction its harmfulness in China.