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【背景】双钩巢粉虱是近年在我国南方新发现的一种入侵害虫,其寄主范围广,危害严重,进一步扩散蔓延趋势明显。【方法】针对粉虱类害虫体型微小、形态相似、难以准确快速识别的问题,以双钩巢粉虱为靶标,以我国常见的其他8种/隐种粉虱为参照,采用基于mtDNA COⅠ基因的种特异性SS-COⅠ方法,研究其快速分子检测技术。利用COⅠ通用型引物LCO-1490/HCO-2198获得双钩巢粉虱及其他常见粉虱的COⅠ序列,根据测序结果设计特异性SS-COⅠ引物1对(PPZYF1/PPZYR1),其扩增片段大小为233 bp。【结果】种特异性检验结果显示,该引物只对双钩巢粉虱的COⅠ基因具有扩增能力,对其他种/隐种粉虱,包括烟粉虱的不同隐种(MEAM1隐种、MED隐种、AsiaⅡ3隐种和AsiaⅡ1隐种)以及温室粉虱、柑橘粉虱、黑刺粉虱、螺旋粉虱等不具有扩增效果。该引物不仅对不同性别的成虫具有良好的扩增能力,对1~4龄若虫和单粒卵亦具有同样的扩增效能,其最低检测阈值为1/40960头雌性成虫。【结论与意义】本检测技术对口岸检疫以及观赏植物、果树及其种苗调运中的害虫检测、监测意义重大。
【Background】 The double-hooked whitefly, whitefly, is a newly discovered invasive pest in south China in recent years. Its host range is wide and its harm is serious. The trend of spreading and spreading is obvious. 【Method】 In view of the small size and similar morphological characteristics of whitefly pests, it is difficult to accurately and rapidly identify the whitefly pests. The target genes of B. tabaci were compared with the other 8 species of C. occidentalis in our country. Based on the mtDNA COⅠ gene Species-specific SS-COⅠ method to study its rapid molecular detection technology. COⅠ sequences of B. tabaci and other common whiteflies were obtained by COⅠ universal primer LCO-1490 / HCO-2198. One pair of primers (PPZYF1 / PPZYR1) was designed according to the sequencing results. The amplified fragment size was 233 bp. 【Result】 The result of species-specific test showed that this primer could amplify only the COⅠ gene of B. tabaci, but had no effect on the other species (Cryptocaryon Bemisiae) AsiaⅡ3 hidden species and AsiaⅡ1 hidden species), as well as green house whitefly, citrus whitefly, black tick, whitefly, etc. do not have the amplification effect. The primer not only has good amplification ability for adults of different sexes, but also has the same amplification efficacy on the first to the fourth instar nymphs and single eggs, and the minimum detection threshold is 1/40960 female adults. [Conclusions and significance] The detection technology for port quarantine and ornamental plants, fruit trees and their seedlings in the pest detection, monitoring of great significance.