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烟粉虱Bemisia tabaci(Gennadius)是一个物种复合体,包括31个以上形态上无法区分的隐种,其中少数隐种是世界性入侵害虫。目前,在中国境内分布有2个入侵隐种和13个土著隐种。快速、高效的鉴别方法对掌握烟粉虱田间发生规律及制定相关防控策略具有重要意义。然而到目前为止,除了mtCOI基因测序比对外,尚未有一种简便的方法可以有效地区分多个烟粉虱隐种。本研究采用mtCOI PCR-RFLP技术,单独或组合使用TaqI,VspI,Van91I,NcoI和FokI这5种限制性内切酶,酶切烟粉虱mtCOI的PCR扩增片段,鉴别分布在中国境内的9个烟粉虱隐种。结果表明,单独使用TaqI酶切,可鉴别出MEAM1和China1两个隐种,使用TaqI+NcoI分步酶切可鉴定出MED和Asia1两个隐种,使用TaqI+Van91I分步酶切可鉴定出Asia II3和Asia II9两个隐种,使用TaqI+VspI+FokI分步酶切可鉴定出Asia II1,Asia II6和Asia II73个隐种。本研究为高效鉴别中国境内的多个烟粉虱隐种提供了方法。
Bemisia tabaci (Gennadius) is a species complex that includes more than 31 morphologically indistinguishable species, of which a few species are worldwide invasive pests. At present, there are 2 invaders and 13 indigenous hidden species distributed in China. The rapid and efficient identification method is of great importance to understand the field occurrence and control strategy of Bemisia tabaci. However, so far, except for the mtCOI gene sequence alignment, there is no easy way to effectively distinguish multiple whitefly bug occultities. In this study, mtCOI PCR-RFLP technology was used to digest the mtCOI fragment of Bemisia tabaci by using five restriction enzymes TaqI, VspI, Van91I, NcoI and FokI separately or in combination. Bemisia tabaci hidden species. The results showed that two recessive species of MEAM1 and China1 could be identified by TaqI digestion alone. The two hidden species of MED and Asia1 could be identified by TaqI + NcoI digestion, and TaqI + Van91I digestion could be used to identify Asia II3 and Asia II9 two hidden species, using TaqI + VspI + FokI step digestion can be identified Asia II1, Asia II6 and Asia II 73 hidden species. This study provides a method for the efficient identification of multiple whitefly bugs in China.