利用mtCOIPCR-RFLP技术鉴定中国境内九个烟粉虱隐种

Identification of nine cryptic species of Bemisia tabaci ( Hemiptera: Aleyrodidae) from China by using the mtCOI PCR-RFLP technique

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 containing over 31 cryptic species, some of which are worldwide invasive insect pests. At present, two invasive cryptic species and 13 indigenous cryptic species have been recorded from China. Rapid and efficient methods for identifying cryptic species are important for investigating the population abundance of development of management strategies. However, apart from sequence BLAST search, an efficient method for indentifying various cryptic species in the field and the method of sequencing mtCOI and gene multiple species is still lacking at present. In this study, we adopted the PCR-RFLP technique, using one or more of the five restriction enzymes, namely TaqI, VspI, Van91I, NcoI and FokI, to identify nine whitefly cryptic species from China. The results showed that two cryptic species MEAM1 and China 1 could be identified through digestion with only TaqI, two cryptic species MED and Asia 1 could be identified through sequential digestion with TaqI ＋ NcoI, two cryptic species Asia II 3 and Asia II 9 could be identified through sequential digestion with TaqI ＋ Van91I, and three cryptic species Asia II 1, Asia II 6 and Asia II 7 could be identified through sequential digestion with TaqI ＋ VspI ＋ FokI. This study provides an efficient method for identifying nine cryptic species of B. tabaci from China.