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RNAi已经成为揭示基因功能的一个常用的有力工具。在昆虫RNAi技术体系中,通过显微注射方法向昆虫体内注入dsRNA,实现系统性干扰,是最为广泛采用的方法。但对于体型较小的昆虫,注射法操作存在较大的难度,特别是非常微小的昆虫烟粉虱Bemisia tabaci。为了进行烟粉虱大规模基因功能验证,需要开发一套成熟的RNAi技术体系。本研究以烟粉虱的CYP6CM1基因为目标基因,通过显微注射法注射不同浓度的dsRNA到烟粉虱4龄若虫,荧光定量PCR检测其沉默效果。结果表明:在对单头烟粉虱注射50 n L时,注射不同浓度的dsRNA,对CYP6CM1基因的沉默效果不同,最大沉默效率可达到80%以上。因此,本实验成功构建了烟粉虱的RNAi技术体系,也可用于其它粉虱类害虫的RNAi技术。
RNAi has become a commonly used and powerful tool for revealing gene function. In the insect RNAi technology system, it is the most widely used method to inject dsRNA into insects by microinjection to achieve systemic interference. But for the smaller size insects, the injection method has great difficulty, especially the very small Bemisia tabaci. In order to validate the large-scale gene function of B. tabaci, it is necessary to develop a set of mature RNAi technology system. In this study, the CYP6CM1 gene of Bemisia tabaci was used as the target gene, and different concentrations of dsRNA were injected into the 4th instar nymphs of B. tabaci by microinjection. The silencing effect was detected by quantitative PCR. The results showed that different concentrations of dsRNA were injected into a single strain of Bemisia tabaci (Gennadius) during 50 n L injection of different concentrations of CYP6CM1. The maximum silencing efficiency could reach more than 80%. Therefore, we successfully constructed the RNAi technology system of B. tabaci and used it for RNAi of other whitefly pests.