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为了解参与烟粉虱免疫反应的clip丝氨酸蛋白酶(clip-domain serine proteases in Bemisia tabaci,Bt CLIPs)基因,对已鉴定的8个Bt CLIPs基因编码的蛋白进行序列分析,与冈比亚按蚊、家蚕、黑腹果蝇、烟草天蛾4种模式昆虫的同源蛋白进行系统进化分析,并通过q RT-PCR检测球孢白僵菌侵染烟粉虱4龄若虫后Bt CLIPs的时间表达模式。结果显示,8个Bt CLIPs基因均为含有CLIPs结构域的丝氨酸蛋白酶基因;Bt CLIP19、Bt CLIP20和Bt CLIP21独自聚在进化树的一支上;其它Bt CLIPs均与4种模式昆虫中的1种或多种同源蛋白聚在一起;与对照相比,8个Bt CLIPs基因受球孢白僵菌侵染后在不同时间的相对表达量均上调,但诱导程度不同,48 h的相对表达量达最高,其中Bt CLIP19上调表达最明显,为对照组的71倍。研究表明,8个Bt CLIPs可能参与烟粉虱对真菌侵染的免疫反应,并成为烟粉虱生物防治的新靶标。
In order to understand the biosynthesis of the Bt CLIPs genes involved in the immune response of Bemisia tabaci (GenBank), the sequences of the eight Bt CLIPs genes identified were sequenced and compared with those of Anopheles gambiae, Bombyx mori, Drosophila melanogaster and Homoptera exigua were analyzed phylogenetically. The temporal expression pattern of Bt CLIPs in Beauveria bassiana against 4th instar nymphs of Bemisia tabaci was detected by q RT-PCR. The results showed that the eight Bt CLIPs genes were both serine protease genes containing CLIPs domain. Bt CLIP19, Bt CLIP20 and Bt CLIP21 clustered alone in one of the phylogenetic trees. The other Bt CLIPs were all associated with one of the four model insects Compared with the control, 8 Bt CLIPs genes were up-regulated by Beauveria bassiana infection at different times, but the degree of induction was different, the relative expression level at 48 h Up to the highest, of which Bt CLIP19 up-regulated expression of the most obvious, 71 times the control group. Studies have shown that eight Bt CLIPs may be involved in the immune response of B. tabaci to fungal infection and become a new target of biocontrol of B. tabaci.