杆状病毒AcMNPV中Ac109蛋白的抗虫功能分析

Analysis of the Anti-insect Function of Ac109 Protein in Baculovirus AcMNPV

为研究苜蓿银纹夜蛾核型多角体病毒(AcMNPV)中病毒增殖因子Ac109对鳞翅目害虫甜菜夜蛾幼虫的抗虫效应,采用Bac-to-Bac杆状病毒表达系统构建了重组病毒AcMNPV-Ac109-EGFP。免疫印迹和荧光显微镜观察结果证实,重组病毒AcMNPV-Ac109-EGFP侵染昆虫Sf9细胞后Ac109的表达量和细胞的荧光强度随着侵染时间延长而显著增加。抗虫试验表明,与AcMNPV处理组、PBS对照组相比,AcMNPV-Ac109-EGFP病毒处理组的幼虫体质量增长缓慢,有较高的致死率和低的蛹化率及羽化率;AcMNPV与AcMNPV-Ac109-EGFP病毒处理组的最终死亡率分别为58.33%和73.33%;蛹化率分别为41.667%和25.000%;羽化率分别为20.00%和11.67%,说明Ac109可提高病毒对甜菜夜蛾幼虫的抗虫性。以上结果构建了1株高抗虫活性的重组病毒,并为这个重组病毒潜在的应用价值提供了试验依据。

To study the insecticidal efficiency of virus multiplication factor Ac109 in baculovirus insecticide Autographa californica multiple nucleopolyhedrovirus（AcMNPV）on the host Spodoptera exigua Hubner larvae, the Bac-to-Bac baculovirus expression system was used to generate recombinant baculovirus AcMNPV-Ac109-EGFP. Western blot and fluorescence detection confirmed the recombinant baculovirus AcMNPV-Ac109-EGFP was constructed successfully, and the fluorescence intensity and the expression level of Ac109 were enhanced significantly with the infection process. Anti-insect activity analysis showed that compared with PBS treatment group, the larvae in the AcMNPV-Ac109-EGFP treatment group was growing slowly, and had higher mortality rates, lower pupation rates and lower eclosion rates. The mortality rates of larvae in AcMNPV and AcMNPV-Ac109-EGFP treatment groups were 58.33% and73.33%, respectively, the pupation rates were 41.667% and 25.000%, respectively, and the emergence rate was 20.00% and 11.67%,respectively, which suggested that the recombinant baculovirus AcMNPV-Ac109-EGFP had high sensitivity to the Spodoptera exigua Hubner larvae. In this study, a recombinant baculovirus AcMNPV harboring ac109 gene with high anti-insect activity was constructed and the its insecticidal potential value was provided.