Bcutra在瓜实蝇性别决定和遗传工程防治中的作用研究

瓜实蝇（Bactrocera cucurbitae）是危害我国热带和亚热带地区果蔬类作物重要害虫之一，在海南省危害特别严重。我们已经鉴定瓜实蝇性别决定基因transformer（Bcutra）并发现了其性别特异的可选择性剪接方式。本项目拟利用CRISPR/Cas9技术敲除Bcutra，研究Bcutra突变对瓜实蝇生殖器官发育和生殖能力的影响，确定该基因在性别决定中的作用。然后利用该基因性别特异剪接方式并结合四环素表达控制系统和细胞毒性蛋白tTAV构建可控型雌性致死表达载体，通过转基因技术获得遗传工程修饰的瓜实蝇，筛选出雌性致死效率达100%但雄性生长发育正常的品系，研究该品系对瓜实蝇种群繁殖的影响。通过本项目研究，可望阐明Bcutra在瓜实蝇性别决定和遗传工程防治中的作用，为将来应用于瓜实蝇及其它害虫防控奠定理论和应用研究基础。

Bactrocera cucurbitae is one of very destructive pests in tropical and sub-tropical areas, particularly in Hainan province. We have previously identified the sex determination gene transformer (Bcutra) in Bactrocera cucurbitae and found its sex-specific alternative splicing mode. In the project Bcutra will be knocked-out using CRISPR/Cas9 technology, the influence of Bcutra on the development of reproductive organs and on the reproductive ability of Bactrocera cucurbitae will be studied after Bcutra is mutated, and then its role in sex determination of Bactrocera cucurbitae will be determined. We will obtain the controllable female-lethal construct by combining sex-specific splicing mode of Bcutra, tetracycline on/off system of controlling gene expression, and cellular toxic protein tTAV; and obtain genetically engineered Bactrocera cucurbitae lines with 100% female-lethality but developmentally normal males, and study the control effects of these lines to the pest population. The research in this project will clarify the functions of Bcutra in sex determination of Bactrocera cucurbitae and its genetic engineering control, therefore providing the theoretical and practical basis of genetic engineering control method for Bactrocera cucurbitae and other pests.