转基因技术介导的美国白蛾性别决定基因tra2的功能研究

Transformer2(tra2) 基因是昆虫性别决定和生长发育的重要参与因子，对性别决定、胚胎和生殖腺发育、翅二型分化等具有重要的生物学功能。除家蚕外，tra2基因在鳞翅目害虫中的功能所知甚少。申请人前期研究发现敲除美国白蛾tra2基因可导致蛹期生殖孔畸形，与家蚕表型不同，但其具体的分子机制尚不清楚。因此，本项目拟从以下几个方面进一步探讨白蛾tra2基因的生物学功能：（1）tra2基因的表达模式及其性别、时期和组织特异性的剪接体状况分析；（2）通过转基因技术构建tra2基因不同剪接体的敲除突变体，深入探究其在性别决定和生长发育中的功能分化；（3）通过RNA-seq深入挖掘tra2突变引起的差异基因表达和分子信号转导；利用co-IP和基因敲除进一步分析下游基因的分子生物学功能。研究将有助于深入理解tra2基因在美国白蛾性别决定和生长发育中的分子机制，并对开发新型害虫防治方法提供理依据。

Transformer2 (tra2) gene is an important factor involved in insect sex determination and growth and development, which has important biological functions for sex determination, embryonic development, gonadal development and wing dimorphism. The biological function of tra2 gene in other lepidopteran pests, is rarely studied, except for Bombyx mori. Different from the silkworm, the previous study have found that knockout the tra2 gene could lead to abnormalities of pupal gonopores, but its specific molecular mechanism is still unclear in Hyphantria cunea. Therefore, this project intends to further explore the biological functions of the tra2 gene of H. cunea from the following aspects: (1) Analyses of the expression pattern of tra2 gene and its specific splicing isoforms in different genders, periods, and tissues of H. cunea. (2) Construction of knock-out mutants of different tra2 splicing isoforms through transgenic technology to explore its specific functions in sex determination and growth and development. (3) Deeply exploring the differential gene expression and molecular signal transduction caused employing RNA-seq technology in the tra2 mutants. Then, to further verify the molecular biological functions of downstream genes using co-IP and knockout assay. Our work will help to deeply understand the molecular mechanism of tra2 gene in the sex determination, growth and development of H. cunea, and provide a theoretical basis for the development of new pest control methods.