植物介导大豆食心虫RNA干扰的机制研究

项目名称： 植物介导大豆食心虫RNA干扰的机制研究

项目编号： No.31201229

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 食品科学、农学基础与作物学

项目作者： 孟凡立

作者单位： 东北农业大学

项目金额： 25万元

中文摘要： 大豆食心虫是危害东北大豆生产最严重的害虫，培育抗虫大豆品种是防治食心虫最经济有效的方法。但是，传统育种周期长，效率低。通过前期研究，我们首次利用RNAi技术将食心虫18SrDNA和P0dsRNA片段分别导入转入大豆品种，创制了高度抗食心虫的转基因大豆株系。然而，目前尚不清楚dsRNA介导食心虫靶基因沉默，提高大豆抗食心虫的机制。本项目在前期工作基础上，首先利用转录组测序，分析取食转基因和非转基因豆荚食心虫幼虫的基因表达谱变化，确定差异表达基因；进而：(1)通过酵母双杂交技术筛选与18SrDNA或P0互作的蛋白；(2)克隆食心虫SID和SR基因，通过RNA干扰技术，研究SID和SR基因在食心虫吸收转基因大豆表达的dsRNA途径中的功能。通过系统研究，阐明植物介导食心虫RNA干扰的机制，为培育新一代抗虫转基因大豆品种提供科学依据和实践基础。

中文关键词： 大豆食心虫；系统RNA干扰；转基因大豆；抗虫；

英文摘要： oybean pod borer (Leguminivora glycinivorella (Mats.) obraztsov) is one of the most serious soybean pests in Northeastern China. The hatched larvae enters the pod and uses the young beans as a food source until the formation of mature larvae. The larvae of soybean pod borer feed actively on the immature beans can cause a considerable yield and seed quality loss. The control of this pest is difficult due to the inability to achieve the contact between the pesticide and the larvae inside the soybean pods. Therefore, the breeding of pest-resistant variety of soybean is the most avaluable methodology to restrict the attack of soybean pod borer. However, the development of resistant variety is time cost and the effect of resistance is limited. Our previous results showed that the transformation of 18rDNA and P0dsRNA of soybean pod borer into soybean cultivar distinctly enhanced the plant resistance to pod borer. But how the fragments of 18rDNA and P0 dsRNA worked on against soybean pod borer remained unknown. The objectives of the present proposal are 1) to perform the transcriptom sequencing of the larvae RNA collected from transgenic soybean pods (integreded with the insect dsRNA) and non-transgenic pods, and to find the differential expressed genes. 2) to determine the protein interactions between 18rDNA/P0 protei

英文关键词： Soybean pod borer；systemic RNAi；Transgenic soybean；insect resistance；