大豆对大豆花叶病毒抗性基因RSC15的精细定位、克隆以及功能分析

Analysis of recombination plants of residual heterozygous lines in RN-9×7605 was used to narrow the mapping region of resistant gene RSC15. Based on bioinformatics and QRT-PCR, the candidate genes were selected for inductive expression analysis. Tissue expression analysis and sub-cellular localization of the gene RSC15 that could be key gene in resistance response to SC15 strain in RN-9 were utilized to investigate the tissue specific expression pattern and the functional position of the protein expression in the cell, respectively. The transgenic and virus-induced gene silencing (VIGS) technique were used to examine and confirm the function of the gene RSC15.This project will provide an opportunity to further understand the molecular mechanism resistance of resistance to SMV in soybean, and play an important role in improving soybean resistance by biotechnology method.

本研究拟通过对RN-9（抗）×7605（感）剩余杂合系的单株分析，精细定位抗病基因RSC15；利用生物信息学的方法和荧光定量PCR技术，在定位区段内搜索候选基因并进行诱导表达差异分析，获得可能参与RN-9抗SC15株系抗性反应的基因RSC15；对该基因进行组织表达分析及亚细胞定位，明确该基因在大豆不同器官中的表达特征及其编码的蛋白在细胞中发挥功能的具体部位；利用转基因和病毒诱导基因沉默（VIGS）技术，阐明及验证该基因的功能。通过本项目的研究，有助于明确大豆抗SMV的分子机制，同时对利用生物技术改良大豆的抗性也具有重要应用价值。

通过对杂合单株自交获得的次级F2群体和F2:3家系进行分析，将抗病基因RSC15定位于2个SSR标记BARCSOYSSR_06 _0825和BARCSOYSSR_ 06_0835之间，定位区间物理距离约为150 kb，预测有10个基因。利用qRT-PCR 技术，分析大豆花叶病毒（SMV）诱导后7个候选基因在不同时间点的表达差异以及不同组织特异性表达分析，推测基因Glyma.06g182600可能与SMV抗性相关。同时，对4个候选基因进行了克隆测序，结果显示基因Glyma.06g182600、Glyma.06g175100和Glyma.06g184400是潜在的抗性候选基因。