花生iso-Ara h3基因在种子抗黄曲霉中的功能研究

项目名称： 花生iso-Ara h3基因在种子抗黄曲霉中的功能研究

项目编号： No.31200211

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 微生物学、植物学

项目作者： 王通

作者单位： 山东省花生研究所

项目金额： 22万元

中文摘要： 花生的黄曲霉毒素污染是限制花生产业发展和危害人类食品安全的重要因素。由于花生抗黄曲霉分子遗传机制不明，目前国内外花生抗性育种一直进展缓慢。培育耐旱和抗黄曲霉品种是最有效、经济的解决措施。本项目拟以前期花生蛋白质组学研究为基础，克隆与干旱胁迫、黄曲霉侵染相关的花生种子基因iso Ara h3。通过原核表达鉴定iso Ara h3基因的酶活性及体外抑菌功能。通过构建植物超表达载体和反义表达载体，转化花生和拟南芥，对转基因植株进行相关的基因表达和抗性分析，明确iso Ara h3在花生抗黄曲霉和耐旱中的功能。通过对iso Ara h3基因的上游顺式作用因子进行系统研究，明确其调控途径，并在此基础上结合生物信息学方法建立以重要基因为主线的花生抗黄曲霉调控网络加权矩阵模型。本项目的实施对于阐述花生抗黄曲霉调控途径、解决花生黄曲霉污染和

中文关键词： 花生；黄曲霉菌；基因表达；纯化；启动子

英文摘要： Aflatoxiin contamination of peanut (Arachis hypogaea. L) seeds is one of the most severe food safety factors that impair peanut productivity and human and animal health, especially in arid and semi-arid tropical areas. The development of host-plant resistance would be an effective approach to eliminate aflatoxin accumulation in peanut. Unfortunately, the progress in transferring the resistance genes from these resistant lines into commercial cultivars has been slow, due to lack of understanding of the resistance mechanism and markers associated with resistance. In this study, a peanut seed iso-Ara h3 gene, which had been identified to associate with drought stress and Aspergillus infection in our previous study, was cloned. And than iso-Ara h3 was inserted into pET28a vecter to obtain recombinant protein and study the enzyme activity and in vitro function in resistance to Aspergillus flavus infection. To identify the in vivo-function of iso-Ara h3 in peanut and Arabidopsis in responding to drought and Aspergillus flavus stresses, this gene was cloned into pCAMBIA1301 vecter and than transformed into peanut or Arabidopsis. And some resistance associated genes in the transgenic plants were detected too. Moreover, a comprehensive analysis of promoter of iso-Ara h3 was performed to display signal regulation in respo

英文关键词： peanut；Aspergillus flavus；gene expression；purification；promoter