大豆胞囊线虫抗性候选基因Rhg1的克隆及其功能验证  

Molecular cloning and functional identification of resistant candidate gene Rhg1 to soybean cyst nematode

作　　者：高慕娟[1,2] 宋雯雯[1,2] 韩雪[1,2] 王继安[1,2] 

机构地区：[1]东北农业大学农学院,哈尔滨150030 [2]大豆生物学省部共建教育部重点实验室,哈尔滨150030

出　　处：《东北农业大学学报》2012年第4期27-31,共5页Journal of Northeast Agricultural University

基　　金：国家自然科学基金(30471092);国家科技部转基因生物新品种培育重大专项(2009ZX08009-089B)

摘　　要：大豆胞囊线虫(Soybean cyst nematode,SCN)是大豆生产上一种危害严重的世界性害虫,给大豆的产量和品质造成极大的损失。大豆抗性品种选育是其防治措施中最经济、有效的方法。文章拟利用RT-PCR方法克隆得到大豆胞囊线虫抗性候选基因Rhg1,通过构建植物过量表达载体pCAMBIA3301/Rhg1,并采用根癌农杆菌介导的大豆子叶节方法转化大豆东农50。PCR检测草丁膦抗性植株,表明目的基因已经整合到了大豆基因组中;实时荧光定量PCR结果也进一步证实,目的基因在转基因植株中有较高水平的表达丰度。在胞囊线虫的侵蚀下,转基因植株体内的超氧化物歧化酶含量显著高于野生型植株,而丙二醛含量低于野生型植株。研究证实了Rhg1为大豆胞囊线虫的主抗基因,同时为大豆胞囊线虫的分子抗性育种提供理论基础。Soybean cyst nematode（SCN） is a serious and destructive pest in soybean production worldwide and causes great loss on its yield and quality.It has been an economical and effective method to breed resistant soybean cultivars for decreasing or avoiding its damage.Clone the Rhg1 gene through the RT-PCR method and constructed the overexpression vector pCAMBIA3301/ Rhg1 that was transformed into soybean Dongnong50 via Agrobacterium-mediated transformation of cotyledonary node.Glufosinate-resistant plants were detected to be positive by PCR.Furthermore,the Rhg1 gene in transgenic plants were identified to be a high expression level through Real-time quantify PCR.The SOD content of transgenic plant was significantly higher than that of wild types,while the MDA content of transgenic plant was lower than that of wild types.The results confirmed that the Rhg1 gene was crucial resistant gene and provided the basis on molecular resistant breeding of soybean cyst nematode.

关 键 词：大豆胞囊线虫 Rhg1基因 过量表达 超氧化物歧化酶 丙二醛 

分 类 号：S565.1[农业科学—作物学] Q78[生物学—分子生物学]