核盘菌液滴凝集素SSL-6基因过表达质粒构建与油菜转化  

Over-expression Plasmid Construction of Lectin Gene SSL-6 from Sclerotinia sclerotiorum's Droplet and Rapeseed Transformation

作　　者：李浪[1] 刘正立[1] 刘询[2] 刘春林[1] Li Lang Liu Zhengli Liu Xun Liu Chunlin(Agricultural College, Hunan Agricultural Univesity, Changsha, 410128 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128)

机构地区：[1]湖南农业大学农学院,长沙410128 [2]湖南农业大学生命科学与技术学院,长沙410128

出　　处：《分子植物育种》2017年第1期160-165,共6页Molecular Plant Breeding

基　　金：国家支撑计划项目(2013BAD01B03-10)资助

摘　　要：菌核病是由腐生型真菌核盘菌寄生引起的一种世界范围性严重病害,是我国油菜的主要病害之一,严重影响油菜的品质和产量。目前没有找到针对核盘菌的有效的免疫资源,因此通过不同的途径来开展抗菌核病抗性资源筛选研究具有非常重要的现实意义。本研究本着"以毒攻毒"的思路,将核盘菌自身可能参与致病的分泌型凝集素蛋白SSL-6基因转入甘蓝型油菜中,以期获得具有抗菌核病的油菜种质资源。通过基因克隆、过表达载体构建和借助根癌农杆菌介导的油菜遗传转化,将SSL-6基因转到甘蓝型油菜"中双6号"基因组中;经过潮霉素抗性筛选与PCR鉴定,成功获得了75株转基因油菜;进一步RT-PCR检测,筛选到5株SSL-6基因高表达的转基因植株。这些转基因油菜植株为下一步菌核病的抗性研究提供了重要的基础材料。Sclerotinia stem rot caused by the parasitic saprophytic fungal-Sclerotinia sclerotiorum（Lib.） de is a worldwide serious disease and one of the major diseases that seriously affect the yield and quality of rapeseed in China. At present, the complete resistant resource to sclerotinia has not been found. Therefore, it has a realistic significance to carry out the research on screening of sclerotinia-resistant gene resource by different ways. In this study, based on the idea ＂to counteract one toxin with another＂, the secretive lectin-like protein SSL-6 gene from Sclerotinia sclerotiorum was transformed into rapeseed, in order to obtain the sclerotinia-resistant rapeseed germplasm. By gene cloning, over-expression vector construction and genetic transformation via Agrobacterium tumefaciens, SSL-6 gene was transferred into Brassica napus ＂No.6 Zhongshuang＂genome. Through resistance selection of hygromycin and PCR identification of hygromycin-resistant plants, seventy-five transgenic plants were successfully gained. Among the transgenic plants, 5 plants with SSL-6 gene over-expression were identified by RT-PCR, which provide basically materials for further research of resisting to Sclerotinia.

关 键 词：核盘菌 分泌液滴 凝集素基因 过表达质粒 油菜转化 

分 类 号：S435.654[农业科学—农业昆虫与害虫防治] Q943.2[农业科学—植物保护]