野生番茄LA2157中热稳定抗根结线虫基因Mi-9候选基因的分离与过表达载体构建  

Isolation of Heat-stable Candidate Gene Mi-9 for Resistance to Root-knot Nematode in Wild Tomato LA2157 and Construction of Over-expressive Vector

作　　者：王银磊[1,2] 陈伟男 赵丽萍 周蓉[1,2] 宋刘霞 余文贵[1,2] 赵统敏 WANG Yin-lei;CHEN Wei-nan;ZHAO Li-ping;ZHOU Rong;SONG Liu-xia;YU Wen-gui;ZHAO Tong-min(Institute of Vegetable Crops,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,Jiangsu,China;Key Laboratory for High Efficient Horticulture Crops Genetic Improvement of Jiangsu Province,Nanjing 210014,Jiangsu,China;School of Horticulture and Plant Protection,Yangzhou University,Yangzhou 225009,Jiangsu,China)

机构地区：[1]江苏省农业科学院蔬菜研究所,江苏南京210014 [2]江苏省高效园艺作物遗传改良重点实验室,江苏南京210014 [3]扬州大学园艺与植物保护学院,江苏扬州225009

出　　处：《中国蔬菜》2017年第12期30-34,共5页China Vegetables

基　　金：国家自然科学青年基金项目(31401884);江苏省自然科学青年基金项目(BK20140739)

摘　　要：根结线虫是番茄上的重要土传病害,选育抗根结线虫品种是最有效的防治方法,但是目前转育到栽培番茄中的抗根结线虫基因Mi-1在土温高于28℃时就丧失了抗性。本试验利用热稳定抗根结线虫野生番茄材料LA2157,根据Mi-1基因的序列信息,对其中热稳定抗根结线虫Mi-9基因进行同源克隆,在LA2157中共获得2个候选基因片段;通过In-Fusion克隆技术,将候选基因与过表达载体p BI121进行连接,经电泳检测和测序分析,最终构建Mi-9候选基因的过表达重组载体。Root-knot nematode disease is one of the important tomato soil-borne diseases.Breeding varieties with root-knot nematode resistance is an effective control method to deal with this disease.But at present,Mi-1the only gene transferred to tomato culture with resistance to root-knot nematode loses its resistance when the soil temperature is over28℃.In this study,wild tomato accession LA2157with heat-stable resistance to root-knot nematode was used.According to the sequence information of Mi-1,we carried out homologous cloning on Mi-9gene with heat-stable resistance to root-knot nematode and gained2candidate gene segments from LA2157.The candidate gene was ligated with the overexpression vector pBI121by In-Fusion cloning technique.Electrophoresis and sequencing analysis proved that the recombinant vectors of the candidate gene were constructed.

关 键 词：番茄 根结线虫 Mi-9基因 同源克隆 过表达载体构建 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]