油菜乙酰乳酸合成酶抑制剂类除草剂抗性突变体M9 的遗传和基因克隆.pdfVIP

中国农业科学 2012,45(20):4326-4334 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.20.022 油菜乙酰乳酸合成酶抑制剂类除草剂抗性突变体 M9的遗传和基因克隆 胡茂龙，浦惠明，高建芹，龙卫华，戚存扣，张洁夫，陈 松 （江苏省农业科学院经济作物研究所/ 国家油料作物改良中心南京分中心/农业部长江下游棉花与油菜重点实验室，南京 210014 ） 摘要：【目的】分离和鉴定自然突变的油菜乙酰乳酸合成酶抑制剂类除草剂抗性突变体M9抗性基因，阐明抗 性产生的分子基础。【方法】通过配置杂交组合研究其遗传规律，应用同源序列法克隆油菜ALS家族基因，采用杂 交转育和小孢子培养技术将抗性基因转育到陆奥-五十铃细胞质雄性不育（MICMS）恢复系中，进行PCR鉴定。【结 果】该突变体抗性由 1 个显性核基因控制。克隆获得 BnALS1—BnALS3，核酸序列比对发现，M9 抗性是由 BnALS1 的点突变使蛋白序列的第638位丝氨酸（AGT）残基被天冬酰胺酸（AAT）替代所致，将此抗性基因命名为BnALS1R。 抗性基因 BnALS1R 转育到MICMS恢复系中的PCR检测表明，所有抗除草剂的恢复系都携带有 BnALS1R。【结论】突 变体M9抗性由 1个显性核基因控制，BnALS1 第638位丝氨酸突变成天冬酰胺酸是抗性产生的分子基础。 关键词：除草剂；甘蓝型油菜；乙酰乳酸合成酶；点突变；BnALS1R Inheritance and Gene Cloning of an ALS Inhabiting Herbicide- Resistant Mutant Line M9 in Brassica napus HU Mao-long, PU Hui-ming, GAO Jian-qin, LONG Wei-hua, QI Cun-kou, ZHANG Jie-fu, CHEN Song (Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Sub-center, National Center of Oil Crops Improvement/Key Laboratory of Cotton and Rapeseed (Nanjing), Ministry of Agriculture, Nanjing 210014) Abstract: 【Objective 】The objective of this study is to identify and characterize the resistant gene of a mutant line M9，which was previously found in the rapeseed (brassica napus L.) by spontaneous mutation, conferring resistance to acetolactate synthase or acetohydroxyacid synthase (ALS or AHAS) inhibiting herbicides, and understand the molecular basis for resistance to ALS-inhibiting herbicides. 【Method 】Reciprocal crosses were made in order to study the inheritance of resistance. Three genes BnALS1-BnALS3 encoding ALS were isolated from the mutant and wild type by using the homology-based candi