转G10eve基因棉花的获得及草甘膦抗性初探  

Generation and Glyphosate Resistance Analysis of G10eve Transgenic Cotton

作　　者：陆国清 王春玲[2] 郝宇琼 郭惠明 黄英金[1] 程红梅[2] Lu Guoqing;Wang Chunling;Hao Yuqiong;Guo Huiming;Huang Yingjin;Cheng Hongmei(College of Agronomy,Jiangxi Agricultural University,Key Laboratory of Crop Physiology,Ecology and Genetic Breeding,Ministry of Education,Nanchang 330045,China;Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]江西农业大学农学院,作物生理生态与遗传育种教育部重点实验室,南昌330045 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《棉花学报》2018年第1期21-28,共8页Cotton Science

基　　金：国家转基因重大专项(2014ZX0800912B)

摘　　要：【目的】为培育高抗草甘膦棉花新品种提供种质材料。【方法】对草甘膦抗性基因G10eve进行生物信息学分析,利用农杆菌介导的方式将G10eve导入到珂字棉312中。利用PCR(Polymerase chain reaction)、qRT-PCR(Quantitative real-time PCR)方法分析G10eve的表达情况。对受体及转基因植株进行草甘膦喷施实验,测定其莽草酸含量,分析其抗性水平。【结果】最终获得28个独立的转基因阳性系,转化率及幼苗分化率分别高达49.3%、40.6%。PCR及qRT-PCR检测证明外源G10eve基因已经整合到棉花基因组中,且在不同转基因株系间表达量差异显著。在子叶期,转基因植株可耐受8 mL·L^(-1)浓度的草甘膦,而非转基因对照在2 mL·L^(-1)浓度草甘膦处理下已出现药害。草甘膦处理前后,转基因植株叶片莽草酸含量未出现明显积累,进一步说明其具有草甘膦抗性。【结论】过量表达G10eve基因能够提高受体材料对草甘膦的抗性,获得了高抗草甘膦的棉花株系。[Objective]Novel germplasm of cotton was generated for development of cultivars with high resistance to glyphosate.[Method]The glyphosate resistance gene G10eve was preliminarily analyzed using bioinformatics methods and introduced into'Coker 312'via Agrobacterium-mediated transformation.The expression of G10eve was estimated by PCR(Polymerase chain reaction)and qRT-PCR(Quantitative real-time PCR).The content of shikimic acid and tolerance to glyphosate were determined in response to application of glyphosate to'Coker 312'and transgenic plants.[Result]A total of 28 independent positive transgenic lines were obtained.The transformation efficiency and the rate of seedling differentiation were up to 49.3%and 40.6%,respectively.PCR and qRT-PCR analysis showed that G10eve was integrated into the cotton genome,and the G10eve expression level differed significantly among the transgenic lines.At the cotyledon stage,the transgenic cotton showed resistance to 8 mL·L-1 glyphosate,whereas'Coker 312'showed phytotoxicity to the 2 mL·L-1 glyphosate treatment.No significant accumulation of shikimic acid was detected in transgenic plants before or after glyphosate treatment,which indicated the typical physical characteristic of glyphosate resistance.[Conclusion]Overexpression of G10eve in cotton improved resistance to glyphosate.Thus,novel germplasm of cotton resistant to glyphosate was generated.

关 键 词：棉花 G10eve 草甘膦 抗性 

分 类 号：S562.034[农业科学—作物学]