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棉花学报 ›› 2018, Vol. 30 ›› Issue (1): 21-28.doi: 10.11963/1002-7807.lgqchm.20171129

• 研究与进展 • 上一篇    下一篇

G10eve基因棉花的获得及草甘膦抗性初探

陆国清1, 2,王春玲2,郝宇琼2,郭惠明2,黄英金1*,程红梅2*   

  1. 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
  • 收稿日期:2017-02-16 出版日期:2018-01-15 发布日期:2018-01-15
  • 通讯作者: 程红梅,chenghongmei@caas.cn;黄英金,yjhuang_cn@126.com
  • 作者简介:陆国清(1983―),女,博士研究生luguoqing007@163.com
  • 基金资助:
    国家转基因重大专项(2014ZX0800912B)

Generation and Glyphosate Resistance Analysis of G10eve Transgenic Cotton

Lu Guoqing1, 2, Wang Chunling2, Hao Yuqiong2, Guo Huiming2, Huang Yingjin1*, Cheng Hongmei2*   

  1. 1. College of Agronomy, Jiangxi Agricultural University/ Key Laboratory of Crop Physiology, Ecology and Genetic Breeding, Ministry of Education, Nanchang 330045, China; 2. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2017-02-16 Online:2018-01-15 Published:2018-01-15

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。

关键词: G10eve; 草甘膦; 抗性

Abstract: [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.

Key words: cotton; G10eve; glyphosate; resistance

中图分类号: 
  • S562.034