WANG Yingzhe,WANG Yinan,REN Wei,et al.Transformation of Alfalfa With Bar+CP4EPSPS Genes and Resistance Indentification[J].Northern Horticulture,2019,43(18):32-39.[doi:10.11937/bfyy.20184504]
Bar+CP4EPSPS基因转化紫花苜蓿的研究及抗性鉴定
- Title:
- Transformation of Alfalfa With Bar+CP4EPSPS Genes and Resistance Indentification
- 关键词:
- 紫花苜蓿; 转化; 农杆菌; Bar基因; CP4EPSPS基因
- Keywords:
- alfalfa; transformation; Agrobacterium; Bar gene; CP4EPSPS gene
- 文献标志码:
- A
- 摘要:
- 为了提高苜蓿抗除草剂的能力，以“公农1号”紫花苜蓿为试验材料，采用农杆菌介导的方法将Bar基因和CP4EPSPS基因转入紫花苜蓿中，研究了优化影响转化的5个主要因子（分别为外植体、农杆菌种类、侵染浓度、共培养时间、筛选浓度），建立了农杆菌介导的“公农1号”紫花苜蓿高效转化体系，获得转基因植株。结果表明：选择再生苗叶片作为外植体、EHA105农杆菌侵染、菌液浓度OD600值为0.8、共培养3 d、转化过程中草铵膦筛选浓度选择2 mg?L-1为宜，是最佳转化方案。试验结合分子检测和试纸条检测初步证明目的基因已经整合到紫花苜蓿中，最终喷施草甘膦确定12株为转基因植株。
- Abstract:
- In order to improve the herbicide tolerance of alfalfa,Bar gene and CP4EPSPS gene was transfected into ‘Gongnong No.1’ alfalfa using Agrobacterium-mediated method.The five main factors(such as explants，Agrobacterium species，infection concentration，co-culture time，screening concentration) affecting transformation efficiency were studied and established the ideal transformation system.The results showed that the optimal transformation scheme was to select the regenerated seedling leaves as explants，EHA105，bacteria concentration was OD600=0.8，co-cultivation time was 3 days，the concentration of glufosinate was 2 mg?L-1.Target genes were preliminarily proved to integrate into the genome of alfalfa by molecular identification and strip test.Finally，12 transgenic plants were identified by spraying glyphosate.
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备注/Memo
第一作者简介：王英哲（1985-），女，博士，助理研究员，研究方向为牧草转基因育种。E-mail:yingzhe120@163.com.责任作者：郝东云（1962-），男，博士，研究员，研究方向为植物分子生物学。E-mail：dyhao@cjaas.com.基金项目：吉林省农业科学院创新工程（人才基金）资助项目（c7208000102）；中国农业科技东北创新中心博士后基金资助项目；吉林省农业科学院结余经费（统筹部分）资助项目（y81990501）；现代农业产业技术体系建设专项资金资助项目（CARS-35-02）；吉林省科技厅重点科技研发资助项目（20180201072NY）。收稿日期：2019-03-12