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棉花学报  2013 

基于抗草甘膦基因的棉花茎尖农杆菌介导转化方法的研究

DOI: 1002-7807(2013)05-0410-07, PP. 410-416

Keywords: 农杆菌,棉花,茎尖,转化,草甘膦

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Abstract:

以中棉所49、珂字棉201和YZ-1为受体材料,通过对培养基草甘膦浓度、农杆菌侵染时间和浓度、共培养时间以及恢复培养条件等因素的优化,建立了基于抗草甘膦基因的棉花茎尖农杆菌转化技术体系,并将抗草甘膦基因(EPSPS-G6)导入3个受体材料,获得转基因抗草甘膦棉花植株。研究结果表明,适合于棉花茎尖农杆菌介导的草甘膦筛选浓度为10mg·L-1;茎尖转化体系为农杆菌菌液OD600为0.9~1.0,侵染时间为20min,共培养时间为48h,选用SH培养基并加入适量活性炭(0.5g·L-1)作为恢复培养基。用本研究创制的转化技术体系,转化处理3个陆地棉受体360个茎尖,共获得60株抗性再生植株,经PCR检测,获得阳性抗性植株26株,移栽成活23株。以茎尖外植体数计算,本体系的转化成功率6.4%。该转化体系适合于转化抗草甘膦基因或以抗草甘膦基因为筛选基因的外源基因,具有转化频率高、嵌合体少、转化周期短等优点。

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