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大豆科学  2015 

农杆菌介导大豆子叶节遗传转化体系的优化

DOI: 10.11861/j.issn.1000-9841.2015.05.0768, PP. 768-775

Keywords: 大豆,子叶节,农杆菌中文,转基因体系优化

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Abstract:

利用农杆菌菌株EHA105,对17个栽培大豆品种的大豆子叶节进行了侵染,从大豆基因型、氯气灭菌时间、外植体状态、菌株活力、侵染浓度、侵染时间、共培养时间等方面进行了优化。结果表明:氯气灭菌14~18h,可在不影响大豆种子活力的同时最大限度的减少污染;暗处理1d的外植体活力高于光照处理5~7d的外植体;菌液浓度(OD600nm)在0.8~1.0且侵染浓度(OD600nm)为0.6~0.8时GUS瞬时表达率最高;适宜的侵染时间和共培养天数分别为30min和4d。在上述优化研究基础上,形成一套综合的转基因体系,该体系的最高转化率可达3.33%。不同处理下的GUS瞬时表达率以及17个大豆品种的丛生芽诱导率综合评价显示,适宜转化的基因型为TL-1、HC-3、HC-6、Williams82。

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