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紫花苜蓿组织培养体系的建立及其遗传转化

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Keywords: 紫花苜蓿,组织培养,高频再生,遗传转化,农杆菌,卡那霉素,OD600值

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Abstract:

为建立高效的农杆菌介导的紫花苜蓿的遗传转化体系,以紫花苜蓿无菌苗叶片作为外植体,用8种含有不同激素浓度梯度的培养基对其进行愈伤组织及不定芽的诱导,结果表明,M7培养基(MS+1mg/L2,4-D+2mg/LCH+0.25mg/LKT+2mg/L琼脂)的愈伤组织诱导率最高,可达100%;继代表现最好,为胚性愈伤组织;同时,继续培养后可直接分化出不定芽,分化率达100%。结果表明,最适宜条件分别为卡那霉素(Kan)筛选浓度为75mg/L;农杆菌菌液OD600值为0.45-0.6之间。

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