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农杆菌介导的蓖麻转化体系优化

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.05.022, PP. 140-145

Keywords: 蓖麻,农杆菌侵染,转化体系

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Abstract:

针对影响农杆菌侵染效率的主要因素进行条件优化,确立了以OD600为0.8的菌液侵染、预培养5d后的外植体为最佳侵染条件;侵染外植体经过5d的共培养并除菌后,转到含有250mg/LKan的培养基中进行再生诱导,获得了多株具有Kan抗性的再生植株;通过在共培培养基中添加20mg/LAS,124mg/LNa2S2O3和152.4mg/LDTT,有效地抑制了侵染后外植体的褐化现象,提高了筛选阶段抗性芽的成活率;对具有Kan抗性的再生植株进行PCR和Southernblot验证,共获得18株包含外源基因的阳性转化株。

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