根癌农杆菌介导sHSP基因对半夏的遗传转化

Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica(2012)

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Abstract
目的:建立高效的半夏遗传转化体系。方法:以半夏试管苗的叶柄为外植体,采用根癌农杆菌介导法,探索了酚类物质、农杆菌菌液浓度、侵染时间、预培养时间及共培养时间等对半夏遗传转化效率的影响。结果与结论:不经过预培养阶段,用含AS 40 mg.L-1的根癌农杆菌菌液侵染15 min后共培养3 d时可有效提高遗传转化效率。将转化后的叶柄接到含有100 mg.L-1Kan和350 mg.L-1Carb的分化培养基上进行筛选培养,30 d左右在叶柄的两端分化出抗性的试管小块茎。对抗性转化植株进行Gus染色和PCR检测,结果表明外源基因sHSP已经整合到半夏基因组中。
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Key words
agrobacterium tumefaciens mediation,pinellia ternate,shsp,genetic transformation
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