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作 者 ：唐俊,陈敏,刘万宏,张磊,廖志华

期 刊 ：中草药 2008年

关键词：半夏；根癌农杆菌；遗传转化；草甘膦；

Keywords:Pinellia ternata (Thunb.)Berit., Agrobacterium tumejaciens, genetic transformation glyphosate,

摘 要 ：目的将抗草甘膦突变基因(aroA-M12)转入半夏.获得草甘膦抗性植株。方法用携带植物表达载体pASM12的根癌农杆菌LBA4404对半夏进行叶盘法转化;在选择培养基中加入10 mg/L草甘膦进行筛选.获得草甘膦抗性的再生植株;对再生植株进行PCR检测和草甘膦抗性实验。结果PCR检测表明幼叶的转化频率达23%,叶柄的转化频率达33%;对PCR阳性植株进行草甘膦抗性实验,多数植株在不同程度上表现出对草甘膦的抗性。结论本实验建立了半夏的遗传转化方法及草甘膦筛选的转化体系。为培育抗除草剂的半夏新品种及草甘膦筛选体系在半夏基因工程中的应用奠定了基础。

Abstract:Objective To transfer the anti-glyphosate gene aroA-M12 into Pinellia ternate and get transgenic plantlets with resistance to glyphosate.Methods P.ternate was transformed via Agrobacteri- urn turnefaciens strain LBA4404 harboring the plasmid pASM12;Transgenic explants were directly select- ed on medium containing glyphosate 10 mg/L and transgenic plantlets were confirmed by genomic PCR techniques and glyphosate resistance test.Results PCR Analysis showed the transformation frequency of young leaf blades was 23% and petioles was 33%;Glyphosate resistance test indicated that most trans- genic plantlets could resist to glyphosate at various levels.Conclusion The establishment of the transfor- mation method facilitates P.ternate of breeding anti-herbicide varieties of P.ternate and applying the glyphosate-based selection system for P.ternate genetic engineering.

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