根癌农杆菌介导抗菌肽基因转化大豆的研究

摘要

为利用抗菌肽基因培育转基因大豆抗病材料,采用根癌农杆菌介导法将抗菌肽基因和绿色荧光蛋白融合基因转入大豆胚尖外植体.结果表明,60mg/L的卡那霉素可以对转化植株进行有效的筛选；在共培养时添加200μmol/L乙酰丁香酮有利于抗菌肽基因的转化；在生根培养基中不添加卡那霉素更有利于转化苗的成活.目的基因特异的PCR检测和绿色荧光蛋白表达分析,发现抗菌肽基因已转入大豆,并得到表达.%A anti-microbial peptide and green fluorescent protein (GFP) fusion gene was transformed into soybean embryos tips via Agrobacterium-mediation. Result showed 60mg/L kanamycin could effectively select the transformed plants, and 200μmol/L acetosyringone in coculture medium could improve gene transduction efficiency. No kanamycin was added in rooting medium for it was found harmful to root. PCR result and expression of green fluorescent protein ( GFP) indicated that the target gene was transformed into soybean and correctly expressed.