摘要
根癌农杆菌菌株Ａｇｌ Ⅰ的Ｔｉ 质粒ｐＵＮＮ－２ 带有Ｕｂｉ１ 启动子驱动的ｎｐｔ Ⅱ基因。７ 种基因型小麦幼胚或胚性愈伤组织用于农杆菌介导的转化实验。经过不同浓度巴龙霉素的筛选，３ 种基因型小麦产生抗性愈伤组织并再生植株。再生植株经ＰＣＲ 和Ｓｏｕｔｈｅｒｎ 杂交鉴定为转基因植株，转化频率（ 再生转基因植株的小麦愈伤组织数／ 用于转化实验的愈伤组织数） 为３．７％ ～５ ．９％ 。小麦基因型及转化材料的起始生理状态是影响ＴＤＮＡ转移的重要因素。
A highly efficient transformation of wheat mediated by Agrobacterium tumefaciens and plant regeneration is described. The strain of A. tumfaciens p UNN-2 / Agl Ⅰ carrying npt Ⅱ gene was used to transfer wheat ( Triticum aestivum L.). Immature embryos and embryogenic calli from 7 genotypes of cultured wheat were used for transformation. After the embryos and calli were co cultivated with Agrobacterium and selected with paromomycin (an analogue of kanamycin), resistant embryogenic calli and resistant plants regenerated from the calli of 3 genotypes of wheat were produced. Transgenic plants were determined by PCR amplification of transgene fragments and confirmed by Southern hybridization with an transformation efficiency (number of transgenic plants regenerated from independent calli relative to the number of infected calli) of 3.7%～5.9%. The genotypes and the state of a suitable starting material were shown to be important in the competence for T DNA transfer.
关键词
小麦
根癌农杆菌
介导法
nptⅡ基因
转基因植株
Agrobacterium tumefaciens,wheat (Triticum aestivum L.),nptⅡ gene,transgenic plants
