高赖氨酸蛋白基因导入小麦的研究

为了培育高赖氨酸含量的小麦新材料,利用基因枪转化法将辣椒高赖氨酸蛋白Cflr基因导入到小麦品种扬麦16中,轰击了2 500枚小麦幼胚,获得抗除草剂Basta的再生植株176株(遗传转化采用的筛选标记基因为bar基因),经PCR鉴定Cflr基因阳性的转基因植株为23株.T1代幼苗喷洒130 mg·L-1的Basra溶液进行除草剂抗性鉴定,发现T1代幼苗的除草剂抗性出现了分离.PCR-Southern杂交进一步证明外源高赖氨酸蛋白基因Cflr基因已经整合到小麦的基因组中.通过种子赖氨酸含量和总蛋白质含量的测定,发现75.00％的转基因株系的赖氨酸含量高于受体扬麦16,77.78％的转基因株系的蛋白质含量高于受体扬麦16.本试验获得了一批赖氨酸含量提高的小麦转基因株系,这些高赖氨酸含量的转基因株系可进一步用于小麦高赖氨酸分子育种研究.

Obtaining and Detection of Transgenic Wheat Plants with High Lysine Content

In order to obtain transgenic wheat with improved lysine content in seeds, Cflr gene from pepper was introduced into wheat variety Yangmai 16 with particle bombardment.2 500 young embryos were bombarded and 176 Basta resistant regenerated plantlets were obtained.The PCR detection showed that the Cflr gene had been integrated into 23 wheat plantlets of Yangmai 16.Through spraying 130 mg·L^-1 Basta herbicide on the plantlets of the T₁ progenies,the Basta resistance of the T₁ progenies was tested and diverged among the T₁ generation plants.PCR Southern identification also confirmed that the Cflr gene had been integrated into wheat genome.The analysis of lysine content and protein content in seeds of the transgenic lines showed the lysine content in 75.00% transgenic lines and the protein content in 77.78% transgenic lines were higher than that in Yangmai 16 respectively.These transgenic lines could be further used in molecular breeding of wheat with high lysine content.