转酸性蛋白酶pepB基因玉米的分子鉴定与农艺性状分析

通过分子生物学技术把酸性蛋白酶pepB基因转入受体玉米自交系基因组中,培育转酸性蛋白酶pepB基因玉米新品系。以耐盐基因badh作为筛选标记性基因,经过抗性筛选,对T1、T2、T3代转基因植株进行PCR检测,得到14个T1代转基因阳性植株,32个T2代转基因阳性植株和27个T3代转基因阳性植株。Southern杂交结果表明,外源酸性蛋白酶基因已经整合进玉米基因组中。RT-PCR结果表明,酸性蛋白酶基因在受体玉米中获得表达,获得外源酸性蛋白酶pepB基因遗传表达的T3代转基因玉米株系。通过对T2、T3代转基因玉米各品系农艺性状的分析结果表明,转基因玉米各品系在株高、茎粗、穗长等农艺性状上与受体亲本没有差异,但生育期均有不同程度的缩短。

Analysis on the Molecular Detection and Agronomic Characters of Genetic Maize with Transforming pepB Gene

The pepB gene of acid protease was transformed into the maize by molecular technique,selected and cultivated transgenic plant lines which contain pepB gene.In this research,transgenic plants were screened with salt-tolerance gene badh,and detected by PCR.Fourteen positive transgenic plants in T1 generation were got by PCR detecting,and 32 plants in T2,and 27 positive transgenic plants in T3 generation.The result of southern blot confirmed that pepB had been integrated into maize genome.RT-PCR further indicated that pepB gene had been expressed in maize recipient.Consequently,transgenic plant lines of T3 generation were obtained,and that pepB gene could inherit in it.The result of the analysis about the transgenic maize lines of T2,T3 generation indicated that compared with the recipient parent,transgenic maize lines' growing period short-ended and have no differences on agronomic characters of plant height,stem diameter,ear length,etc.