基因枪法转Bt/GNA基因超甜玉米植株的获得及抗虫性鉴定

Transformation of Supersweet Corn with Bt/GNA Gene by Particle Bombardment and the Bioassay for Insect-resistance

以超甜玉米自交系S1幼胚诱导的愈伤组织为受体材料,利用基因枪法共转化Bt和GNA基因,获得了128株再生植株,移栽成活31株,其中26株可育。经PCR检测外源基因整合进玉米基因组中,含Bt基因11株,其中有5株能扩增出GNA基因。Southern杂交证明,目的基因已经整合在玉米基因组中。对9株转基因T1代植株叶片室内接虫抗性鉴定,结果显示,5株转基因植株叶片饲喂6d后明显抑制玉米螟幼虫的发育,幼虫体重增加较少,幼虫死亡率高,表现较强的抗性。

The calli, derived from immature embryos of the inbred line S1, were transformed with the plasmids DNA containing Bt/GNA gene via the particle bombardment-mediated method. 128 regenerated plants were obtained, 31 plants of which were survived through transplanting, 26 plants of which were fertile. PCR analyses of the To individuals demonstrated that, Bt gene was integrated into the genomes of 11 plants, GNA gene was into 5 plants. It was further concluded that Bt/GNA gene was integrated into the genomes of the regeneration plants by southern hybridization analysis. In addition, the insect resistance has been investigated in 9 T1 plants leaves by feeding the corn borers in a lab. The results showed that, the developments of larvae in 5 T1 transgenic plants were retarded compared with those larvae fed with non-transgenic control plants after 6 days, the average weight of larvae increased few, also showed that the rates of larvae mortality by feeding the T1 plants leaves were higher than those of non-transgenic control plants, and they remarkably resisted to the corn borers.