应用农杆菌介导法获得转大麦黄矮病毒GPV株系复制酶基因ORF1的小麦植株

Transformation of wheat with Barley yellow dwarf virus GPV replicase gene ORF1 via Agrobacterium-mediated method

以生产用4种基因型小麦为材料,利用农杆菌介导法将大麦黄矮病毒GPV株系复制酶基因ORF1分别转化愈龄40d的幼胚愈伤组织,经过G418筛选后,共再生出15株抗性植株,拓宽了小麦农杆菌转化的受体基因型。PCR及Southern杂交分析证实大麦黄矮病毒复制酶基因ORF1已经整合到小麦基因组中,共获得14株转基因植株。抗病性初步检测结果显示,转基因植株对GPV株系具有抗性。研究结果还表明,对于不同的基因型小麦进行转化时,需选择适合本基因型的菌株。

Barley yellow dwarf virus （BYDV） GPV replicase gene ORF1 was transferred to 40 days embryogenic calli from 4 genotypes of cultured wheat by Agrobacterium-mediated method. After the calli were cocultivated with Agrobacterium cells and selected with G418, 15 resistant plants were regenerated. The results of PCR and Southern blotting analysis demonstrated that the exogenous gene ORF1 had been integrated into the genome of 14 transgenic plants. The primary resistance tests showed that the transgenic plants were possessed of resistance tO GPV. The research also showed that it was better to select a suitable bacterium strain for different genotypes.