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ORF2小麦植株

Transformation of wheat with Barley yellow dwarf virus GPV replicase gene ORF2 via Agrobacterium-mediated method

以生产用5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ORF2转化愈龄40d的幼胚愈伤组织,经过G418筛选后,共再生出9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ORF2已经整合到小麦基因组中。初步抗病性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,受体基因型、外植体的生理状态对农杆菌侵染起着至关重要的作用。

Barley yellow dwarf virus （BYDV） GPV replicase gene ORF2 was transferred to embryogenic calli of 40 days from 5 genotypes of cultured wheat by Agrobacteriurn-mediated method. After the calli was co-cultivated with Agrobacteriurn and selected with G418, 9 resistant plants were regenerated from the calli. The results of PCR and Southern analysis demonstrated that the exogenous gene ORF2 has been integrated into the genome of transgenic plants. The primary resistant examination showed that the transgenic plants have resistance to GPV isolate. The research showed that receptor genotype and the status of explants were important in wheat transformation.