花粉管通道法介导的抗大麦黄矮病毒转基因小麦研究

Transgenic wheat resistant to Barley Yellow Dwarf Virus obtained via pollen tube pathway

构建了大麦黄矮病毒GPV株系串联结构复制酶基因植物表达载体pPPI29,通过花粉管通道法转化了CA8686小麦,对T0代种子进行了分子检测,获得6株转基因小麦,转化率达到0.45%;对获得的T0代转基因小麦进行了田间抗病性鉴定,结果表明4株转基因小麦对GPV株系表现为高抗,可以推迟发病23 d。

The cDNA of the direct repeat replicase genes of BYDV-GPV was obtained by RT-PCR and was inserted into the plasmid to construct an expression plasmid named pPPI29. CA8686 was then transformed with pPPI29 via pollen tube pathway. Six positive transgenic wheat plants were confirmed by Dot-plot, PCR and PCR-Southem assay among 1343 plants and the transformation frequency was 0.45 %. The resistance evaluation of To generation of the transgenic wheat plants carrying GPV replicase gene were carried out. Compared with non-transgenic plants, four transgenic lines of CA8686 showed higher level of resistance after virus infection. They could delay symptom development for about 23 days.