The Cloning Research Of Rice Resistance Blast Gene From Chaling Wild Rice

Using the method of combination of traditional and modern biotechnology to protect, research and utilize the Chaling wild rice as hunan local varieties of wild rice is imperative.In this study,we took the total DNA from Chaling wild rice with pollen tube channel injection method into the receptor cultivated rice (R9810) and obtained abundance offspring materials which had stronger resistance to rice through cultivation, selection and collection.In order to show foreign DNA’s impact on genetic structure,we used the method of SSR molecular markers to analysis the genetic differences among Chaling wild rice,cultivated R9810receptor and offspring materials. The results showed that SSR primers which were associated with resistance to rice blast can be amplified corresponding bands from the chaling wild rice, while the receptor material not; And primer1has expanded the corresponding DNA stripes in offspring3,4,5,6,8,9,10,meanwhile the primer2was in offspring3,4,5,6,7,8,10. Offspring materials had obvious changes in rice blast resistant genetic characteristics compared to cultivated R9810receptor, which provided theoretical basis for successful import of the total DNA from Chaling wild rice.According to the known broad-spectrum resistance gene to rice blast, we respectively amplified7301bp and7294bp gene sequences which separately had92.49%and92.74%homology with Pi-ta gene (GeneBank login number:GQ918485.1)from Chaling wild rice and offspring5by using homologous cloning method;Analysis showed that the amplified gene was likely to be a new gene resistant to rice blast and the99.5%similarity between them fully demonstrated rice blast resistance gene in chaling wild rice has been transferred to common cultivated rice effectively.With D6import offspring of chaling wild rice ys090026.6, ys090036.1, ys09004/6as materials,to identify the rice blast resistance of seedling period, experimental results showed that ys090026.6,ys090036.1,ys09004/6all had strong disease resistance, including ys090026.6showed strongest resistance characteristic.These studies in germplasm resources of chaling wild rice provided important theoretical basis for the future breeding work in rice blast resistance and also provided technical support for the development and utilization of chaling wild rice.