| The disease caused by rice stripe virus is one of the most important viral diseases, and it is a threat to rice yield in our country. According to the theory of miRNA, several artificial microRNAs (amiRNAs) were designed for down-regulating the gene expression of RSV. The precursor of miRNA528 was cloned and used for construction of 2 amiRNA genes, amiR-1 and amiR-2, by over-lapping PCR. These genes were ligated into plasmid pBI121 respectively, The pAmiR-1 and pAmiR-2 are specific for interfering the expression of p3 and pc3 genes encoded by RSV RNA3. The 2 vectors were transferred into the japonica rice cultivar Wuyujing 3 by Agrobacterium-mediated transformation.219 transgenic rice plants (To) harbouring the amiRNAs were identified by PCR. The T1 transgenic plants were used to evaluate the resistance to rice stripe virus by artificial inoculation. The results showed that 22 plants are highly resistant to RSV. Real-time RT-PCR analysis suggested the transgenic amiRNA could be expressed at high level expressed in transgenic plants.
|
PDF Full Text Request