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Rice stripe virus (Rice stripe virus, RSV) caused by rice stripe disease is the role of a rice production on a wide range of serious harm viral disease. RNA-mediated virus resistance is a post-transcriptional gene silencing (Post Transcriptional Gene Silencing, PTGS). The latest studies have shown that occur in different species of RNA silencing factors directly lead to double-stranded RNA (double Strand RNA). Transgenic Plants, the dsRNA formed an effective way is constructed in vitro hpRNA structure (hairpin RNA) cDNA introduced plants. Constructed in this study four RSV containing different functional genes hairpin RNA (hpRNA), the structure of the plant expression vector to form a double-stranded RNA (transcription), Agrobacterium-mediated transformation of rice, rice stripe virus resistance of Transgenic Plants identification, comparison RSV coat protein gene CP, specific gene SP, non-specific protein gene NS3 and sports-related protein gene NSvc4 differences in RNA-mediated virus resistance. The specific results are as follows: RSV CP the NS3, the SP and NSvc4 gene 3 'end of the 400bp fragment purpose respectively inserted into the inverted repeat comprising after the transformation of the plant expression vector the pCAMBIA - 1300' to construct a hairpin structure Plants to expression vector the pPHD-CP pPHD-NS3, pPHD-SP and pPHD-NSvc4 will build plant expression the the the carrier pPHD-CP, pPHD,-NS3, pPHD-SP and pPHD-NSvc4 and empty vector pCAMBIA1300 'freeze-thaw was introduced into Agrobacterium EHA105. Broth PCR and restriction enzyme digestion that plasmids have been successfully introduced into Agrobacterium correctly. 3, Agrobacterium-mediated transformation of rice varieties Yujing 6. By kanamycin resistance screening of regenerated plants identified by PCR, respectively CP rice turn pPHD-68 70 turn pPHD-NS3 rice, 57 the SP turn pPHD-rice, turn pPHD- 58 of the NSvc4 rice and turn pCAMBIA-1300 'of 75 of rice. 4, striatellus pass drug mediator, conventional biological inoculation, the rice receptor and resistance analysis. Statistical measurement results, in turn pPHD-CP pPHD-NS3, pPHD-SP and pPHD-NSvc4 rice, the proportion of RSV-resistant plants were 32.73%, 30.00%, 38.56% and 31.67%. 5, Northern blot and siRNA blot analysis results, transgenic RNA level expression, the amount of transcription product of the target fragment accumulation and transgenic plants disease resistance was negatively correlated; transgenic plants can be detected to the virus-specific siRNA, prove The resistance is RNA-mediated disease resistance, that RNA silencing results, but there is no direct relationship between the two resistance with siRNA accumulation.
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