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Rice stripe virus (Rice stripe virus, RSV) the slender virus, (Tenuivirus), a typical member of has been distributed in 16 provinces and cities in China, the outbreak is still in some provinces and cities in China, causing serious damage to China's rice production. Fujian Agriculture and Forestry University, where the virus RSV biology, pathology and molecular biology done a lot of research work, and speculated that the coat protein of RSV (coat protein, CP) and disease-specific protein (disease-specific protein SP) for possible disease-causing proteins. In this paper, transgenic technology, the CP and SP genes were transformed into rice, in order to further study the function of these two proteins. Total nucleic acid extracted from the diseased leaves of infestation the RSV Beijing Shuangqiaoshan, isolates (RSV-SQ), to obtain CP, SP gene fragment was amplified by RT-PCR, and cloned into pGEM-T easy vector, to obtain a recombinant plasmid pTE -CP and pTE-SP; via BamH I / Sac I double digested recombinant plasmid and the corresponding fragment of the purpose of recycling, to its connection to the same double digested plant expression vector pCAMBIA2301; and then double digested by EcoR Ⅰ and Hind Ⅲ, will cut under containing 35Spromoter, target fragment, termination of the NOS-ter non-coding sequences ligated to the expression vector pCAMBIA1300, plant expression vector the pCA1300-CP and pCA1300-SP. And by direct DNA transformation, power transformation and tri-parental mating transferred to Agrobacterium. The results show that the efficiency is better than the first two methods of transforming DNA tri-parental mating; mediated by Agrobacterium tumefaciens CP, SP gene were transformed into TNG 67 rice varieties. After screening, the regeneration of the transformed plants. Point PCR and Southern blot analysis preliminary findings show that the target gene fragment were integrated into the rice genome. PCR-Southern dot blot results showed that the part of the target fragment is transcribed in plant. Preliminary observations indicate that RSV-CP, SP Gene Transformation of Rice alone does not cause symptoms similar to virus infection, indicating that the CP and SP genes alone can not pathogenic. To further explore the molecular evolutionary relationships of SP gene function and virus isolated from pathogenic the RNA4 fragment of the RSV-SQ sequences were analyzed. The results showed that the the RNA4 total length of 2157bp. Sequence homology at the nucleotide level, RSV-SQ Panjin (RSV-PJ), Japan T isolates (RSV-T), M isolates (RSV-M) and Chuxiong isolates ( the RSV-CX) vRNA4 of ORF homology were 98.1%, 98.3%, 98.3% and 96.1%, the homologies were vcRNA4 ORF was 98.4%, 98.1%, 97.8% and 94.0%, homology RNA4 IR were 94.0%, 94.8%, 89.4% and 85.4%; homology at the amino acid level, vRNA4 ORF Fujian Agriculture and Forestry University, a master's degree thesis were 99.4%, 100%, 98.9% and 98.9% , VCRNA4 ORF sequence homologies were 99.00 and 97.60. Found in the the RNA4 full-length sequence homology comparison RSVS with RSV. PJ. RSV. T, RSV-M, RSV-CX homology, respectively, for 96%, 96%, 95%, 90%;-PJ and RSV RSV. T, RSV-M, RSV-CX homology, respectively, for 96%, 95%, gi%; RSV-T with RSV-M, RSV-CX homology of 95% and 91%, respectively; RSV- M with RSV-CX, the homology was 92%. Thus, RSVSQ with RSV-CX, although both belong to the Chinese sub. From the material, but the kinship more RSVPJ and RSV-T, RSV-M is far. Sequence analysis also found that five isolates 5 UTR sequence identical to the 3 UTR only RSV-CX isolate two nuclear celecoxib acid changes, and in RNA4 IR, containing A, U base enriched with inverted repeats can form a hairpin structure. Variation in each section were discussed.
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