草莓镶脉病毒致病相关基因功能分析-植物病理学专业论文.docxVIP

II II 白可能对P6蛋白有一定的负调控作用；P4蛋白和P6蛋白的互作显著加重了症状， 表现出极强的致病性，说明P4蛋白和P6蛋白之间可能具有一定的协生作用。 4. SVBV P6基因在本氏烟中稳定表达的症状表型 将构建的重组质粒pCAM-P6-US转化农杆菌EHA105，叶盘法转化本氏烟。 抗性筛选及提取转基因烟草总DNA，特异性引物PCR扩增后发现，目的基因已成 功整合进本氏烟基因组中。症状观察发现，P6转基因本氏烟没有表现出特殊的 症状表型。 5. P6 与植物抗病性相关基因 06G 的互作 构建重组质粒 pCAM-P6-US、pCAM-2b 和 pCAM-HC-Pro，转化农杆菌分别 浸润本氏烟，6 d 后采集浸润叶片提取总 RNA，Real-Time PCR 检测植物抗性相 关基因 06G 的 RNA 积累水平。结果发现，含有 pCAM-P6-US 的农杆菌浸润本 氏烟可以诱导寄主 06G 基因 RNA 的高水平积累。 关键词：草莓镶脉病毒；致病相关蛋白；瞬时浸润；稳定表达；症状表型；植物 抗病基因 06G PAGE PAGE IV Abstract Corresponding symptom phenotypes usually appears after hosts being infected with viruses, and the phenotype probably has some relationship with pathogenic proteins encoded by virus. It was demonstrated that P6 protein encoded by Strawberry vein banding virus was a weak RNA silencing suppressor. On these bases, further research work about pathogenicity of P6 and other proteins encode by SVBV was carried out. Not only the pathogenicity of a solo protein encoded by SVBV was identified, but also the interaction of the proteins of SVBV was conducted. In addition, whether P6 protein has influence on the expression of gene 06G related to the plant resistance was carried out preliminaryly, and it established the foundation for illuminating the pathogenic molecular mechanism in our work. Cloning and Construction vectors of pathogenicity related genes of SVBV Gene P1, P2, P4, P6 and P6 frameshift mutation were amplified, using the full-length plasmid of SVBV American isolate as template (pSVBV-E3), then these were inserted into the viral vector pGR106, respectively. The infectious clone recombinant vectors pGR-P1-US, pGR-P2-US, pGR-P4-US, pGR-P6-US and pGR- △P6-US were obtained. Moreover, gene P6 was also constructed into the binary expression vector pCAM2300 and the recombinant expression vector pCAM-P6-US was obtained. Then, each of recombinant vectors was transformed into Agrobacterium tumefaciens GV3101. Pathogenicity related proteins enconded by SVBV were verified by trasient infiltration