辣椒多聚半乳糖醛酸酶抑制蛋白基因克隆及功能研究-植物病理学专业论文.docxVIP

辣椒多聚半乳糖醛酸酶抑制蛋白基因克隆及功能研究 辣椒多聚半乳糖醛酸酶抑制蛋白基因克隆及功能研究 4 4 four-six-leaf stage pepper were treated with stress factors, and quantitative PCR analysis show that the three genes all exibit upregulated expression patteren against different stress factors, which indicated that PGIP involved in resistance signal transduction of plants. The inhibitory effects of three purified CaPGIP protein to several PGs. The CaPGIP protein were expressed by prokaryotic expression system and purified by Ni-NTA affinity chromatography. We collected several plant fungi and oomycetes pathogens and got their cell wall degrading enzymes (including PGs) induced by pectin. The inhibitory activity of CaPGIPs to PGs detected by UV spectrophotometry and agar diffusion show that the three CaPGIPs perform different inhibitory effects to PGs. The disease resistance analysis of pepper after CaPGIP gene silencing.The silencing vector about CaPGIP gene called pTRV2-CaPGIP was constructed, which will lead to CaPGIP genes silencing in part or whole through virus induced gene silencing (VIGS) mechanism, then quantitative PCR show that the expression of CaPGIP were decreased and inoculation detection reveal that the condition of pepper leveas against phytophthora capsic strain SD33 zoospore suspension were more sensitive than the controls. The disease resistance detection of transgenic tobacco. A transgenic plant expression vector pROKⅡ-CaPGIP1 of CaPGIP1 was constructed, the plasmid was transferred into tobacco to achieve a stable expression, and the T0 generation was obtained by plant regeneration. The positive plants were inoculated with Colletotrichum destructivum strain TBC-1 and Alternaria alternate strain TBA-003 after PCR testing, which showed stronger resistance than wild-types. So, CaPGIP show widely and specially inhibitory activity to PGs, the overexpression of PGIP does enhance the resistance of host, which provide foundation for carrying out PGIP transgenic overexpression in pepper to