目录
摘要II
关键词II
AbstractIII
引言
引言1
1材料与方法4
1.1供试材料4
1.1.1供试菌株4
1.1.2供试植物4
1.1.3常用培养基4
1.2农杆菌介导的瞬时表达5
1.3寄生疫霉接种本氏烟5
1.4活性氧测定6
1.5敲除转化子制备6
1.5.1构建敲除载体6
1.5.2电击转化青枯菌7
1.5.3交换子验证7
2结果与分析8
2.1 Rip25能够诱导本氏烟细胞死亡8
2.2 Rip25不影响寄生疫霉侵染本氏烟8
2.3 Rip25能够抑制flg22诱导的活性氧迸发9
2.4敲除转化子构建10
2.4.1敲除载体的构建10
2.4.2电击转化青枯菌11
3讨论12
致谢13
参考文献13
青枯菌III型效应子Rip25鉴定与功能初探
摘 要
青枯菌(Ralstonia solanacearum)是世界上最具破坏性的植物病原细菌之一，是一种典型的土壤传播病原菌。其寄主范围广泛，除危害番茄外，还可危害马铃薯、茄子、辣椒等50多个科200余种重要作物，引发植物青枯病，给农业生产带来巨大的威胁。青枯菌是一种革兰氏阴性细菌，通过III型分泌系统向寄主细胞分泌大量的III型效应子，干扰植物的免疫，促进侵染，其效应子数目众多，但目前仅有少数几个效应子功能明确。本研究鉴定了青枯菌模式菌株GMI1000的III型效应子Rip25，利用农杆菌介导的瞬时表达系统在本氏烟上瞬时表达该效应子，发现Rip25能诱导本氏烟细胞死亡，抑制细菌鞭毛蛋白flg22诱导的活性氧迸发，但不能影响寄生疫霉侵染本氏烟。课题计划结合基因敲除的方法，对Rip25的功能进行初步分析，有望为开发新型青枯病防控策略提供理论依据。
FUNCTIONAL CHARACTERIZATION OF RALSTONIA SOLANACEARUM TYP *51今日免费论文网|www.51jrft.com +Q: #351916072# 
E III EFFECTOR RIP25
ABSTRACT
Ralstonia solanacerum is a typical soilborn pathogen and one of the most destructive plant pathogenic bacteria in the world. It has a huge host range with more than 50 species and 200 families of plants, including tomatoes, potatoes, eggplants, peppers and so on. It is the cause of plant bacterial wilt, which is a huge treat to agricultural production. Ralstonia solanacerum is a gramnegative bacterium that can secrete a large number of type III effectors into host plant cells through type III secretion system, interfering plant immune system and promoting its infection. Ralstonia solanacerum has many effectors, but only a few of them are functional understood. This study focuses on the type III effector Rip25 of GMI1000, and mainly uses agrobacteriummediated transient expression technology with model plant N. benthamiana. The results show that Rip25 can induce tobacco cell death, inhibit the burst of ROS, but has no significant effect on the disease resistance of tobacco. Based on the research results above, along with the geneknockout method, we hope to refer the function and mechanism of Rip25 in the process of Ralstonia solanacerum infecting plants and provide a theoretical basis for establishing a new type of Ralstonia solanacearum disease control strategy.
KEY WORDS：Ralstonia solanacearum; type III effectors; transient expression; gene knockout

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