Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23496
標題: Xanthomonas campestris pv. campestris第三型致病蛋白XopX8004的致病機制探討
Functional characterizations of Xanthomons campestris pv. campestris type III effector - XopX8004
作者: 楊珮筠
Yang, Pei-Yun
關鍵字: 十字花科植物
Xanthomonas campestris pv.campestris
黑腐病
第三型致病蛋白
black rot
type III effectors
出版社: 生物化學研究所
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摘要: Xanthomonas campestris pv.campestris是一種革蘭氏陰性植物病原菌,會造成十字花科植物感染黑腐病,而X. campestris pv.campestris是透過第三型分泌系統將第三型致病蛋白注射至宿主細胞。其中包括能抑制宿主免疫反應,使病原菌可以在宿主內增生之致病性以及宿主對於病原菌具有抗性,可以引發免疫反應之非致病性。目前為止,X. campestris pv.campestris已經超過25個致病蛋白被發現,但是這些致病蛋白的分子機制大多還不清楚。本實驗的主要目的是了解致病蛋白的分子功能,首先透過製備三個致病蛋白XopX8004、XopAC8004和XopP8004之抗體,以利偵測轉基因植物之蛋白表現以及在X. campestris pv.campestris 8004中之表現情形。且進一步針對XopX8004之功能做分析,其中透過雌激素誘導轉基因植物XVE::XopX8004可以發現其產生和能引發過敏性反應之XVE::AvrXccC8004 (110-440a.a.)轉基因植物有相似之病徵,且透過即時定量聚合酶連鎖反應偵測受到雌激素誘導之XVE::XopX8004-SFP能使植物免疫相關基因上升引起植物之免疫反應。另一方面透過次世代定序分析之結果觀察到,XopX8004能夠使植物防禦反應相關基因、防禦蛋白相關基因以及細胞死亡相關基因表現量提升,認為XopX8004在擬南芥中屬於非致病性蛋白。為了進一步了解XopX8004的致病機轉我們建構帶有並列式親和性純化標籤之XVE::XopX8004 -SFP轉基因植物,藉由親和性純化的方式。目前已經可以純化出植物細胞表現的XopX8004- SFP蛋白,期望可透過蛋白質體學的方式來尋找植物細胞中與XopX8004有交互作用的蛋白。
Xanthomonas campestris pv.campestris is a gram-negative phytopathogenic bacteria, which can cause the black rot of cruciferous plants. Injection of type III effectors into plant cells depends on the type III secretion system of X. campestris pv.campestris.Virulence effectors can suppress the host immune response and enable pathogen to grow within the host. Avirlence effectors can be recognized by the host resistance proteins and trigger immune responses. In X. campestris pv.campestris, more than 25 effectors have been identified but the molecular basis of pathogensis of type III effectors is still unknown. To better understand the molecular mechanism of pathogensis of Xcc type III effectors, anti-bodies against XopX8004, XopAC8004 and XopP8004 were prepared to detect the protein expression on transgenic plants, and to examine whether these effectors can be secreted from X. campestris pv.campestris 8004. We further explore the function of type III effector XopX8004. Here, transgenic Arabidopsis XVE::XopX8004 treated with β-estradiol showed similar phenotypes to XVE::AvrXccC8004(110-440a.a.) transgenic plant, which can trigger plant defense. RNA-seq analysis revealed that XopX8004 can induce the expression of plant immunity-related genes, resistance related genes and cell death related genes. These results suggest that XopX8004 behaves as an aviruelnce effector in Arabidopsis. In order to better understand the molecular action of XopX8004 in plant cells, a tandem affinity-tagged XVE::XopX8004 –SFP transgenic line was generated. Using strep-tactin resins, in planta-expressed XopX8004 was successful purified from plant. This approach was useful to identify XopX8004-interacting proteins.
URI: http://hdl.handle.net/11455/23496
其他識別: U0005-2208201315551300
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2208201315551300
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