Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/53405
標題: 農桿菌腫瘤蛋白6b及6b結合蛋白在調節植物防禦反應上的功能性分析
Functional Analysis of Agrobacterium Oncoprotein, 6b, and 6b-Interacting Proteins in the Regulation of Plant Defense Response
作者: 楊俊逸
關鍵字: 生物科學類
基礎研究
Agrobacterium
geminivirus
jasmonic acid
6b
bC1
AS1
SE
摘要: 者度增一 , 上加個在例,會廣自使如泛改然我病變傳界們原寄播,更菌主的植加和植的物了食物反會解草病應和植昆,原各物進蟲菌種如而。,生何昆影可物協蟲響以有調是到藉交和病後由互整原來腫作合菌入瘤用他感侵誘,們者染導因植昆的此物蟲防植的禦的物媒生反可介存應以。。來成近當對為來病抗各這原各種種菌種生三感生物方染物間互植的交動物威互的後脅作研,。用究在農的某不桿中種斷菌介在是程DNA (T-DNA)腫的瘤退化在植瓦物解,上進形而成促後使,腫會瘤成中為的其農他桿病菌原釋菌回感土染壤和中昆。蟲目入前侵我的們點的,的初而轉步這移研種來究感引顯染起示及植,入物農侵的桿會腫菌導瘤致病腫。瘤T-DNA中所帶有的6b蛋白,可能具有類似於雙生病毒bC1蛋白的功能。bC1的一個致病因子。是番茄黃化曲葉病毒中bC1已被證實會和擬南芥中調控葉片極性和免疫反應的AS1而影響葉子發育和抑制茉莉酸蛋白結合,進(jasmonic acid)反應。我們初步的實驗結果不僅提供6b和AS1互作用的證據,而且還找到新的交6b標的蛋白SERRATE (簡稱SE)。SE是一個參與mRNA接和剪miRNA加工的鋅指(zinc-finger)蛋白。因此我們推測6b藉由同時與AS1及SE來嚴格調控的交互作用,6b他和病分原子生菌物和學昆蟲的對方的於法感植,染物來及茉分入莉析侵酸擬。反南本應芥研的究抑計制畫功將能集,中而於讓下農列桿的菌幾感個染方所向引來起進的行植探物討腫:瘤1.更利易用受遺其傳6b轉殖株的茉莉酸反應。2.利用in vivo及in vitro作用實驗,來研究的交互6b-AS1和6b-SE之間的交互作用和確定交互作用的蛋白區塊。3.分析6b是否具有抑制後轉錄基因沉寂的活性。4.利用染色質免疫沉澱法(ChIP assay),來分析SE物免疫反應中所扮演的角色。5.利用酵母雙雜交在植(yeast two-hybrid)與茉莉酸反應的篩選和遺傳分析,來鑑定參SE結合蛋白。此研究結果將有助於在植物中發現新的病原菌標的物,進而對植物的細胞機制提供嶄新的觀點,以期衍生出有效的技術來進行植物保護。
In nature, plants interact with a range of organisms; in such interactions plants serve as a mediator,for example, between microbial pathogens and insect herbivores. The infection by pathogenssomehow can alter the host plants in ways that affect the survival of subsequent attacker, insectvectors. Research studies in such tripartite interactions have increased for understanding how plantscoordinate and integrate their defenses against diverse threats. Agrobacterium tumefaciens, awild-spread phytopathogen, can cause crown gall disease by injecting a tumor-inducing transferredDNA into plant cell. Crown galls are sites to be infection by other pathogens and insects leading todegradation of crown galls and release of A. tumefaciens back into the soil. Our preliminary datashows that 6b, an oncoprotein encoded by transferred DNA, may act similarly as geminivirus bC1, apathogenicity factor of Tomato yellow leaf curl china virus. bC1 has been demonstrated to alter leafdevelopment and suppress jasmonic acid responses by interacting with Arabidopsis AS1, a conservedregulator for leaf polarity and immune response. The preliminary assays not only provide interactionevidences for 6b and AS1but also lead us to identify new host target, SERRATE (SE), a zinc-fingerprotein involving in pre-mRNA splicing and miRNA processing. It appears that 6b may suppressjasmonic acid responses by interacting simultaneously with AS1 and SE to bring about a tighterregulation. The suppression of jasmonic acid responses may in turn to cause the crown galls to bemore susceptible to infection. This proposal will focus on the following directions: 1. Characterizationof jasmonic acid responses in 6b transgenic plants using genetic and molecular approaches. 2. In vivoand in vitro interaction studies to dissect the interactions and define the interacting domains of 6b-AS1and 6b-SE. 3. Characterization of RNA silencing suppression activity of 6b. 4. Chromatinimmunoprecipitation (ChIP) analysis to dissect the functions of SE in plant immune responses. 5.Identification of SE interacting protein involving in jasmonic acid responses using yeast-two hybridscreening and genetic analysis. This line of investigation will lead to identify new host targetsinvolving in tripartite interaction and uncover novel aspects of plant fundamental cellular mechanisms,which may generate new technologies useful for plant protection.
URI: http://hdl.handle.net/11455/53405
其他識別: NSC99-2311-B005-002-MY3
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=2131037&plan_no=NSC99-2311-B005-002-MY3&plan_year=99&projkey=PA9907-2812&target=plan&highStr=*&check=0&pnchDesc=%E8%BE%B2%E6%A1%BF%E8%8F%8C%E8%85%AB%E7%98%A4%E8%9B%8B%E7%99%BD6b%E5%8F%8A6b%E7%B5%90%E5%90%88%E8%9B%8B%E7%99%BD%E5%9C%A8%E8%AA%BF%E7%AF%80%E6%A4%8D%E7%89%A9%E9%98%B2%E7%A6%A6%E5%8F%8D%E6%87%89%E4%B8%8A%E7%9A%84%E5%8A%9F%E8%83%BD%E6%80%A7%E5%88%86%E6%9E%90
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