Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/56119
標題: 功能性鑑定刺角瓜木瓜輪點病相關抗病基因
Functional Characterization of Prsv-Related Resistance Genes in Cucumis Metuliferus
作者: 古新梅
詹富智
關鍵字: 植物保護類
商品化
摘要: 許多瓜類在熱帶及亞熱帶地區容易受到木瓜輪點病毒(papaya ringspot virus, PRSV)之侵害,使植株生長受阻,造成產量及品質下降,因此了解植物抗病毒機制與獲得抗病之栽培品種為目前重要的工作。目前已知刺角瓜(Cucumis metuloferus )品系P.I. 292190(Line 37) 具有一單一顯性抗性基因 Wmv,可對抗PRSV。為了確立並分離出相關的抗性基因,先前研究利用cDNA-AFLP 及reverse Northern 分析後,已成功獲得24 個與抗病相關的候選cDNA 片段,也成功建立刺角瓜的再生與農桿菌轉殖系統。在此三年計畫裡擬利用RNAi 及Yest-two-hybrid 分析兩種策略來找出刺角瓜Wmv 基因及與PRSV 相關的抗病基因。首先在RNAi 分析方面,目前已成功構築六個候選基因於RNAi 表現載體中並轉殖入抗病品系Line 37,其中候選基因Cm4 經RNAi 試驗後證實可提供抗病功能,將進一步選殖全長基因序列並進行互補試驗。另外,目前研究推測病毒之HC-Pro蛋白可能與Line 37 抗病相關基因間具有交互作用,因此HC-Pro 蛋白及其突變型式與刺角瓜L37 之間抗感性的變異提供研究病毒與寄主交互作用非常理想的模式,故本計畫擬利用HC-Pro 及其突變型式蛋白與從刺角瓜L37 得到之後選基因之間進行Yeast-one-hybrid 分析,期望可分得到與HC-Pro 蛋白有交互作用的抗病基因,並了解寄主與病毒之共演化關係,最後本研究所得與抗PRSV 相關的刺角瓜候選基因將轉殖於菸草及其他瓜類進一步探討其在異源作物是否能提供PRSV 之抗性。
Papaya ringspot virus (PRSV) causes severe damages of crops especially papaya andcucurbits. Therefore, it is important to understand the virus resistant system of plants and tobreed the resistant variety. A single domain resistant gene, Wmv, in horned melon, Cucumismetuliferus Line P.I. 292190 (Line 37) was reported contributing strong resistance toPRSV-HA, which provides an idea model to investigate the underlying molecular mechanismsof host and virus interactions. In our previous studies, we have already used cDNA-AFLPand reverse Northern analysis to screen the candidates of resistant line L37 and susceptibleline L35 which differential expression against PRSV and obtained 24 candidate cDNAfragments. 6 of the 24 candidate fragments were already constructed on RNAi expressionvector, and one of the candidates, Cm4, was showed ability to confer the resistance againstPRSV. In this three-year proposal, we are going to use RNAi approach to evaluate thepotential resistant function of the cDNA candidates, cloning the full length cDNA fragmentsfor complementary test, and yeast two-hybrid system which will be performed to fish out thepotential PRSV-related resistance genes in L37 cDNA candidates specifically interacting toPRSV HC-Pro. Finally, these resistant-related genes will be agrobacterium-transformed andfunctional characterized in horened melon and tobacco. This will lead to further understandfull pictures of molecular mechanisms interacting between virus and plant host.
URI: http://hdl.handle.net/11455/56119
其他識別: NSC99-2313-B005-016-MY3
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=2224918&plan_no=NSC99-2313-B005-016-MY3&plan_year=100&projkey=PD10001-0053&target=plan&highStr=*&check=0&pnchDesc=%E5%8A%9F%E8%83%BD%E6%80%A7%E9%91%91%E5%AE%9A%E5%88%BA%E8%A7%92%E7%93%9C%E6%9C%A8%E7%93%9C%E8%BC%AA%E9%BB%9E%E7%97%85%E7%9B%B8%E9%97%9C%E6%8A%97%E7%97%85%E5%9F%BA%E5%9B%A0
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