Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/56062
標題: 功能性鑑定刺角瓜抗木瓜輪點病毒相關抗病基因
Functional Characterization of Prsv-Related Resistance Genes in Cucumis Metuliferus
作者: 古新梅
詹富智
關鍵字: 基礎研究;Papaya ringspot virus(PRSV);植物保護類;木瓜輪點病毒;刺角瓜;北方墨點法;基因靜默;酵母菌雙雜交;Cucumis metuliferus;Northern blot;RNAi;yeast two-hybrid
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,先用北方墨點法分析Line 37 接種與未接種PRSV植株在不同time course 中具有差異表現的候選片段,並將這些candidates 利用RNAi 方式轉殖入抗病品系Line 37,探討候選基因在抗感病反應之角色,並藉此選殖出與抗PRSV 相關的基因。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 蛋白以及其突變型式與刺腳瓜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,故本計畫擬利用葉錫東教授提供的HC-Pro 及其突變型式蛋白與刺腳瓜L37 之間的抗感差異反應進行Yeast-two-hybrid 系統分析,期望可分離出與抗病相關之基因,並了解寄主與病毒之共演化關係。

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. In previous researches, a single domain resistant gene, Wmv, inhorned melon, Cucumis metuliferus Line P.I. 292190 (Line 37) provides strong resistance toPRSV-P, 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. In this three-year proposal, we are going to use time-course Northern analysis andRNAi approach to evaluate the potential resistant function of the cDNA candidates. On theother hand, yeast two-hybrid system will be performed to fish out the potential PRSV-relatedresistance genes in horned melon L37 which interacting to PRSV HC-Pro. Finally, theresistant gene Wmv and PRSV resistant related genes will be functional characterized andunderstood the molecular mechanisms interacting between virus and host resistant systems.
URI: http://hdl.handle.net/11455/56062
其他識別: NSC97-2313-B005-029
Appears in Collections:農藝學系

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