計畫目標: 收集台灣各地的灰黴病菌株, 進行專一性核酸探針的選殖與專一性的評估, 以開發偵測與診斷用試劑, 並建立灰黴病指紋圖譜資料庫以供日後鑑定及應用發展之所需.架構: 為完成此目標首先必須收集灰黴病菌菌株與賴俊枝種苗種球, 進行菌株之分離及鑑定, 而後進行專一性核酸片段之選殖與進行專一性核酸探針之設計, 並利用逆點漬雜合技術以確定專一性核酸探針之專一性.同時發展應用於植物體上之檢測技術與檢測流程的建立.並以隨機增幅多型性分析( RAPD )及核酸限制酵素剪切片段多型性分析( RFLP )等技術建立灰黴病菌之核酸指紋圖譜, 以作為日後鑑定灰黴病菌之依據.預期效益: 完成以上的工作預計將可藉由對灰黴病菌生理生態的瞭解而掌控防治的時機以減少經濟損失, 並藉由專一性核酸探針的開發與標準檢測流程的建立將可加速對灰黴病菌的診斷與偵測速度.並藉由指紋圖譜的建立可作為鑑定灰黴病菌之依據與新菌系產生之評估指標.For the purpose of develop detection and diagnosis kits of Botrytis spp.First we collect the Botrytis isolates from Taiwan, and clone the specific DNA fragment for design the specific primer for detection of Botrytis spp.Use reverse dot blot hybridization to sure the specificity of the primer, and establish the process for detection and diagnosis.At same time we construct the fingerprint database of Botrytis spp.for the bases of identification and estimate for the occurrence of new strain of Botrytis spp.After we finished these work.We can know the time to control the disease caused by Botrytis spp.by understanding the physical and the ecology of Botrytis spp., and decrease the economical lose caused by Botrytis spp.By the development of the detection and diagnosis kits we can detect and diagnose Botrytis spp.faster.For the construction the database of Botrytis fingerprint we can use it for identify and estimate for the occurrence of new strain of Botrytis spp
