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題名 | 使用Real-time PCR檢測引起西瓜蔓枯病菌的方法評估=Evaluation of Real-time PCR Assay for Detection of Didymella bryoniae |
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作者 | 關政平; 黃彥華; 曾清山; 林宗俊; 石信德; Kuan, Cheng-ping; Huang, Yen-hua; Tseng, Ching-shan; Lin, Tsung-chun; Shih, Hsin-der; |
期刊 | 臺灣農業研究 |
出版日期 | 20140600 |
卷期 | 63:2 2014.06[民103.06] |
頁次 | 頁151-158 |
分類號 | 435.264 |
語文 | chi |
關鍵詞 | 西瓜蔓枯病; 診斷技術; Gummy stem blight; Didymella bryoniae; Real-time PCR; Detection; |
中文摘要 | 本研究根據西瓜蔓枯病菌(Didymella bryoniae)的rDNA設計出專一性的引子對及利用real-time PCR法與TaqMan螢光探針進行D. bryoniae之檢測並與傳統PCR法作比較,結果顯示real-time PCR法較傳統PCR法靈敏度提高約100倍。利用real-time PCR法偵測被D. bryoniae在感染的西瓜組織,結果發現D. bryoniae主要分布在西瓜的莖部。本研究使用real-time PCR法可增幅出D. bryoniae的專一性核酸片段,並檢測感染西瓜組織及種子中之D. bryoniae核酸,將可提供西瓜蔓枯病之病害檢測與病菌量化之判定。 |
英文摘要 | In this study, a specific and highly sensitive real-time PCR assay with TaqMan probe was developed as a tool for the diagnosis of gummy stem blight of watermelon by targeting the rDNA genes of "Didymella bryoniae." In addition, the real-time PCR assay was compared to a conventional PCR method with their specificity and sensitivity on the detection of "D. bryoniae" in culture andin planta. The real-time PCR assay was effective for quantifying the fungal genomic DNA in the infected plants. This method was highly specific to "Didymella" and it was about one hundred times more sensitive than the conventional PCR method. The real-time PCR assaydeveloped isavaluable tool for detection and titer quantitation of gummy stem blight diseases on watermelon. |
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