2024-03-26
【學術亮點】開發蕪菁尖鐮孢菌及紫羅蘭尖鐮孢菌專一性引子對並整合多重PCR用於區分十字花科植物上的四個分化型病原菌
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Intelligent Detection: Development of expert diagnosis system for crop cultivation and management【Department of Plant Pathology / Chih-Li Wang / Associate Professor】
智慧檢測：作物栽培管理專家診斷系統開發【植病系王智立副教授】
上架日期2023-12-21
智慧檢測：作物栽培管理專家診斷系統開發【植病系王智立副教授】
| 論文篇名 | 英文：Development of specific primers for Fusarium oxysporum formae speciales rapae and matthiolae with an integrated multiplex PCR for distinguishing four formae speciales on Brassicaceae 中文：開發蕪菁尖鐮孢菌及紫羅蘭尖鐮孢菌專一性引子對並整合多重PCR用於區分十字花科植物上的四個分化型病原菌 |
| 期刊名稱 | Plant Disease |
| 發表年份, 卷數, 起迄頁數 | 2023(Online) |
| 作者 | Huang-Hsi Chu, Wei-Chin Tsao, Jenn-wen Huang, Pi-Fang Linda Chang, Chih-Li Wang (王智立)* |
| DOI | 10.1094/PDIS-08-23-1656-RE |
| 中文摘要 | Fusarium oxysporum是造成十字花科作物黃葉病的病原菌，目前世界上共有4種分化型(formae speciales)，由於十字花科作物群間的交互育種，不同分化型病原菌的寄主範圍時常重疊，使鑑定病原菌成為一大挑戰。其中蕪菁尖鐮孢菌(F. oxysporum f. sp. rapae)及紫羅蘭尖鐮孢菌(F. oxysporum f. sp. matthiolae) 目前仍舊沒有專一性引子對可供鑑定，為了解決這個問題，本研究鎖定病原性F. oxysporum特有的效應蛋白“SIX 基因”進行引子對的設計。透過與其他分化型菌株的序列比對，成功設計蕪菁尖鐮孢菌及紫羅蘭尖鐮孢菌專一性引子對分別於SIX14及SIX9上，兩組引子對展現了高度的專一性於偵測目標菌株，並且可以從20種非目標分化型、5種Fusarium屬病原菌及4種十字花科常見病原真菌中區分。此外，這些專一性引子對的有效性通過檢測感染植物中的病原體而得到驗證。為進一步增強對四個分化型病原菌的鑑定步驟，我們結合了兩對新設計的專一性引子對與兩對前人研究發表的引子對，建立了一種多重PCR方法，可以在單一反應中準確區分引起十字花科植物黃葉病的Fusarium oxysporum四個分化型。 |
| 英文摘要 | There are four formae speciales of Fusarium oxysporum responsible for causing yellows of Brassicaceae. Due to cross-breeding among crops, the host ranges of these formae speciales often overlap, making pathogen identification a challenging task. Among these formae speciales, F. oxysporum f. sp. rapae and F. oxysporum f. sp. matthiolae still lack specific primers for pathogen identification. To address this problem, we targeted the SIX genes, known as specific effectors of pathogenic F. oxysporum, for primer design. Through sequence comparison with other formae speciales, we successfully designed specific primers for F. oxysporum f. sp. rapae and F. oxysporum f. sp. matthiolae on SIX14 and SIX9, respectively. Both primer pairs exhibited high specificity in detecting F. oxysporum f. sp. rapae or F. oxysporum f. sp. matthiolae, distinguishing them from 20 non-target formae speciales of F. oxysporum, five species of phytopathogenic Fusarium, and four other common pathogenic fungi affecting cruciferous plants. Moreover, the effectiveness of these specific primers was validated by detecting the pathogens in infected plants. To further enhance the identification process of the four formae speciales, we combined the two newly designed specific primer pairs with two previously published primer pairs, enabling the establishment of a multiplex PCR method that can accurately distinguish all four formae speciales of F. oxysporum responsible for causing yellows in cruciferous plants in a single reaction. |
| 發表成果與AI計畫研究主題相關性 | 為建立未來AI樣本及照片資料庫，採樣發表成果相關病害進行病害照片拍攝，並將樣本帶回實驗室檢測及驗證確認病原菌。 |
