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Molecular Detection of Fumonisin Producing Fusarium Species of Freshly Harvested Maize Kernels Using Polymerase Chain Reaction (PCR)

利用聚合酶連鎖反應(PCR)偵測從新鮮收成玉米粒分離出來的會產生Fumonisin的鐮胞菌株

摘要


Fumonisins是新種類的有毒二次代謝物,由鐮胞菌(Fusarium species) Fusarium moniliforme和Fusarium proliferatum產生,這兩種鐮胞菌近來被科學界廣泛研究。從新鮮收成玉米粒分離出來32株Fusarium,分別是Fusarium moniliforme(12株),Fusarium proliferatum(5株),Fusarium anthophilum(4株),Fusarium oxysporum(3株),Fusarium sporotrichioides(3株),Fusarium pallidoroseum(3株)和Fusarium solani(2株),以及Aspergillus flavus(2株)和Alternaria alternate(2株)都用來進行PCR分析。本研究目標為發展出一套聚合酶連鎖反應(PCR)偵測系統,用以偵測產生Fumonisin的鐮胞菌株。一組參照鐮胞菌屬具保留性的ITS區域設計的核酸引子(primer)用來區分鐮胞菌屬和其他真菌菌屬,例如Aspergillus和Alternaria。另一組參照鐮胞菌產生fumonisin的“FUM 1基因”區所設計的核酸引子用來區分產生fumonisin的鐮胞菌種和不產生fumonisin的鐮胞菌種。研究結果顯示,利用PCR技術不僅可以用來區分鐮胞菌屬和其他真菌菌屬,也可以用來鑑定產生fumonisin的Fusarium moniliforme,Fusarium proliferatum和Fusarium anthophilum。

並列摘要


Fumonisins are a new class of toxic secondary metabolites produced by Fusarium species namely Fusarium moniliforme and Fusarium proliferatum that have recently come under intense scientific investigations. Thirty-two Fusarium isolates namely, Fusarium moniliforme (12 isolates), Fusarium proliferatum (5 isolates), Fusarium anthophilum (4 isolates), Fusarium oxysporum (3 isolates), Fusarium sporotrichioides (3 isolates), Fusarium pallidoroseum (3 isolates) and Fusarium solani (2 isolates), other four isolates of Aspergillus flavus (2 isolates) and Alternaria alternata (2 isolates), isolated from freshly harvested maize kernels were subjected to PCR analysis. The objective of this study was to develop a PCR-based detection kit for detection of fumonisin-producing Fusarium species. One set of primers based on the conserved ITS region of the genus Fusarium was used to differentiate the genus Fusarium from other fungal genera such as Aspergillus and Alternaria. The other set of primers specific to fumonisin producing 'FUM 1gene' region of Fusarium was used to differentiate the fumonisin producing Fusarium species from non-fumonisin producers. Results of the study revealed that PCR-based technique could be used not only to differentiate the Fusarium species from other genera of fungi but also to identify fumonisin-producing Fusarium moniliforme, F. proliferatum and F. anthophilum.

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