Research Paper

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

M. Y. Sreenivasa, Regina Sharmila Dass, A. P. Charith Raj and G. R. Janardhana

Published on: December 2006

Page: 251 - 257

DOI: 10.6165/tai.2006.51(4).251

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    Abstract

    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.

    中文摘要

    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。

    Keyword: Fusarium species, Maize, Fumonisin-producers, Molecular detection technique, Polymerase Chain Reaction.

    Literature Cited