Journal of Food, Agriculture and Environment

A multiplex PCR assay for genetic chemotyping of toxigenic Fusarium graminearum and wheat grains for 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol and nivalenol mycotoxin

Author(s):

Jian-Hua Wang ¹, Jing-Bo Zhang ^{1, 2}, Fang-Fang Chen ¹, He-Ping Li ^{1, 3}, Mbacke Ndoye ¹, Yu-Cai Liao ^{1, 2}*

Recieved Date: 2012-01-02, Accepted Date: 2012-04-28

Abstract:

Fusarium graminearum causes Fusarium head blight (FHB) of wheat and other cereals and produces various types of trichothecene mycotoxins that are toxic to humans and domestic animals. Type B trichothecene mycotoxins, including deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON) and nivalenol (NIV), are the principal mycotoxins produced by toxigenic F. graminearum and the predominant mycotoxin contaminants in food/feed in China and many other countries. Rapid, accurate detection of the mycotoxins is essential to preventing them from entering food/feed chains. Based on the single nucleotide polymorphisms present within the Tri11 gene sequences involved in the trichothecene mycotoxin biosynthesis, a multiplex PCR assay was developed to simultaneously detect a 334-bp fragment from 3-AcDON–producing strains, a 279-bp fragment from 15-AcDON producers and a 497-bp fragment from NIV producers, which was completely congruent with chemical analyses of those mycotoxins that were extracted from wheat grains infected by 14 different mycotoxin-producing strains from barley, corn and wheat in China and from mycelium of one Swedish strain from wheat. This method was extended to assay 46 Fusarium strains from different hosts in China, Nepal, the United States and Europe, with the results confirming its global applicability and reliability. Moreover, the multiplex PCR was able to efficiently accomplish genetic chemotyping of wheat grains artificially infected by known chemotype-producing strains, indicating an identical chemotype pattern as the strains produced. Further application for testing naturally infested wheat grains collected from the fields in 14 different regions revealed simultaneous detection of three chemotypes from four samples and two chemotypes from eight samples, with only two samples having one chemotype. 3-AcDON chemotype was detected in 13 of the 14 wheat samples, which is well consistent with 3-AcDON producers being the predominant Fusarium strains on wheat in these regions. Sequencing analyses confirmed that these PCR-amplified fragments from wheat grains were indeed derived from the Tri11 gene sequences of F. graminearum and the chemotype-specific DNA fragments. Taken together, the Tri11 gene–based multiplex PCR assay is a rapid, reliable and globally applicable method, and it can be used for genetically predicting mycotoxin chemotypes of both toxigenic fungal strains and infested cereal grains in food/feed or agriculture samples.

Keywords:

Fusarium graminearum sensu lato, genetic chemotyping, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, wheat grain

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Journal: Journal of Food, Agriculture and Environment
Year: 2012
Volume: 10
Issue: 2
Category: Agriculture
Pages: 505-511

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