Detection and tracing of Tri16 gene in T-2 toxigenic and nontoxigenic Fusarium species by PCR

Document Type : Original Research

Authors

1 Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

2 Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modarres University, Tehran, Iran

Abstract

  Aims: T-2 mycotoxin is the most toxic trichothecene produced by Fusarium species especially Fusarium sporotrichioides. This species is the contaminant of cereal grains that can cause severe diseases among humans and animals and it may lead to death. Tri16 gene encodes an acyltransferase that catalyzes the formation of ester side groups at C-8 in biosynthesis pathway of T-2 toxin. The aim of the present study was to design a molecular method for identification of Fusarium species based on Tri16 gene, capable of producing T-2 toxin.   Methods: In this experimental study, the DNA of 111 T-2 producing and nonproducing Fusarium strains (including 89 Fusarium isolates and 22 standard Fusarium spp.) were evaluated. After purifying the strains and preparing the cultures in Potato Dextrose Broth (PDB), the DNA was extracted using liquid Nitrogen and lysing buffers and by Phenol, Chloroform and Isoamyl alcohol (PCI) solutions. The gene was detected in all T-2 producing and nonproducing strains using the designed and selected primers based on Tri16 gene and PCR optimization.   Results: All T-2 toxin producing Fusarium strains including F usarium sporotrichioides (4 species), F usarium langsethiae (1 species) and F usarium Poae (1 species) contained the Tri16 gene fragment (1380 bp).   Conclusion: Important T-2 toxin producing species such as Fusarium sporotrichioides and Fusarium langsethiae could be identified with the designed molecular protocol designed based on Tri16 gene. In addition, weakly T-2 producing species can be identified with this method.

Keywords

Journal of Military Medicine
Volume 14, Issue 1 - Serial Number 47
March and April 2012
Pages 49-55

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