A simple genetic method for identification of mycotoxigenic fungi-Development of heteroduplex panel analysis and its field application-

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  • 分子生物学的手法を用いたカビの簡便な同定法
  • - Development of heteroduplex panel analysis and its field application -

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Abstract

A novel method for heteroduplex panel analysis (HPA) utilizing fragments of the PCR amplified ITS regions of rDNA was developed. The method involves formation of heteroduplexes with a set of reference fragments amplified from A. flavus, A. parasiticus, A. tamarii<i> and <i>A. nomius, and subsequent minislab gel electrophoresis. The test panel is compared with species-specific standard panels (F-1, P-1, T-1 and N-1), generated by pairwise reannealing among four reference fragments. Application of HPA to a field study demonstrated that a new genotype of FP-1 has been predominantly distributed throughout sugarcane field soil in the southernmost islands of Japan. The high prevalence of type FP-1 may reflect its adaptation to the soil environment. The type FP-1 isolates were also isolated from the sugarcane fields in Vietnam, but not find them in our culture collection derived from other sources. CBS 108.30 originated from sugarcane in Egypt was the only one strain identified as type FP-1. These results imply that type FP-1 may be associated with sugarcane fields in Asia. Type FP-1 is consisted of the morphologically intergrading isolates between A. parasiticus and A. flavus. The phylogenetic analysis based on the ITS sequences indicated that type FP-1 formed an independent clade positioned between A. parasiticus and A. flavus, and was more closely related to the former species. HPA also demonstrated that A. nomius has been distributed in sugarcane field soil and sugarcane stems in Japan and Vietnam. All of the tested isolates of A. nomius and type FP-1 were able to produce aflatoxins B and G. These fungi are probably responsible for aflatoxin contamination of crude sugar produced from sugarcane.

Journal

  • JSM Mycotoxins

    JSM Mycotoxins 58 (1), 29-40, 2008

    Japanese Society of Mycotoxicology

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