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Applied and Environmental Microbiology, July 2001, p. 2966-2972, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.2966-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of Deoxynivalenol- and Nivalenol-Producing Chemotypes of Gibberella zeae by Using PCR

Theresa Lee,1 Dae-Woong Oh,1 Hye-Seon Kim,1 Jungkwan Lee,1 Yong-Ho Kim,2 Sung-Hwan Yun,2 and Yin-Won Lee1,*

School of Agricultural Biotechnology, Seoul National University, Suwon 441-744,1 and Division of Life Sciences, Soonchunhyang University, Asan 336-745,2 Korea

Received 22 January 2001/Accepted 12 April 2001

Gibberella zeae, a major cause of cereal scab, may be divided into two chemotypes based on production of the trichothecenes deoxynivalenol (DON) and nivalenol (NIV). We cloned and sequenced the gene cluster for trichothecene biosynthesis from each chemotype. G. zeae H-11 is a DON producer isolated from corn, and G. zeae 88-1 is a NIV producer from barley. We sequenced a 23-kb gene cluster from H-11 and a 26-kb cluster from 88-1, along with the unlinked Tri101 genes. Each gene cluster contained 10 Tri gene homologues in the same order and transcriptional directions as those of Fusarium sporotrichioides. Between H-11 and 88-1 all of the Tri homologues except Tri7 were conserved, with identities ranging from 88 to 98% and 82 to 99% at the nucleotide and amino acid levels, respectively. The Tri7 sequences were only 80% identical at the nucleotide level. We aligned the Tri7 genes and found that the Tri7 open reading frame of H-11 carried several mutations and an insertion containing 10 copies of an 11-bp tandem repeat. The Tri7 gene from 88-1 carried neither the repeat nor the mutations. We assayed 100 G. zeae isolates of both chemotypes by PCR amplification with a primer pair derived from the Tri7 gene and could differentiate the chemotypes by polyacrylamide gel electrophoresis. The PCR-based method developed in this study should provide a simple and reliable diagnostic tool for differentiating the two chemotypes of G. zeae.

* Corresponding author. Mailing address: School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Korea. Phone: 82-31-290-2443. Fax: 82-31-294-5881. E-mail: lee2443{at}snu.ac.kr.

Applied and Environmental Microbiology, July 2001, p. 2966-2972, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.2966-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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