Genetics, Vol. 163, 677-684, February 2003, Copyright © 2003

Trichothecene Nonproducer Gibberella Species Have Both Functional and Nonfunctional 3-O-Acetyltransferase Genes

Makoto Kimuraa,b, Takeshi Tokaia,c, Gentaro Matsumotoa,d, Makoto Fujimurac, Hiroshi Hamamotoe, Katsuyoshi Yoneyamad, Takehiko Shibatab, and Isamu Yamaguchia,e
a Laboratory for Remediation Research, Plant Science Center, RIKEN, Wako, Saitama 351-0198, Japan,
b Cellular and Molecular Biology Laboratory, RIKEN, Wako, Saitama 351-0198, Japan,
c Faculty of Life Science, Toyo University, Itakura, Gunma 374-0193, Japan,
d Faculty of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan
e Laboratory for Adaptation and Resistance, Plant Science Center, RIKEN, Yokohama, Kanagawa 230-0045, Japan

Corresponding author: Makoto Kimura, Plant Science Center, RIKEN, 2-1 Hirosawa, Saitama 351-0198, Japan., mkimura{at}postman.riken.go.jp (E-mail)

Communicating editor: M. S. SACHS

The trichothecene 3-O-acetyltransferase gene (FgTri101) required for trichothecene production by Fusarium graminearum is located between the phosphate permease gene (pho5) and the UTP-ammonia ligase gene (ura7). We have cloned and sequenced the pho5-to-ura7 regions from three trichothecene nonproducing Fusarium (i.e., F. oxysporum, F. moniliforme, and Fusarium species IFO 7772) that belong to the teleomorph genus Gibberella. BLASTX analysis of these sequences revealed portions of predicted polypeptides with high similarities to the TRI101 polypeptide. While FspTri101 (Fusarium species Tri101) coded for a functional 3-O-acetyltransferase, FoTri101 (F. oxysporum Tri101) and FmTri101 (F. moniliforme Tri101) were pseudogenes. Nevertheless, F. oxysporum and F. moniliforme were able to acetylate C-3 of trichothecenes, indicating that these nonproducers possess another as yet unidentified 3-O-acetyltransferase gene. By means of cDNA expression cloning using fission yeast, we isolated the responsible FoTri201 gene from F. oxysporum; on the basis of this sequence, FmTri201 has been cloned from F. moniliforme by PCR techniques. Both Tri201 showed only a limited level of nucleotide sequence similarity to FgTri101 and FspTri101. The existence of Tri101 in a trichothecene nonproducer suggests that this gene existed in the fungal genome before the divergence of producers from nonproducers in the evolution of Fusarium species.




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