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Applied and Environmental Microbiology, October 2000, p. 4378-4382, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

PCR-Based Identification of MAT-1 and MAT-2 in the Gibberella fujikuroi Species Complex†

Emma T. Steenkamp,1,* Brenda D. Wingfield,1 Teresa A. Coutinho,1 Kurt A. Zeller,2 Michael J. Wingfield,1 Walter F. O. Marasas,3 and John F. Leslie2

Tree Pathology Co-Operative Programme, Forestry and Agricultural Biotechnology Institute, Departments of Genetics, Microbiology, and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa1; Department of Plant Pathology, Throckmorton Plant Sciences Center, Kansas State University, Manhattan, Kansas 66506-55022; and Programme on Mycotoxicology and Experimental Carcinogenesis, Medical Research Council, Tygerberg 7505, South Africa3

Received 3 April 2000/Accepted 12 July 2000

All sexually fertile strains in the Gibberella fujikuroi species complex are heterothallic, with individual mating types conferred by the broadly conserved ascomycete idiomorphs MAT-1 and MAT-2. We sequenced both alleles from all eight mating populations, developed a multiplex PCR technique to distinguish these idiomorphs, and tested it with representative strains from all eight biological species and 22 additional species or phylogenetic lineages from this species complex. In most cases, either an ~800-bp fragment from MAT-2 or an ~200-bp fragment from MAT-1 is amplified. The amplified fragments cosegregate with mating type, as defined by sexual cross-fertility, in a cross of Fusarium moniliforme (Fusarium verticillioides). Neither of the primer pairs amplify fragments from Fusarium species such as Fusarium graminearum, Fusarium pseudograminearum, and Fusarium culmorum, which have, or are expected to have, Gibberella sexual stages but are thought to be relatively distant from the species in the G. fujikuroi species complex. Our results suggest that MAT allele sequences are useful indicators of phylogenetic relatedness in these and other Fusarium species.


* Corresponding author. Mailing address: FABI, University of Pretoria, Pretoria 0002, South Africa. Phone: 27-12-420-3948. Fax: 27-12-420-3946. E-mail: emma.steenkamp{at}fabi.up.ac.za.

dagger Contribution 00-322-J from the Kansas Agricultural Experiment Station, Manhattan.


Applied and Environmental Microbiology, October 2000, p. 4378-4382, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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