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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.

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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