PCR amplification of the mating-type idiomorphs in Cryphonectria parasitica.
In the laboratory, the ascomycete fungus Cryphonectria parasitica is rarely self-fertile, and has a self-incompatibility system that resolves into two intersterility groups, controlled by a single locus. In natural populations, however, self-fertilization occurs frequently. In this report, we show that the C. parasitica self-incompatibility locus (MAT) comprises two idiomorphs (alleles that are highly divergent in sequence), conforming to the paradigm of self-incompatibility as described for other ascomycetes. Starting with a fragment putatively from the MAT-2 idiomorph, we used a PCR-based cloning approach to identify 3.5- and 2-kb sequences unique to MAT-1 and MAT-2 isolates, respectively. These sequences were then used to design idiomorph-specific PCR primer pairs, allowing us to efficiently identify the mating types of isolates, a crucial component of our research on the environmental and genetic factors underlying this mixed mating system.
Marra
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Department of Plant Pathology, Cornell University, Ithaca, NY 14853, USA. marra@u.arizona.edu
Milgroom
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D000483:Alleles; D001203:Ascomycota / Q000235:genetics* / Q000502:physiology; D005800:Genes, Fungal / Q000235:genetics*; D049770:Genes, Mating Type, Fungal; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA
D006609:High Mobility Group Proteins