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PCR amplification of the mating-type idiomorphs in Cryphonectria parasitica

Authors

  • R. E. Marra,

    Corresponding author
    1. Department of Plant Pathology, Cornell University, Ithaca, NY 14853, USA
      R. E. Marra. Fax: + 1520-6219190; E-mail:marra@u.arizona.edu
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      Present address: Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ B5721, USA.

  • M. G. Milgroom

    1. Department of Plant Pathology, Cornell University, Ithaca, NY 14853, USA
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R. E. Marra. Fax: + 1520-6219190; E-mail:marra@u.arizona.edu

Abstract

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.

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