Molecular cloning of a widely distributed microsatellite core sequence from the cultivated mushroom Agaricus bisporus

An Agaricus bisporus microsatellite with the tetranucleotide motif TATG tandemly repeated was isolated from an A. bisporus library enriched in repeated sequences. The use of the 16-mer oligonucleotide (TATG)4 indicates that many loci contain nearby copies of the microsatellite in opposite orientations. The wide distribution of the microsatellite in the A. bisporus genome was assessed (i) by polyacrylamide gel electrophoresis of the products generated by directed amplification of microsatellite-region DNA (DAMD) and (ii) by hybridization of these products with A. bisporus chromosomes separated by pulsed-field gel electrophoresis. This is, to our knowledge, the first microsatellite reported in the cultivated edible mushrooms. DAMD-PCR products were generated using DNA of three Pleurotus species (P. pulmonarius, P. sajor-caju, and P. florida), indicating that (TATG)4 repeats are also present in these cultivated species. The variability found within closely related strains indicates that such microsatellites are useful in fingerprinting and studying genetic variability in wild and commercial mushrooms.