Rapid and effective identification of A mating-type loci in Volvariella volvacea by polymerase chain reaction-restriction fragment length polymorphism

Abstract

Mating-type genes determining the compatibility of monosporous are considered as the most important markers in breeding. Prior to mating tests, identification of compatible strains can improve the breeding program. The mating type discrimination and mating were determined by A mating-type loci in Volvariella volvacea, and thus it was important for us to investigate V. volvacea A mating-type genes. The hyperdiversity of DNA sequences at A mating-type loci, however, has made isolating of these genes very difficult. In this study, Polymerase chain reaction-restriction fragment length polymorphism method was developed for rapid and effective amplification and identification of A mating-type genes from V. volvacea. A total of eight different A mating-type loci were distinguished according to the restriction enzymes pattern analysis. The alignment of DNA sequence and phylogenetic analysis supports that it was feasible to identify A mating-type loci using polymerase chain reaction-restriction fragment length polymorphism analysis. This approach allowed differentiation of A mating-type genes within one day and did not use hybridization steps.