Intra-specific and inter-specific conservation of mating-type genes from the discomycete plant-pathogenic fungi Pyrenopeziza brassicae and Tapesia yallundae.

ABSTRACT

In previous work, four genes involved in mating-type determination were cloned from reference strains of Pyrenopeziza brassicae; three genes, PAD1, PMT1, and PHB1 (re-named henceforth as MAT-1-1, MAT-1-4, and MAT1-3, respectively), are encoded by the MAT-1 idiomorph, and one gene, PHB2 (re-named MAT-2), by the corresponding MAT-2 idiomorph. To assess MAT gene organisation within field-populations of P. brassicae, 30 field-isolates of both mating-types from different geographical locations were analysed by PCR using primers designed for the MAT genes of P. brassicae. The results indicate that mating-type gene structure and organisation within these isolates is conserved and is consistent with the mating-type designations established by crossing experiments. The four P. brassicae MAT genes were then used as probes against gel blots of the genomic DNA of a discomycete Tapesia yallundae from the same family (Dermateaceae, order Helotiales) and one, Ascobolus stercorarius, from a distantly related family (Ascobolaceae, order Pezizales), in order to determine whether P. brassicae MAT-gene homologs were present. MAT-specific hybridisation signals were obtained with T. yallundae using all four probes. In particular, MAT-1 DNA of T. yallundae gave a strongly hybridising signal with MAT-1-4 (PMT1), a putative metallothionein gene found in the P. brassicae MAT-1 idiomorph but not in any other MAT idiomorph examined to-date. No MAT-specific hybridisation was obtained with A. stercorarius. A fragment of the MAT-2 gene of T. yallundae was obtained by PCR using degenerate primers designed to amplify the high-mobility group (HMG) domain present in other ascomycete MAT genes. Sequencing of this PCR product revealed similarities to MAT HMG domains from other ascomycetes with the greatest degree of similarity exhibited with P. brassicae. The T. yallundae HMG-DNA sequence was shown to co-segregate with mating type (MAT-2) in progeny from a sexual cross.