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Verticillium dahliae  is a cosmopolitan, soilborne fungus that causes a significant wilt disease on a wide variety of plant hosts including economically important crops, ornamentals, and timber species. Clonal expansion through asexual reproduction plays a vital role in recurring plant epidemics caused by this pathogen. The recent discovery of recombination between clonal lineages and preliminary investigations of the meiotic gene inventory of  V. dahliae  suggest that cryptic sex appears to be rare in this species. Here we expanded on previous findings on the sexual nature of  V. dahliae . Only 1% of isolates in a global collection of 1120 phytopathogenic  V. dahliae  isolates contained the  MAT1-1  idiomorph, whereas 99% contained  MAT1-2 . Nine unique multilocus microsatellite types comprised isolates of both mating types, eight of which were collected from the same substrate at the same time. Orthologs of 88 previously characterized sex-related genes from fungal model systems in the Ascoymycota were identified in the genome of  V .  dahliae , out of 93 genes investigated. Results of RT-PCR experiments using both mating types revealed that 10 arbitrarily chosen sex-related genes, including  MAT1-1-1  and  MAT1-2-1 , were constitutively expressed in  V .  dahliae  cultures grown under laboratory conditions. Ratios of non-synonymous (amino-acid altering) to synonymous (silent) substitutions in  V .  dahliae MAT1-1-1  and  MAT1-2-1  sequences were indistinguishable from the ratios observed in the  MAT  genes of sexual fungi in the  Pezizomycotina . Patterns consistent with strong purifying selection were also observed in 18 other arbitrarily chosen  V .  dahliae  sex-related genes, relative to the patterns in orthologs from fungi with known sexual stages. This study builds upon recent findings from other laboratories and mounts further evidence for an ancestral or cryptic sexual stage in  V. dahliae .

Maintenance of Sex-Related Genes and the Co-Occurrence of Both Mating Types in Verticillium dahliae