Development of STS markers suitable for marker-assisted selection of sporeless trait in oyster mushroom, Pleurotus pulmonarius

In the cultivation of edible mushrooms including Pleurotus pulmonarius, the enormous number of spores produced by fruiting bodies adversely affects the health of mushroom farmers, mushroom cultivation facilities and the genetic diversity of natural populations. A sporeless mutant of P. pulmonarius (TMIC-30058) has been introduced with a high potential for breeding sporeless cultivars to reduce these adverse effects. We report here the conversion of the amplified fragment length polymorphism (AFLP) marker, closely linked to a sporeless locus, to a sequence tagged site (STS) marker for use in marker-assisted selection (MAS). By mainly bulked segregant analysis-based AFLP (BSA-AFLP), a total of 18 markers were identified the linkage to the sporeless locus with genetic distances ranging from 0 (co-segregation) from 3.4 cM. Out of them, 2 AFLP markers could be converted into STS markers, SD192 and SD296, and their implementations in various genetic backgrounds were validated by the amplification test against 15 wild type isolates from different geographical origin. Furthermore, we could amplify these 2 STS markers’ fragments by using multiplex PCR reproducibly. These results indicated that 2 STS markers can be used in MAS targeted at the sporeless trait in P. pulmonarius breeding.