Single spore derived Pleurotus spp. isolates from four commercial lines (two P. sapidus, one P. florida, and one P. ostreatus) and from two interspecific hybrids (P. sajor-caju × P. sapidus) were analyzed for single locus and joint segregation of 25 allozyme encoding loci. The two alleles at the individual loci departed significantly in their segregation from a 1:1 Mendelian ratio in 26% of the intraspecific and 29% of the interspecific tests. Six linkage groups were identified as follows: Dia-1 ~ Est-5; Tpi ~ Pgd-2 ~ Skdh; (Fum) ~ Pgm-2 ~ Pgd-1 ~ PepLgg-1 ~ Gr-2; Ndh ~ Gr-1; Np ~ PepGl-1 ~ Aat-2 ~ Pgk ~ Mup; and Gr-4 ~ Mdh-1. The duplicate loci coding for GR, PEP-LGG, PGM, and PGD were both not linked to each other and not part of duplicate linkage groups. Six loci were not shown to be linked to any other loci (Lap, Pgm-1, Ha, Gpi, PepPap, and PepLgg-2), although the latter two loci were only tested against four and five loci, respectively. The first linkage map of 19 allozyme encoding loci for the Pleurotus genome is presented.Key words: Pleurotus, allozymes, linkage map, inheritance, edible fungi.