In the present study, potentially edible and nutritious Pleurotus spp. i.e., P. cystidiosus, P. floridanus, P. ostreatus were collected to figure out their cultural characteristics and to find out the optimum requirements for their mycelial growth which are the pre-requisite requirements for their successful cultivation. Their mycelial features were observed on the different nutrient media at different temperatures. Mycelium of each species was assessed morpho-anatomically. Mycelium growth pattern and texture of three Pleurotus species were irregular and cottony while a special anamorphic stage (Antromycopsis macrocarpa) in the form of Coremium and coremioliquid appeared during the cultural growth of P. cystidiosus. Anatomically mycelium of each species was septate and clampedwith the addition of toxocyst and arthroconidia production in the anamorphically grown myceliumof P. cystidiosus, which is unreported feature for this species from Pakistan. P. floridanus took more time to completely colonize the petriplates on all media followed by P. cystidiosus and P. ostreatus. All species have spawn production potential but spawn production rate was more of P. ostreatus followed by P. floridanus and P. cystidiosus. The linear growth rate was assessed on the mixed substrate of sawdust and wheatstraw. P. floridanus, P. ostreatus, P. cystidiosus mycelium showed their best colonization rate at 16ᵒC, 22ᵒC, and 28ᵒC respectively. From these results it could be assessed that these nutritionally and medicinally significant Pleurotus spp. possess the cultivation potential and their cultivation on large scale can meet the nutritious need of the growingpopulation of the world.