Pleurotus species are edible mushrooms that possess the ability to degrade lignocellulose and are cultivated on lignocellulosic substrates to produce fruiting bodies that are appreciated for their nutritional and culinary values. Radial growth rate (ur), mycelial biomass, and intracellular glycogen and protein contents were evaluated in the colonies of two strains of P. ostreatus (strains 26 and 50) and one strain of P. pulmonarius (strain 35) grown on barley straw extract agar, corn stover extract agar (CSEA), wheat straw extract agar, and glucose-yeast extract agar (GYEA). The nutritional composition of the Pleurotus fruit bodies cultivated on barley straw, corn stover, and wheat straw were also determined. Of all of the strains, P. ostreatus 50 showed the highest ur on the CSEA and GYEA, and P. ostreatus 26 showed the greatest mycelial biomass production on the lignocellulosic waste-based agar media (LWAM). All of the Pleurotus strains showed the greatest and lowest glycogen contents on the CSEA and GYEA, respectively. These results showed that the LWAM are more suitable for mycelial growth than a glucose-based agar medium, and that Pleurotus fruit bodies cultivated on corn stover had the highest nutritional value compared with fruit bodies grown on the wheat and barley substrates.