Corn stalks are a major source of agricultural waste in China that have the potential for more efficient utilisation. In this study, we designed substrate formulas with different proportions of corn stalks to cultivate Pholiota microspora. The substrate formula for P. microspora cultivation that could partially or completely replace sawdust with corn stalks was selected through the analysis of mycelial growth rates, fruiting body traits, yield, biological efficiency, nutrients, and mineral composition. Our results showed that the substrate formula T2 (38% wood chips and 38% corn stalks) resulted in the highest yield of 275.66 ± 2.87 g per bag, which was 6.60% higher than that of formula CK, and the highest biological efficiency of 90.75 ± 0.04%, which was 4.58% higher than that of CK, with no significant differences from CK in terms of fruiting body traits, nutrients, or mineral composition. The substrate formula T1 (19% corn stalks) led to mushroom yields with the highest mineral and amino acid contents and was thus more suitable for the cultivation of medicinal P. microspora. Therefore, substrates comprising a mixture of corn stalks and sawdust can be used as a novel, inexpensive, and high-yield alternative for the cultivation of P. microspora.
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