Ganoderma lingzhi is a kind of precious medicinal mushroom. MicroRNAs (miRNAs) play an important role in the regulation of a broad range of biological processes. In this study, we first report on the miRNAs of the G. lingzhi sporocarp. In total, 132 known miRNAs and 34 putative candidate (PC) miRNAs were identified for the first time in G. lingzhi sporocarp. Through target gene prediction, 111 of 132 known miRNAs had targeted genes, 58 of them had more than one, and most predicted targeted genes might be regulated by more than one miRNA; functional enrichment by KEGG analysis demonstrated that target genes related to cancer and signal transduction occupied the largest proportion. A total of seven miRNAs (four known miRNAs and three PC miRNAs) that had high abundance were selected for expression confirmation by quantitative real-time PCR (qRT-PCR) and northern blotting. This is the first comprehensive study of the miRNA composition in G. lingzhi, which resulted in the addition of a significant number of novel miRNA sequences to miRBase and laid the foundation for further understanding of the function of miRNAs in the regulation of G. lingzhi. (c) 2016 Published by Elsevier B.V. on behalf of The Mycological Society of Japan.