Abstract: miRNA is a relatively special and highly conserved RNA, which plays an important role in gene regulation network. In this study, with the help of Illumia sequencing platform and the Solexa sequencing analysis of miRNA in Larix olgensis, we ultimately selected 15 294 797 clean reads, in which rRNA is 2 991 972 reads, tRNA 65 264 reads, snRNA 1 670 reads and snoRNA 467 reads, and ncRNAs and repeat sequences contain 6 421 reads. Un-annotated miRNA has 12 229 003 reads, accounting for 79.96% of the total RNA notes. The miRDeep2 was used to screen the miRNA, and 78 new miRNAs were obtained overall, which belong to the miR164, miR166, miR950, miR160, and miR396 families. Through further analysis of the target gene notes, we obtained 333 corresponding target genes, and their functions include transcription factors, unknown functional proteins, binding proteins, elongation factors, signal transduction, cell wall and cell membrane biosynthesis involved in regulation of plant growth and development process.