[Aim] Insects show different biological characteristics as they develop under variable environmental conditions, and their epigenetic regulatory mechanisms are worthy of study. The longhorn beetle, Monochamus alternatus is a vector of the pinewood nematode, the causal agent of the devastating pine wilt disease. Numerous studies show differences in morphology and development rate of M. alternatus larvae between laboratory and field populations, yet their epigenetic response mechanism is not clear. Therefore, this paper aims to describe the difference between these two populations by microRNA expression profiling, in order to provide a reference for epigenetic research in M. alternatus.[Method]High-throughput sequencing of microRNA using the illuminaHiSeq 2000 platform resulted in the construction of microRNA libraries from two laboratory-bred and two filed-collected beetle populations. The identification of conserved microRNAs and prediction of new microRNAs was also done. In addition, differential expression of microRNA was analyzed, and the function of target genes for these miRNA was predicted by GO annotation and KEGG pathway enrichment analysis.[Result] A total of 16 and 14 microRNA were identified in the epidermis and midgut of indoor-reared larvae, respectively, while 13 microRNA each were identified in the epidermis and midgut of the field-collected larvae. Compared to the epidermis, miRNA expression in the midgut was higher. Compared to the wild population, the expression of microRNA in the M. alternatus in the laboratory-reared colony was higher. Here, 17 microRNA expression levels changed significantly, such as novel-mir-62127, novel-mir-184731, novel-mir-290819 were significantly up-regulated, and novel-mir-251851 were significantly down-regulated. The function of the target gene of the differentially expressed miRNA mainly points to sugar metabolism such as amino sugar metabolism, chitin metabolism, and lipid metabolism, mainly those of glycerophospholipid and fatty acids.[Conclusion] There were obvious differences in the microRNA inventory between field-collected and laboratory-bred mature larvae of M. alternatus, and the microRNA expression profiles of different tissues were also significantly different. It is suggested that M. alternatus under labratory-reared conditions has the characteristics of epigenetic regulation. This lays the foundation for further studying the microRNA regulation mechanism not only for M. alternatus' development and metabolism, but may also uncover similar patterns of epigenetic differences between wild and laboratory populations for other insects reared for research purposes.