Abstract:MicroRNAs (miRNAs) are small endogenous non-coding RNAs with about 22 nucleotides emerging as a key regulator of gene activity at post-transcriptional level in eukaryotic cells. They negatively target mRNA that containing highly complementary sequences by either cleavage or translational repression. In this study, we analyzed the miR396 gene family in miRBase by utilizing bioinformatics methods. The results showed that miR396 was quite conserved in plant; it originated from ancient ancestor and experienced significant complex evolution; target prediction indicated miR396 mainly regulated GRF. Promoter analysis identified many stress-related cis-acting elements in the upstream region of miR396 genes, such as light, heat, anoxic, hormone, drought and biotic responses, which can bind transcription factors and participate in the regulatory activities of miR396 during stress. Our survey established foundation for deeply elucidate the regulatory mechanism of miRNA.