miRNAs are a class of non-coding and small RNAs (~ 21 nucleotides) performs a key function in plant growth and physiology by regulating gene expression in post-transcriptional level. Since there has not been a comprehensive study on identification of the miRNAs of safflower (Carthamus tinctorius L) using in silico method, in current study, 124 potential miRNAs belonging to 26 conserved families were identified and two miRNAs were validated using qRT-PCR technique. The 29 miRNA families were significantly different in size. Safflower pre- miRNAs greatly varied from 57 to 317 nt in length with an average of 94.56 ± 38.1 nt. Another result of this study was the success in determining miR169, miR395 and miR397 clusters in the safflower genome. In the present study, the family members of miRNAs including miR393, miR477, miR530, miR6111, miR6113, and miR6114were recognized which have not previously been reported. A total of 84 potential target genes were predicted for the identified safflower miRNAs including transcription factors, proteins involved in DNA replication, and metabolic enzymes. The results of this study showed the existence of conserved miRNAs in safflower and may be important role in response to environmental stress.