Seven precursors (premiRNAs) and seven mature sequence of the miR171 family were obtained for analyzing the evolutionary characterization and its expression from Dimocarpus longan Lour. (longan) mRNA and miRNA databases. The prediction results using mfold showed that the sequence of premiR171 family members could form a typical secondary structure,and the minimum folding free energy (dG) in the range of -39.37 to -55.13 kal/mol. Other members of the mature sequence matched exactly with vvimiR171a, ptcmiR171f and ctrmiR171 except premiR171scaffold112 and premiR171fscaffold38. NJ phylogenetic tree analysis showed higher sequence homology with that from Citrus reticulata and Populus trichocarpa. Target prediction showed that the main target of longan miR171 family were Scarecrowlike6,which was the same as miR171 target of other plants. qPCR results indicated that premiR171 family were strong expressed in the male flower and low in the fruit. It was suggested that the miR171 family was highly conserved during the evolution and might play important roles in the development of male flowers in longan.