Abstract: ObjectiveThe important role of miR169(vvi-miR169)gene family in Vitis vinifera during the growth and stress response was clarified,which could provide reference for its application in molecular breeding and resistance breeding in Vitis vinifera.MethodBy utilizing miRNA,Phytozome and NCBI database,ClustalX 2.1,MEGA 6,RNAfold,WebServer, psRNATarget and other online softwares,bioinformatics analysis for vvi-miR169 gene family was conducted,including its se-quences,distribution characteristics,the secondary structure,phylogenetic tree and target gene regulation function.ResultTwenty-five vvi-miR169 homologous gene sequences which were searched in the miRBase distributed on six chromosomes. Chr11 had the most gene sequences(17 sequences)distributed on. There were two sequences on Chr01,Chr04 and Chr14, while only one on Chr08 and Chr17. Base of vvi-miR169 gene family mature sequence was of highly conservation. Its precur-sor sequence could form a stable two stem loop structure,and miR169 gene sequence on the same chromosome showed close genetic relationship. It was predicted that vvi-miR169 gene family contained 21 target genes,most miRNA genes had multiple target genes,different miRNA gene shared the same targets,and 22 miRNA genes contained the same target gene of GS-VIVT01015120001(transcription factor NF-YA).Conclusionvvi-miR169 gene family sequences are highly conserved and characterized by obvious family features with similar regulatory function. Transcription factor NF-YA is the main target gene of vvi-miR169 gene family,and is also the main stress-resistance regulation element in the growth and development of V. vinifera.