Abstract: Based on the transcriptome database of Dendrocalamus farinosus,a MYB transcription factor named DfMYB3 was cloned from the leave of D. farinosus. Its open reading frame length was 1 287 bp,encoding 428 amino acids,and GenBank accession Number was KY963358. The conserved domain analysis showed that DfMYB3 had the typical SANT domains and DNA-Binding domains. Phylogenetic tree analysis showed that DfMYB3 clustered with R2 R3-MYB transcription factors in Saccharum,Arabidopsis and Populus. Subcellular localization showed that DfMYB3 protein was expressed in both the nucleus and cell membrane,and was more significant in the nucleus. The sequence of 5′flanking of DfMYB3 gene about 2 000 bp was cloned by chromosome walking method. Analysis of PlantCARE online software showed that the sequence had typical promoter characteristics,and contained hormones responsive elements such as GA,ABA,MeJA as well as stress responsive elements such as drought. The expression level of DfMYB3 gene was significantly up-regulated after 100 μmol·L-1 GA and 100 μmol·L-1 ABA treatment,indicating that DfMYB3 gene was involved in response to GA and ABA treatments. In order to explore the function of DfMYB3 promoter,the expression vector of GUS gene fused with DfMYB3 promoter was constructed and genetically transformed into tobacco. The results showed that GUS signal was detected in both leaf and stem of transgenic tobacco,especially in the veins.