The cDNA and promoter sequence of FnMYB24 gene were cloned from Fragaria nilgerrensis Schltdl. by RTPCR. Bioinformatics analysis showed that the cDNA sequence of FnMYB24 was 1 033 bp (GenBank accession number MN879283) length, and its open reading frame (ORF) was 609 bp length, encoding 202 amino acids, including a conserved MYB_DNAbinding domain. Homogeneous analysis showed that the amino acid sequence encoded by FnMYB24 gene was 92.75% similar to that of encoded by Fragaria vesca. The promoter sequence of 718 bp upstream of the transcription initiation site (GenBank accession number MN879285) was further cloned, which was predicted to contain several cisacting elements such as hormone response elements and optical regulation elements. By constructing pFnMYB24∷GUS expression vector and carrying out instantaneous tobacco transformation, it was found that the pFnMYB24 promoter had transcriptional activity and could drive the expression of FnMYB24 gene. Realtime quantitative PCR results showed that the expression of MYB24 gene in the leaves of resistant variety F. nilgerrensis Schltdl. and susceptible cultivar ‘Miaoxiang No.3’ was upregulated after inoculation with Colletotrichum gloeosporioides, but the expression level of MYB24 in ‘Miaoxiang No.3’ was always lower than that in F. nilgerrensis Schltdl. This gene was also induced by SA since the MYB24 expression levels in two strawberry varieties after SA treatment were higher than those of controls. Our results suggest that MYB24 may involved in regulate resistance to strawberry anthracnose, this study laid foundation for further investigate the function of MYB24 confers resistance to strawberry anthracnose.