In order to make full use of the wild resources of Erianthus fulvus and mining valuable resistance genes to enrich the candidate gene library of transgenic sugarcane breeding, we cloned MYB gene from E. fulvus ‘991’ by combined with the transcriptome analysis and RTPCR technology. We also curried out bioinformatics analysis and stress expression analysis. This study provided theoretical foundation for further analysis cold tolerance mechanism of E. fulvus and transgenic sugarcane breeding. The results showed that: (1) E. fulvus MYB was successfully cloned and named EfMYB1(GeneBank ID: ON586646). (2) Bioinformatics analysis showed that the full length of EfMYB1 gene was 1 000 bp, had an ORF with 759 bp and encoded 251 amino acids. The EfMYB1 protein has a conserved SANT domain and multiple phosphorylation sites, and no transmembrane structure and signal peptide. The secondary structure and tertiary structure of EfMYB1 protein were mainly αhelix and random coil. EfMYB1 protein has the highest similarity and closest genetic distance with MYB of Miscanthus lutarioriparius. (3) The results of qRTPCR analysis showed that the relative expression level of EfMYB1 gene were gradually increased with continuous low temperature stress in root and leaf tissues of E. fulvus, and reached the largest level at 72 h, while almost unchanged in stem. In addition, EfMYB1 gene expression was induced by Methyl Jasmonate (MeJA) treatment, and reached the highest value at 6 h. But the expression level of this gene was significantly decreased under Abscisic acid (ABA) treatment. This study demonstrated that EfMYB1 gene belongs to low temperature stress response gene and may be involved in E. fulvus response low temperature stress.