In this study, we cloned a heat shock transcription factors (HSFs) gene, ZmHSF05 from maize (Zea mays) seedlings by RTPCR amplification and sequencing. The open reading frame of ZmHSF05 is 1 080 bp in length, which encodes 359 amino acid residues. Sequence analysis indicated that ZmHSF05 possessed a conserved DNA binding domain (BD), an activator domain (AD/AHA) and a nuclear localization signal (NLS) site. Subcellular localization analysis showed that pEGADZmHSF05 fusion protein only distributed in the nucleus, indicating that ZmHSF05 gene localized in the nucleus. Cisacting element analysis of the 2 102 bp sequence upstream of the ATG of ZmHSF05 gene initiation codon revealed that the promoter region of ZmHSF05 contained several hormone and stress response related ciselements. qRTPCR result indicated that ZmHSF05 was highly expressed in root, and responsive to heat, drought and salt stresses. ZmHSF05 overexpress transgenic lines of Arabidopsis exhibited more thermotolerant phenotype under heat stress, including higher survival rate and cell membrane thermostability than wild type plants. These results indicated ZmHSF05 could be a positive regulator in heat stress response pathway.