Abstract: Heat stress restricts grape production, especially in facility cultivation. Analysis of SSR markers and classification of HSFs which are major regulators in heat tolerance could be important in grape heat tolerance breeding. In this study, 19 HSF sequences of Vitis vinifera were downloaded from Phytozome. Based on these sequences of VvHSFs, their lengths, molecular weights and isoelectric points were predicted. Furthermore, multiple alignment and phylogenetic analysis of these 19 HSFs were carried out with AtHSFs as reference sequences. 10 HSFAs, 7 HSFBs and 2 HSFCs were found in grape genome. According to SSR sites in VvHSF, the specific primers were designed for the PCR using‘Cabernet Sauvignon’as template. These data showed that grape genome harbored 3 classes of HSFs containing SSR sites which could be used for designing molecular markers. This work would be a basis for molecular marker assistant breeding in grape thermotolerance.