Seed germination of spinach (Spinacia oleracea L.) is partially inhibited by a high germination temperature (35 °C). Tolerance of high germination temperatures varies widely depending on the variety used. We ascertained that seed germination of these spinach varieties was thermoinhibited at 35 °C and secondary dormancy was not induced as seeds germinated when transferred to optimum germination conditions (20 °C). Treatment with 99% oxygen and 10 ppm kinetin significantly increased germination of thermoinhibited varieties at 35 °C. During heat stress, all organisms produce heat shock proteins (HSPs), which may function as molecular chaperons, are possibly required for the development of thermotolerance, and may be crucial for cell survival during heat stress. Western blotting of SDS-PAGE gels using antibodies to various heat shock proteins indicated that spinach varieties with the highest degree of thermotolerance have higher levels of HSP expression than varieties with the lowest degree of thermotolerance during germination. These results suggest that thermotolerance could be further improved, either through a breeding program or possibly by genetic engineering.