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High pressure thermal sterilization (HPTS) is a combination of high pressure (100-1000 Mpa) and heat treatment (50-80 ℃),an innovative technology to preserve food.This study aims to further explore the inactivation effect and mechanism of on Bacillus subtilis spores.Bacillus subtilis spores were cultivated on culture medium rich in Mn2+ at 37℃ for 7 days then suspended in sterile de-ionized water.By means of plate count the inactivation effect of HPTS on spore suspension was evaluated,spore membrane permeability was measured by flow cytometry staining;extracellular ultraviolet absorption substance content (A 260 and A 280) was used to characterize the core damage;flora germination rate and the 2,6-pyridine dicarboxylic (DPA) acid release rate measurement was to observe the spore germination.