The Shitouzhai Permian paleo-oil reservoir in Guizhou province is a typical example of numerous Upper Paleozoic reef-type paleo-oil reservoirs in the Yunnan, Guizhou and Guangxi regions. The homogenization temperatures of saline aqueous inclusions coexisting with the hydrocarbon inclusions in the three stage calcites are 77~lue="84" UnitName="℃">84 ℃, 91~lue="103" UnitName="℃">103 ℃ and 103~lue="155" UnitName="℃">155 ℃, respectively. It is suggested that the hydrocarbon charge of the Shitouzhai reservoir began during the middle Indosinian and reached to a peak during the late Indosinian to the early Yanshannian. The destruction of the reservoir occurred from the middle to late Yanshannian.