In the process of diagenesis and burial of sedimentary basins, basin fluid activities participate in the process of hydrocarbon accumulation and metal mineralization. Understanding the evolution of basin fluid is of great significance in revealing the related hydrocarbon accumulation and mineralization. Paleo-reservoirs are closely associated with Carlin-type gold deposits in the Nanpanjiang-Youjiang Basin, South China. Calcite, the fluid activity product, is closely related to bitumen and gold-bearing pyrite. By integrating petrographic, cathode luminescence, and fluid inclusion analysis, as well as the relevant chronological results of predecessors, this paper attempts to establish the relationship between fluid evolution, hydrocarbon accumulation, and gold mineralization. Two types of calcite (black/gray and white) developed in the Banqi-Yata-Laizishan area, the Nanpanjiang-Youjiang Basin. Black/gray calcite is symbiotic with bitumen and features dark red colors in cathode luminescence. Many hydrocarbon inclusions developed along with fluid inclusion analysis at low homogenization temperatures (65.7~173.1 °C). Combining the previously reported U-Pb ages (~250−230 Ma) of this kind of calcite with some geochemistry data on the associated reservoir and gold deposit, this calcite records the consecutive hydrocarbon accumulation and Carlin-type gold mineralization from the Late Permian to the Late Triassic periods controlled by Indosinian tectonic movement. The white calcite featuring bright red in cathodoluminescence is symbiotic with gold-bearing pyrite and realgar, and the associated fluid inclusions have high homogenization temperatures (128.2~299.9 °C). Combined with regional tectonic background and isotopic chronology (~140−106 Ma), it seems to record the early Cretaceous Carlin-type gold mineralization controlled by the subduction of the paleo-Pacific plate in the late Yanshanian period.