The Zhengguang gold deposit is located in the southeast segment of the Duobaoshan polymetallic metallogenic belt, which is one of the largest ore deposit clusters in Heilongjiang Province, NE China and situated in the east segment of the Xing'an-Mongolia fold belt. In this study, the molybdenite Re-Os isotopic dating for the Zhengguang gold district yielded weighted mean age of 480±3 Ma, and the zircon U-Pb dating on the diorite-porphyrite and diorite yielded 206Pb/238U weighted mean ages of 478.3±3.7 Ma and 150.67±0.77 Ma, respectively. Accordingly, we infer that the early mineralization in the Zhengguang gold deposit occurred in Ordovician and was dominantly epithermal mineralization in the peripheral areas of the Duobaoshan porphyry metallogenic system related to diorite-porphyrite magmatic activities, and that the late mineralization in the deposit experienced metallogenesis of the Late Jurassic diorite magmatic activities. Therefore, the Zhengguang gold deposit is the product of multi-stage metallogenesis related to diorite-porphyrite and diorite. In addition, based on higher content of Re (295.9×10-6–394.7×10-6) in the molybdenite measured, we predict that the ore-forming materials were dominantly derived from the mantle, and contained some crustal materials.