The release of trace metals caused by industrial effluents and anthropogenic activities has been recorded in the Xixi River estuary, southern China. However, a thorough understanding of the behavior of trace heavy metals in Xixi River sediments is lacking. A total of 12 sediment cores were collected in June and December in the upper estuary section and mouth of the estuary. Here, an in situ high-resolution sampling technique, namely, diffusive gradients in thin films (DGT), was employed to acquire profiles of trace element concentrations and the release of bioavailable metals from sediments in different seasons. A three-step Community Bureau of Reference (BCR) sequential extraction method was used to explore the chemical speciation of trace metals in different seasons and to thereby assess the release potential of trace elements in sediments. The BCR sequential extraction results showed that the trace metals Fe, Mn, Co and Pb were mainly in the residual fraction, which rarely influences living organisms. The total mobile fractions (F1 + F2 + F3) of all trace metals were higher in winter than in summer, suggesting that accumulation occurred from summer to winter. DGT measurements showed that the intensity of sulfate reduction was higher in summer than in winter because of the high temperatures and high organic matter in summer. The intensity of sulfate and Mn(III/IV) reduction increased from the upper estuary section to the lower estuary. Fe(III) reduction decreased in summer but increased slowly in winter. The Pearson correlation results showed that the release of DGT-labile Co in pore water was related to Mn(III/IV) reduction, while the release of DGT-labile Pb was basically not controlled by the Fe-Mn-S redox transition. Abnormally high DGT-labile Pb concentrations were observed at the sampling station (XR3) closest to the estuary in winter, which might have been caused by the high Pb content in the local micro-sediments.