Little is known about mercury (Hg) biomagnification in the subtropics, aquatic systems with high species diversity resulting in complex food webs. High atmospheric Hg emissions and ubiquitous reservoir fisheries may lead to elevated Hg bioaccumulation in Chinese freshwater fishes. However, stocking practices using fast-growing species can result in low fish total Hg (THg) concentrations. Here, we describe Hg transfer within the fish food web of a large subtropical reservoir, Qiandao Hu (Xin'anjiang reservoir) situated in eastern China. We measured food web Hg biomagnification and THg concentrations in 33 species of stocked and wild fishes. Mercury concentrations in most fishes were low, though we also found high Hg concentrations in wild top predators. The food web structure, assessed using stable isotopes of carbon (δ(13)C) and nitrogen (δ(15)N), demonstrated a high degree of omnivory and a long food chain. THg concentrations were highly correlated with fish δ(15)N values. The regression of log10THg against δ(15)N revealed the overall Hg biomagnification rate was low. This study shows that where long food chains exist in subtropical reservoirs, elevated Hg accumulation in top predators can occur despite a low Hg biomagnification rate.