The speciation analysis of heavy metal pollutants is very important because different species induce different toxicological effects. Nanomaterial-assisted optical sensors have achieved rapid developments, displaying wide applications to heavy metal ions but few to metal speciation analysis. In this work, a novel colorimetric nanosensor strategy for mercury speciation was proposed for the first time, based on the analyte-induced aggregation of gold nanoparticles (Au NPs) with the assistance of a thiol-containing ligand of diethyldithiocarbamate (DDTC). Upon the addition of mercury species, because Hg-DDTC was more stable than Cu-DDTC, a placedisplacement between Hg species and Cu 2+ would occur, and thereby the functionalized Au NPs would aggregate, resulting in a color change. Moreover, by virtue of the masking effect of ethylenediaminetetraacetic acid (EDTA), the nanosensor could readily discriminate organic mercury and inorganic mercury (Hg 2+ ), and it is thus anticipated to shed some light on the colorimetric sensing of organic mercury. So, a direct, simple colorimetric assay for selective determination of Hg species was obtained, presenting high detectability, such as up to 10 nM for Hg 2+ and 15 nM for methylmercury. Meanwhile, the strategy offered excellent selectivity toward mercury species against other metal ions. The simple, rapid, and sensitive label-free colorimetric sensor for the determination of Hg species provided an attractive alternative to conventional methods, which usually involve sophisticated instruments, complicated processes, and long periods of time. More importantly, by using mercury as a model, an excellent nanomaterial-based optical sensing platform can be developed for speciation analysis of trace heavy metals, which can lead to nanomaterials stability change through smart functionalization and reasonable interactions.