[Objective] Silver nanoclusters were prepared using Pichia guilliermondii ZJC-1 for the detection of trace Cr(VI). [Methods] Fluorescent silver nanoclusters were synthesized using Pichia guilliermondii ZJC-1 that was acclimated to silver. The structures and fluorescence properties of silver nanoclusters were characterized. Then we explored the selective quenching effect of Cr(VI) on the silver nanoclusters fluorescence. We also established a linear relationship between silver nanoclusters fluorescence intensity and the concentration of Cr(VI). The effect of pH and other metal ions on the detection of Cr(VI) was also examined. [Results] There was a good linear relationship between Cr(VI) and the fluorescence intensity of silver nanoclusters (R²=0.9821). The detection limit was 184 μmol/L and the signal to noise ratio (SNR) was 3. Using this method to detect Cr(VI) in actual water samples (Songhua River, Majiagou River) the recovery rate was between 97.73% and 102.88%. [Conclusion] Silver nanoclusters with good fluorescence properties can detect trace concentrations of Cr(VI) simply and rapidly.