To investigate the mechanism of cadmium tolerance in a cadmium-resistant Azomonas agilis PY101 that produces a specific fluorescent pigment promoted by cadmium, we carried out Tn5 mutagenesis and isolated four pigment-deficient mutants. In these mutants, Ppg1, Ppg2, and Ppg3 remarkably reduced the pigment production to 15.3%, 11.2%, and 13.9%, respectively. Especially, Ppg4 mutant did not produce the pigment at all. None of the mutants grew in the presence of 1500 ppm of CdCl2 in growth medium, and they exhibited differential sensitivities to cadmium. Ppg1, Ppg2, Ppg3, and Ppg4 mutants were sensitive to 900 ppm, 1100 ppm, 1000 ppm, and 800 ppm of CdCl2, respectively. These mutants also showed noticeable increase, from 8.8-fold to 13.2-fold, in the size of growth inhibition zone compared with that of the will type after treatment with cadmium. Therefore, the pigment production of A. agilis PY101 was found to decrease the toxic effects of cadmium to the bacterium.