Azotobacter vinelandii strains UW (Nif+) or UW1 (Nif-) were induced to form competent cells by growing cultures in Burk medium without added iron. Competent cells were generated in either liquid or solid medium. The competent culture was highly colored by the characteristic fluorescent green pigment of A. vinelandii but all pigmented cultures were not competent. In liquid culture, competent cells required a fixed nitrogen source (ammonia, nitrate, or urea) that was also a nitrogenase repressor. Glucose, sucrose, glycerol, and mannitol (1% w/v) were excellent C-sources for competence induction while acetate (1% w/v) and beta-hydroxybutyrate (0.2% w/v) were ineffective. Cells induced in liquid culture remained competent for a 12-h period before the end of the exponential growth phase, then competence decreased rapidly upon the initiation of the stationary phase of growth.