Bacterial flora of rhizosphere of the rice plant was examined to determine the relationship of nitrogen-fixing bacteria to rice plants. Of 7,344 bacterial strains from the rhizosphere, 1,005 strains (13.7%) reduced acetylene. Representative 74 strains with acetylene reduction activity were selected for identification. All of these strains were gram negative, aerobic or facultatively anaerobic, and most had ubiquinone (Q-10) and cellular fatty acid composition, mainly with straight-chain fatty acids. Twenty-two strains were assigned to the following known species: Azospirillum brasilense Tarrand, Krieg and Dobereiner (1 strain), Azospirillum lipoferum Tarrand, Krieg and Dobereiner (1 strain), Enterobacter cloacae (Jordon) Hormaeche and Edwards (4 strains), Erwinia herbicola (Lohnis) Dye (2 strains), Klebsiella oxytoca (Flugge) Lautrop (6 strains), and Xanthobacter autotrophicus (Baumgarten, Reh and Schlegel) Wiegel, Wilke, Baumgarten, Opitz and Schlegel (8 strains). Fifty-two strains could not be assigned to any known species.Since YOSHIDA and ANCAJAS (1) reported the association of free-living nitrogen-fixing bacteria with rice plants, several workers have isolated such bacteria and investigated the interrelation between bacteria and rice (2-17). Until recently, Azospirillum lipoferum (11-14), Azospirillium brasilense (12-14), Enterobacter cloacae (12,13,15), Klebsiella oxytoca (12, 13), Pseudomonas paucimobilis (12, 13), Pseudomonas sp. (16,17), and Clostridium sp. (5) have been isolated. SANO et al. (18) reported a seasonal variation of nitrogen-fixation in the rice plant and a possible symbiotic relation between bacteria and the plants. We have collaborated with them to investigate the relation between the nitrogen-fixing bacteria and the rice plant. This paper deals with the characterization and identification of the free-living nitrogen-fixing bacteria isolated from rice roots and paddy soil.