Cultivable heterotrophic N₂-fixing bacterial diversity in rice fields in the Yangtze River Plain

Heterotrophic N₂-fixing bacteria are a potentially important source of N₂ fixation in rice fields due to the moist soil conditions. This study was conducted at eight sites along a geographic gradient of the Yangtze River Plain in central China. A nitrogen-free solid malate-sucrose medium was used to isolate heterotrophic N₂-fixing bacteria. Numbers of the culturable N₂-fixing bacteria expressed as CFU (colony forming units) ranged between 1.41±0.42×106 and 1.24±0.23×10⁸ in the sampled paddy field sites along the plain. Thirty strains with high ARA (acetylene reduction activity) were isolated and purified; ARA of the strains varied from 0.9 to 537.8 nmol C₂H₄ culture^-1 h^-1, and amounts of ¹⁵N fixed ranged between 0.008 and 0.4866 mg·culture^-1·day^-1. According to morphological and biochemical characteristics, 14 strains were identified as the genus Bacillus, 2 as Burkholderia, 1 as Agrobacterium, 4 as Pseudomonas, 2 as Derxia, 1 as Alcaligenes, 1 as Aeromonas, 2 as Citrobacter, and 3 strains belonged to the corynebacterform group.