Comparative studies of nitrogen-fixing bacteria associated with grasses in Israel with Azospirillum brasilense

Free-living, dinitrogen-fixing bacteria associated with roots of grasses were isolated from several locations in Israel. Bacteria with characteristics similar to those of Azospirillum were isolated from Cynodon dactylon roots and were compared with Azospirillum brasilense from Brasil (Sp-7) and from California (Cd). Colonies of the Israeli isolates were yellow and consisted of curved rods, 0.5-0.6 micron in diameter with polar flagella, whereas colonies of A. brasilense were pink (Sp-7) and red (Cd) and the cells were 1.0-1.1 micron in diameter with polar flagella. Ultraviolet absorption spectra of soluble c and membrane-bound c and b cytochromes were similar in all isolates. When grown in semisolid agar medium with or without ammonium chloride all isolates formed a growth zone below the surface. However, they grew best under aerobic conditions in liquid medium containing NH4Cl. All isolates could use salts of malate and lactate, arabinose, and galactose, but not mannitol, as sole carbon sources; they did not need biotin to shorten their lag phase. One Israeli isolate was capable of growing and fixing nitrogen with glucose as a sole carbon source. The Israeli isolates formed aggregates above pH 7.6 in liquid or semisolid medium and were capable of reducing nitrate to nitrogen gas under anaerobic conditions.