Free-living and symbiotic characteristics of chlorate resistant mutants of Rhizobium japonicum

Abstract

This work investigated the usefulness of chlorate resistance as a method for the selection of nitrate reductase negative (NR−) strains from Rhizobium japonicum (61A76) and evaluated the symbiotic characteristics of these strains. Chlorate resistant strains were selected from populations seeded on CS 7 agar containing 10 or 20 mM KClO3 and incubated in 2% air – 98% N2–CO2 (95:5).Over 200 resistant strains were isolated, 58% of which lacked the dissimilatory nitrate reductase. In 12 selected isolates, some strains had also lost the assimilatory nitrate reductase, but all retained hydrogenase activity.Chlorate resistant strains inoculated to soybean seedlings were equal to or better than the parent strain in terms of nodule mass and acetylene reduction. Those strains lacking both assimilatory and dissimilatory nitrate reductase showed the best symbiotic characteristics, suggesting that chlorate resistance in R. japonicum could be a useful method for the selection of strains with superior nitrogen-fixing characteristics.