Inoculation density has a marked effect on nodulation and N fixation in soybean (Glycine max L.). Therefore, we conducted this study to determine the optimal inoculation density of Bradyhizobium japonicum SAY3-7 (SAY3-7) and Streptomyces griseoflavus P4 (P4) for plant growth, nodulation, and N fixation, and to investigate the effect of co-inoculation on selected soybean cultivars, using the optimal inoculation density. Nitrogen fixation, in terms of an acetylene reduction activity value, was measured using a flame ionization gas chromatograph. In this study, low-density single inoculation with P4 (10 5 or 10 6 cells mL -1 ) was associated with the highest plant biomass, compared with normal-and high-density single inoculation with P4 (10 7 or 10 8 cells mL -1 ). Moreover, low-density single or co-inoculations with SAY3-7 and/or P4 produced the highest nodule biomass and highest nitrogenase activity, compared with single or dual inoculation at other inoculation densities. Therefore, we evaluated low-density co-inoculation with P4 and SAY3-7, at the rate of 10 5 cells mL -1 , on selected soybean cultivars. Low-density co-inoculation increased the plant biomass, compared with un-inoculated plants. The effects of single and co-inoculation on nodulation did not differ significantly for any of the cultivars, except "Yezin-9" in the first experiment and "Shan Seine" in the second experiment. Low-density inoculation with both bacteria increased N fixation by 15% -75% for seven of the cultivars in the first experiment and by 15% -39% for three of the cultivars in the second experiment, compared with single inoculation with SAY3-7.
A. Z. Htwe, T. Yamakawa 1653Based on the overall results, we concluded that low-density co-inoculation with P4 and SAY3-7 gave improved plant growth and N fixation.