Estimation of Root Nodule Development by Rhizobium sp Using Hydroponic Culture

A nitrogen-fixing bacterium isolated from the root nodules of a cultivated leguminous plant, soybean (Glycine max L.), was cultivable and was identified as Rhizobium sp. Bacterial species isolated from root nodules of wild leguminous plants including -bush clover, white dutch clover, wisteria, and false acacia were identified as Burkholderia cepacia, Pseudomonas migulae, Pseudomonas putida, and Flavobacterium sp, respectively, all of which are heterotrophic bacteria that grow in the rhizosphere. Temperature gradient gel electrophoresis (TGGE) 16S-rDNA bands extracted directly from the bacterial population within the root nodules of the wild leguminous plants were identified as Rhizobium sp, Mesorhizobium sp, and Bradyrhizobium sp. none were cultivable. Rhizobium sp. isolated from soybean root nodule generated approximately 48 and 19 mg/L of ammonium in glucose- and starch-defined medium, respectively, during 8 days of growth. The growth rate of Rhizobium sp. was increased by the addition of yeast extract but not by the addition of ammonium. Km and V,m for starch saccharification measured with the extracellular crude enzyme of Rhizobium sp. were 0.7556 mg/L and 0.1785 mg/L/min, respectively. The inoculation of Rhizobium sp. culture into a hydroponic soybean plant culture activated root nodule development and soybean plant growth. The inoculated Rhizobium sp. survived for at least 4 weeks, based on the TGGE pattern of 16S-rDNA. The 16S-rDNA of Rhizobium sp. isolated from newly developed root nodules was homologous with the inoculated species.