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Characterization of Rhizobium Japonicum Strains Isolated from the Nodules of Soybean

F. Begum
BCSRP, Bangladesh Agricultural Research Council, Dhaka.

S. A. Ahmed
Microbiology Section, Food Science and Technology Division, BCSIR Laboratories, Dhaka.

G. K. Joarder
Microbiology Section, Food Science and Technology Division, BCSIR Laboratories, Dhaka.

Abstract/ Executive Summary:

A considerable number of strains have been isolated from the nodules of soybean plants grown in this country. For their identification morphological. biochemical & physiological studies were performed. Strain varieties were identified. It was found that the isolates differ much in their biochemical reaction confirming the diversity of the strain characters among the isolates. The effectiveness of a selected number of strains were also been tested by nodule formation using a simplified method. They were found also to differ in infective properties to seme extent although the overall results of formation of nodules were encouraging.

Keywords: Nodules of Soybean, Rhizobium Japonicum, Characterization

Location: BCSRP, Bangladesh Agricultural Research Council, Dhaka.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Soybean

Objectives:

To the carried out on the isolation and characterization of the strains of Rhizobium japonicum from the nodules of soybean cultivated here and assesment of their nodulation efficiency are reported.

Methodology:

Isolation and selection of cultures: The method described by Vincent (1970) was generally followed in the isolation of the strains of Rhizobium species from the nodules of soybean using yeastmannitol agar with congo red. The surface sterelized nodules were creuashed and streaked on agar plates aseptically. Colonies which did not take congo red and grown rather late (about 4-5 days) having typical colonial properties of Rhizobium species were isolated purified and tested for other characteristics of Rhizobium species.

Culture maintenance: Rhizobiul cultures are maintained on yeast mannitol agar slopes at referigerator temperature (10^oc). All organisms were checked for purity at least everyone month and before usinq in any experiments.

Tests were carried out on cultures grown for 3-7 days on yeast mannitol agar slants. Gram staining was done by usual method. Sudanophlic granules were tested with sudan Black-B and counter staining withsafranin. Colonical features were determined from the cultures grown on the 7 days yeast mannitol agar medium. Inoculated plates of yeast mannitol, agar were incubated at 4⁰C and 39⁰C for 7 days and the growth was noted. The pH of the tubes of liquid yeast mannitol agar slant were adjusted as require and after sterilization by the addition of dil. HCI and NaOH as required. Inoculated tubes were incubated for 7 days and the growth was observed visually and microscopically.

The biochemical properties such as (i) Catalase activity (ii) Reduction of nitrate (iii) Reduction of methylene blue and (iv) Tolerance of sodium chloride were done according to the methods followed by Graham, P. H. and Parker (1964) (V) Growth in litmus milk: Growth in litmus milk was carried out according to Soc. Amer. Bacteriologist (1957) (vi) Oxidase activity: Oxidase activity was tested according to Kovaks (1956). (vii) H₂S production: H₂S production was determined according to Hunter and Crecelius. H. G. (1938). (viii) Voges Proskauer test was done according to Soc. of Amer Bact (1957). (ix) Urease production was done according to the method described. (x) The strains were inoculated in Norris medium and observed for growth in nitrogen free medium.

A simplified process was developed to test the nodulation by the strains in general way. Garden soil and sand in the proportion 3: 1 were steriIized in oven then put into the pots (approximate diameter about 25cm). Sterile tap water was used to soak the soil. The soybeans were surface sterilized as mention above then smeared with the strains. The seeds were sowed 3-4 cm below the surface of the soil in the pots. The pots were kept in darkness for 3-5 days for proper germination then put into sunshine. The formation of nodules were observed after three weeks. Blanks with uninoculated seeds were also run.

Source of Information: Bangladesh Journal of Agricultural Vol. 5 No.3

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

It was found that the isolates differ much in their biochemical reaction confirming the diversity of the strain characters among the isolates. The effectiveness of a selected number of strains were also been tested by nodule formation using a simplified method. They were found also to differ in infective properties to seme extent although the overall results of formation of nodules were encouraging.

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