Rhizobium strain identification in Arachis hypogaea nodules by enzyme-linked immunosorbent assay (ELISA)

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Abstract
The technique of ELISA was used for serological identification of peanut Rhizobium strains both in cell suspension of pure culture and in single root nodules of groundnut (A. hypogaea) plants. Antisera of 3 peanut Rhizobium strains were tested against 8 different Rhizobium isolates. Three serogroups identified by agglutination and immunodiffusion tests were confirmed by ELISA. In this experiment ELISA was more sensitive by 4-6 orders of magnitude than the agglutination and immunodiffusion tests and enabled the detection of Rhizobium antigens in cell suspensions of 104-105 cells/ml. The reactions of culture and nodule antigens were identical for all strains investigated. ELISA enabled the precise typing of rhizobial isolates in single small root nodules. The minimum fresh weight of nodule tissue necessary to perform the ELISA test was 0.4 mg crushed in 1 ml of phosphate-buffered saline (PBS). ELISA was also successfully used for strain identification in mixed inoculated plants. One of the strains in each pair formed most of the nodules examined.