Abstract
To appraise the usefulness of the enzyme-linked immunosorbent assay (ELISA) technique for examining the serological diversity of slow-growing rhizobia, twelve diverse strains from three countries were examined with four antisera. Eleven of the strains were from the “cowpea” miscellany, and the twelfth was a Rhizobium japonicum strain. Some “cowpea” strains showed no antigenic relatedness with each other while others were closely related, and some showed a greater affinity with the R. japonicum strain than with other “cowpea” strains. All of the strains showed antigenic homology to an isolate from a wild Arachis sp., while two strains isolated from adjacent plants of the same cultivar had little homology. These patterns ofrelatedness and diversity clearly demonstrated the utility of the ELISA method, and so it was used to examine 53 strains isolated from cowpeas grown at three West African locations, Maradi (Niger), Ibadan and Onne (Nigeria). Broad ranges of serological diversity were found in the rhizobia at each location, moreover each population had its own general characteristics. Maradi strains were highly reactive with the five antisera used, Onne strains less so, and Ibadan strains even less so. ELISA reactivity correlated with colony morphology but not with nodulation potential.
Similar content being viewed by others
References
Ahmad MH, Eaglesham ARJ, Hassouna S, Seaman B, Ayanaba A, Mulongoy K, Pulver EL (1981) Examining the potential for inoculant use with cowpeas in West African soils. Trop Agric (Trin) 58:325–335
Berger JA, May SN, Berger LR, Bohlool BB (1979) Colorimetric enzymelinked immunosorbent assay for the identification of strains of Rhizobium in culture and in the nodules of lentils. Appl Environ Microbiol 37:642–646
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem 72:248–254
Bushnell OA, Sarles WB (1939) Investigations upon the antigenic relationships among the root-nodule bacteria of the soybean, cowpea, and lupine cross-inoculation groups. J Bacteriol 38:401–410
Damirgi SM, Frederick LR, Anderson IC (1967) Serogroups of Rhizobium japonicum in soybean nodules as affected by soil types. Agron J 59:10–12
Dudman WF (1964) Immune diffusion analysis of the extracellular soluble antigens of two strains of Rhizobium meliloti. J Bacteriol 88:783–794
Engvall E, Perlmann P (1972) Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol 109:129–135
Garvey JS, Cremer NE, Sussdorf DH (1977) Methods in immunology. 3rd ed. W. A. Benjamin Inc., Mass., USA
Graham PH (1963) Antigenic affinities of the root nodule bacteria of legumes. Antonie van Leeuwenhoek 29:281–291
Ikram A, Broughton WJ (1980) Rhizobia in tropical legumes. VIII. Serological characteristics of Posphocarpus tetragonolobus (L.) DC. isolates. Soil Biol Biochem 12:83–87
Jones DG, Russell PE (1972) The application of immunofluorescence techniques to host plant nodule bacteria selectivity experiments using Trifolium repens. Soil Biol Biochem 4:277–282
Kishinevsky B, Bar-Joseph M (1979) Rhizobium strain identification in Arachis hypogaea nodules by enzyme-linked immunosorbent assay (ELISA). Can J Microbiol 24:1537–1543
Means UM, Johnson HW, Date RA (1964) Quick serological method of classifying strains of Rhizobium japonicum in nodules. J Bacteriol 87:457–553
Schmidt EL (1973) Fluorescent-antibody techniques for the study of microbial ecology. Bull Ecol Res Comm (Stockholm) 17:67–76
Sinha RC, Peterson EA (1980) Homologous serological analysis of Rhizobium meliloti strains by immunodiffusion. Can J Microbiol 26:1157–1161
Trinick MJ (1969) Identification of legume nodule bacteroids by the fluorescent antibody reaction. J Appl Bacteriol 32:181–186
Vincent JM (1970) A manual for the practical study of root nodule bacteria. IBP Handbook No 15. Blackwell Scientific Publications, Oxford
Vincent JM, Humphrey B (1970) Taxonomically significant group antigens in Rhizobium. J Gen Microbiol 63:379–382
Vincent JM, Humphrey B, Skrdleta V (1973) Group antigens in slowgrowing rhizobia. Arch Mikrobiol 89:79–82
Voller A, Bidwell D, Bartlett A (1973) Microplate enzyme immunoassays for the immunodiagnosis of virus infections. In: Rose NR, Friedman H (eds) Manual of clinical immunology. Amer Soc Microbiol, Washington, DC, pp 506–512
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ahmad, M.H., Eaglesham, A.R.J. & Hassouna, S. Examining serological diversity of “cowpea” rhizobia by the ELISA technique. Arch. Microbiol. 130, 281–287 (1981). https://doi.org/10.1007/BF00425941
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00425941