Summary
The isolation and pure culture of the symbiotic nitrogen-fixing frankiae has always been difficult. In the past the isolation of these actinomycetes directly from soil samples has proven impossible and isolations from root nodules of many genera has been only poorly successful. We report here a modified sucrose fractionation procedure which increased the success of isolations from root nodules and which permitted the isolation ofFrankia directly from soil samples. Crushed nodule suspensions or soil suspensions were incubated briefly in 0.7% phenol (carbolic acid) just before application to a sucrose density gradient. This phenol incubation decreased the number of contaminating eubacteria and fungi but more importantly increased the number ofFrankia developing on the isolation plates. If the phenol incubation was used solely without sucrose fractionation noFrankia were isolated, suggesting the death of the organisms due to phenol toxicity. The use of selective nitrogen-deficient media proved important for the isolation of frankiae from soils.
Similar content being viewed by others
References
Baker D and Torrey J G 1979 The isolation and cultivation of actinomycetes root nodule endophytes.In Symbiotic Nitrogen Fixation in the Management of Temperate Forests. Eds. J C Gordon, C T Wheeler and D A Perry. Oregon State University Press, Corvallis, OR pp 38–56.
Baker D, Torrey J G and Kidd G H 1979 Isolation by sucrose-density fractionation and cultivationin vitro of actinomycetes from nitrogen-fixing root nodules. Nature London 281, 76–78.
Benson D 1982 Isolation ofFrankia strains from alder actinorhizal root nodules. Appl. Environ. Microbiol. 44, 461–465.
Berry A and Torrey J G 1979 Isolation and characterizationin vivo andin vitro of an actinomycetous endophyte fromAlnus rubra Bong,In Symbiotic Nitrogen Fixation in the Management of Temperate Forests. Eds. J C Gordon, C T Wheeler and D A Perry. Oregon State University Press, Corvallis, OR pp 69–83.
Diem H G, Gauthier D and Dommergues Y R 1982 Isolation ofFrankia from nodules ofCasuarina equisetifolia. Can. J. Microbiol. 28, 526–530.
Hoagland D R and Arnon D I 1950 The water culture method for growing plants without soil. Circ. Calif. Agric. Exp. Sta. 347 (revised edition).
Lalonde M 1979 A simple and rapid method for the isolation, cultivationin vitro and characterization ofFrankia strains fromAlnus root nodules.In Symbiotic Nitrogen Fixation in the Management of Temperate Forests. Eds. J C Gordon, C T Wheeler and D A Perry. Oregon State University Press, Corvallis, OR p 480.
Lalonde M, Calvert H E and Pine S 1981 Isolation and use ofFrankia strains in actinorhizae formation.In Current Perspectives in Nitrogen Fixation, Eds. A H Gibson and W E Newton, Aust. Acad. Sci., Canberra pp 296–299.
Lawrence C H 1956 A method of isolating actinomycetes from scabby potato tissue and soil with minimal contamination. Can. J. Bot. 34, 44–47.
Lechevalier M P, Baker D and Horriére F 1983 Physiology, chemistry, serology and infectivity of twoFrankia isolates fromAlnus incana. subsp.rugosa. Can. J. Microbiol. 61, 2826–2833.
Panthier J J, Diem H G and Dommergues Y 1979 Rapid method to enumerate and isolate soil actinomycetes antagonistic towards rhizobia. Soil Biol. Biochem. 11, 443–445.
Quispel A and Burggraaf A J P 1981Frankia, the diazotrophic endophyte from actinohizas.In Current Perspectives in Nitrogen Fixation. Eds. A H Gibson and W E Newton, Aust. Acad. Sci., Canberra pp 229–236.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Baker, D., O'Keefe, D. A modified sucrose fractionation procedure for the isolation of frankiae from actinorhizal root nodules and soil samples. Plant Soil 78, 23–28 (1984). https://doi.org/10.1007/BF02277837
Issue Date:
DOI: https://doi.org/10.1007/BF02277837