Abstract
The Azolla-Anabaena symbiosis has been used for centuries as a nitrogen biofertilizer in rice paddies. Genetic improvement of the symbiosis has been limited by the difficulty in identifying Azolla-Anabaena accessions and Anabaena azollae strains. The recently developed technique of DNA amplification fingerprinting (DAF) was applied to this problem. DAF uses single, short, oligonucleotide primers of arbitrary sequence to direct amplification of a characteristic set of DNA products by a thermostable DNA polymerase in a thermocycling reaction. The products are separated in polyacrylamide gels and detected by silver staining. DAF could easily distinguish and positively identify accessions of Azolla-Anabaena with DNA extracted from the intact symbioses. The contribution of prokaryotic Anabaena sequences to the fingerprint of the intact symbioses, however, ranged from 0 to 77%, depending on the primer sequence. Therefore, DNA extracted from the intact symbioses would not be suitable for Azolla taxonomy studies. The fingerprints of Anabaena strains isolated by sucrose gradient centrifugation from different species of Azolla could be easily distinguished, and DAF patterns were used to confirm the maternal pattern of transmission of Anabaena in a sexual hybrid. Template DNA extracted from roots was used to produce fingerprints for Azolla without interference from the microsymbiont. Comparison of the patterns from the parents and a hybrid gave strong evidence confirming sexual hybridization.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bassam BJ, Caetano-Anollés G, Gresshoff PM: Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal Biochem 196: 80–83 (1991).
Caetano-Anollés G, Bassam BJ, Gresshoff PM: DNA amplification fingerprinting using very short arbitrary oligonucleotide primers. Bio/technology 9: 553–557 (1991).
Caetano-Anollés G, Bassam BJ, Gresshoff PM: DNA amplification fingerprinting: a strategy for genome analysis. Plant Mol Biol Rep 9: 292–305 (1991).
Caetano-Anollés G, Bassam BJ, Gresshoff PM: Primer-template interactions during DNA amplification finger-printing with arbitrary oligonucleotides. Mol Gen Genet (in press).
Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19–21 (1983).
Dunham DG, Fowler K: Taxonomy and species recognition in Azolla Lam. In: Azolla Utilization, pp. 7–16, International Rice Research Institute, Los Baños, Philippines (1987).
Gebhardt JS, Nierzwicki-Bauer SA: Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria. Appl Environ Microbiol 57: 2141–2146 (1991).
Lin C, Liu Z, Zheng D, Tang L, Watanabe I: Re-establishment of symbiosis to Anabaena-free Azolla. Sci China (Ser B) 32: 551–559.
Liu CC, Chen Y, Tang L, Zheng Q, Song T, Chen M, Li Y, Lin T: Study on preparation and application of monoclonal antibodies to Anabaena azollae. Sci China (Ser B) 32: 562–569 (1989).
Peters GA, Mayne BC: The Azolla, Anabaena azollae relationship. I. Initial characterization of the association. Plant Physiol 53: 813–819 (1974).
Peters GA, Meeks JC: The Azolla-Anabaena symbiosis: basic biology. Annu Rev Plant Physiol Plant Mol Biol 40: 193–210 (1989).
Plazinski J: The Azolla-Anabaena symbiosis. In: Gresshoff PM (ed) The Molecular Biology of Symbiotic Nitrogen Fixation, pp. 51–75. CRC Press, Boca Raton, FL (1990).
Plazinski J, Croft L, Taylor R, Zheng Q, Rolfe BG, Gunning BES: Indigenous plasmids in Anabaena azollae: their taxonomic distribution and existence of regions of homology with symbiotic genes of Rhizobium. Can J Microbiol 37: 171–181 (1991).
Plazinski J, Franche C, Liu CC, Lin T, Shaw W, Gunning BES, Rolfe BG: Taxonomic status of Anabaena azollae: An overview. Plant Soil 108: 185–190 (1988).
Plazinski J, Zheng Q, Taylor R, Rolfe BG, Gunning BES: Use of DNA/DNA hybridization techniques to authenticate the production of new Azolla-Anabaena symbiotic associations. FEMS Microbiol Lett 65: 199–204 (1989).
Plazinski J, Zheng Q, Taylor R, Croft L, Rolfe BG, Gunning BES: DNA probes show genetic variation in cyanobacteria symbionts of the Azolla fern and a closer relationship to free-living Nostoc strains than to free-living Anabaena strains. Appl Environ Microbiol 56: 1263–1270 (1990).
Tan BC, Payawal P, Watanabe I, Lacdan N, Ramirez C: Modern taxonomy of Azolla: a review. Phil Agric 69: 4491–516 (1986).
Tuan DT, Thuyet TQ: Use of Azolla in rice production in Vietnam. In: Nitrogen and Rice, pp. 395–405. International Rice Research Institute, Los Baños, Philippines (1979).
VanCat D, Watanabe I, Zimmerman WJ, Lumpkin TA, deWaha Baillonville T: Sexual hybridization among Azolla species. Can J Bot 67: 3482–3485 (1989).
Watanabe I, Espinas CR, Berja NS, Alimagno BV: Utilization of the Azolla-Anabaena complex as a nitrogen fertilizer for rice. IRRI Research Paper Series. International Rice Research Institute 11: 1–6 (1977).
Watanabe I, Lin C, Ramirez C, Lapis MT, Santiago-Ventura T, Liu CC: Physiology and agronomy of Azolla-Anabaena symbiosis. In: Skinner FA et al., (eds) Nitrogen Fixation with Non-legumes, pp. 57–62. Kluwer Academic Publishers, Dordrecht (1989).
Watanabe I, Lin C, Santiago-Ventura T: Response to high temperature of the Azolla-Anabaena association, determined in both the fern and in the cyanobacterium. New Phytol 111: 625–630 (1989).
Wei WX, Jin GY, Zhang N: Preliminary report of Azolla hybridization studies (in Chinese). Bull Fujian Acad Agric Sci 1: 73–79 (1986).
Welsh J, McClelland M: Fingerprinting genomes using PCR with arbitrary primers. Nucl Acids Res 18: 7213–7218 (1990).
Williams JGK, Hanafey MK, Rafalski JA, Tingey SV: Genetic analysis using RAPD markers. In: Recombinant DNA Volume. Meth in Enzymol, in press (1992).
Williams JGK, Kubilek AR, Livak KJ, Rafalski JA, Tingey SV: DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18: 6531–6535 (1990).
Zimmerman WJ, Lumpkin TA, Watanabe I: Isozyme differentiation of Azolla Lam. Euphytica 42: 163–170 (1989).
Zimmerman WJ, Lumpkin TA, Watanabe I: Classification of Azolla spp., Section Azolla. Euphytica 43: 223–232 (1989).
Zimmerman WJ, Watanabe I, Lumpkin TA: The Anabaena-Azolla symbiosis: Diversity and relatedness of neotropical host taxa. Plant Soil 137: 167–170. (1991).
Zimmerman WJ, Watanabe I, Ventura T, Payawal P, Lumpkin TA: Aspects of the genetic and botanical status of neotropical Azolla species. New Phytol 119: 561–566. (1991).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Eskew, D.L., Caetano-Anollés, G., Bassam, B.J. et al. DNA amplification fingerprinting of the Azolla-Anabaena symbiosis. Plant Mol Biol 21, 363–373 (1993). https://doi.org/10.1007/BF00019951
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00019951