DIVERSITY AND HOST SPECIFICITY OF AZOLLA CYANOBIONTS

[en] A unique, hereditary symbiosis exists between the water fern Azolla and cyanobacteria that reside within a cavity in the dorsal leaf-lobe of the plant. This association has been studied extensively, and questions have frequently been raised regarding the number and diversity of cyanobionts (cyanobacterial symbionts) among the different Azolla strains and species. In this work, denaturating gradient gel electrophoresis (DGGE) and a clone library based on the 16S rRNA gene were used to study the genetic diversity and host specificity of the cyanobionts in 35 Azolla strains covering a wide taxonomic and geographic range. DNA was extracted directly from the cyanobacterial packets, isolated after enzymatic digestion of the Azolla leaves. Our results indicated the existence of different cyanobiont strains among Azolla species, and diversity within a single Azolla species, independent of the geographic origin of the host. Furthermore, the cyanobiont exhibited host-species specificity and showed most divergence between the two sections of genus Azolla, Azolla and Rhizosperma. These findings are in agreement with the recent redefinition of the taxon Azolla cristata within the section Azolla. With regard to the taxonomic status of the cyanobiont, the genus Anabaena of the Nostocaceae family was identified as the closest relative by this work.

Disciplines :

Phytobiology (plant sciences, forestry, mycology...)
Microbiology

Author, co-author :

Papaefthimiou, Dimitra

Van Hove, Charles

Lejeune, André

Rasmussen, Ulla

Wilmotte, Annick ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie

Language :

English

Title :

DIVERSITY AND HOST SPECIFICITY OF AZOLLA CYANOBIONTS

Publication date :

2008

Journal title :

Journal of Phycology

ISSN :

0022-3646

eISSN :

1529-8817

Publisher :

Blackwell Publishing, Oxford, United Kingdom

Volume :

Pages :

60-70

Peer reviewed :

Peer Reviewed verified by ORBi

Name of the research project :

ESF Programme CYANOFIX

Funders :

ESF - European Science Foundation [FR]

Available on ORBi :

since 23 July 2009

Statistics

Number of views

83 (2 by ULiège)

Number of downloads

7 (3 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. Lipman, D. J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25 : 3389 402.
Baker, J. A., Entsch, B. McKay, D. B. 2003. The cyanobiont in an Azolla fern is neither Anabaena nor Nostoc. FEMS Microbiol. Lett. 229 : 43 7.
Bonfield, J. K., Beal, K. F., Betts, M. J. Staden, R. 2002. Trev: a DNA trace editor and viewer. Bioinformatics (Oxf.) 18 : 194 5.
Boutte, C., Grubisic, S., Balthasart, P. Wilmotte, A. 2006. Testing of primers for the study of cyanobacterial molecular diversity by DGGE. J. Microbiol. Methods 65 : 542 50.
Braun-Howland, E. B. Nierzwicki-Bauer, S. A. 1990. Azolla-Anabaena symbiosis: biochemistry, physiology, ultra-structure and molecular biology. In Rai, A. N. Ed Handbook of Symbiotic Cyanobacteria. CRC Press Inc., Boca Raton, Florida, pp. 65 117.
Caudales, R., Wells, J. M., Antoine, A. D. Butterfield, J. E. 1995. Fatty acid composition of symbiotic cyanobacteria from different host plant (Azolla) species: evidence for co-evolution of host and symbiont. Int. J. Syst. Bacteriol. 45 : 364 70.
Caudales, R., Wells, J. M., Butterfield, J. E. Antoine, A. D. 1993. Host related variability in fatty acid composition of symbiotic cyanobacteria isolated from the aquatic fern Azolla. In Sato, S., Ishida, M. Ishikawa, H. Eds Endocytobiology V - Proceedings of the 5th International Colloquium on Endocytobiology and Symbiosis. Tubingen University Press, Tübigen, Germany, pp. 261 8.
Cole, J. R., Chai, B., Marsh, T. L., Farris, R. J., Wang, Q., Kulam, S. A., Chandra, S. et al., 2003. The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy. Nucleic Acids Res. 31 : 442 3.
Evrard, C. Van Hove, C. 2004. Taxonomy of the American Azolla species (Azollaceae): a critical review. Syst. Geogr. Plants 74 : 301 18.
Felsenstein, J. 1993. PHYLIP (Phylogeny Inference Package) version 3.5c. Distributed by the author. Department of Genetics, University of Washington, Seattle.
Gebhardt, J. S. Nierzwicki-Bauer, S. A. 1991. 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 6.
Gugger, M., Lyra, C., Henriksen, P., Coute, A., Humbert, J. F. Sivonen, K. 2002. Phylogenetic comparison of the cyanobacterial genera Anabaena and Aphanizomenon. Int. J. Syst. Evol. Microbiol. 52 : 1867 80.
Hall, T. A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 41 : 95 8.
Hoagland, D. R. Arnon, D. I. 1950. The Water-Culture Method for Growing Plants without Soil. Calif. AES Circular 347. University of California, Berkeley, California.
Huelsenbeck, J. P. Ronquist, F. 2001. MRBAYES: Bayesian inference of phylogeny. Bioinformatics (Oxf.) 17 : 754 5.
Jackson, C. R., Roden, E. E. Churchill, P. F. 2000. Denaturing gradient gel electrophoresis can fail to separate 16S rDNA fragments with multiple base differences. Mol. Biol. Today 1 : 49 51.
Kim, J. J. H., Krawczyk, K., Lorentz, W. P. Zimmerman, W. J. 1997. Fingerprinting cyanobionts and hosts of the Azolla symbiosis by DNA amplification. World J. Microbiol. Biotechnol. 13 : 97 101.
Komárek, J. Anagnostidis, K. 1989. Modern approach to the classification system of cyanophytes. 4-Nostocales. Arch. Hydrobiol. Suppl. 82 : 247 345.
Ladha, J. K. Watanabe, I. 1982. Antigenic similarity among Anabaena azollae separated from different species of Azolla. Biochem. Biophys. Res. Commun. 109 : 675 82.
Lechno-Yossef, S. Nierzwicki-Bauer, S. A. 2002. Azolla-Anabaena symbiosis. In Rai, A. N., Bergman, B. Rasmussen, U. [Eds.] Cyanobacteria in Symbiosis. Kluwer Academic Publishers, Dordrecht, the Netherlands, pp. 153 78.
Liu, Z. Z., Cheng, Y. Q., Tang, L. F., Zheng, Q., Song, T. Y., Chen, M. A., Li, Y. Y. Lin, T. L. 1986. Studies on monoclonal antibodies against Anabaena azollae. J. Fujian Acad. Agric. Sci. 1 : 1 6.
Liu, Z. Z., Cheng, Y. Q., Tang, L. F., Zheng, Q., Song, T. Y., Chen, M. A., Li, Y. Y. Lin, T. L. 1989. Study on preparation and application of monoclonal antibodies to Anabaena azollae. Sci. China Ser. B 32 : 560 9.
Lyra, C., Suomalainen, S., Gugger, M., Vezie, C., Sundman, P., Paulin, L. Sivonen, K. 2001. Molecular characterization of planktic cyanobacteria of Anabaena, Aphanizomenon, Microcystis and Planktothrix genera. Int. J. Syst. Evol. Microbiol. 51 : 513 26.
McCowen, S. M., Mac Arthur, L. Gates, J. E. 1987. Azolla fern lectins that specifically recognize endosymbiotic cyanobacteria. Curr. Microbiol. 14 : 329 33.
Meeks, J. C., Joseph, C. M. Haselkorn, R. 1988. Organization of the nif genes in cyanobacteria in symbiotic association with Azolla and Anthoceros. Arch. Microbiol. 150 : 61 71.
Moore, L. R., Rocap, G. Chisholm, S. W. 1998. Physiology and molecular phylogeny of co-existing Prochlorococcus ecotypes. Nature (Lond.) 393 : 464 7.
Neefs, J. M., Van de Peer, Y., Hendriks, L. De Wachter, R. 1990. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. 18 : 2237 317.
Nierzwicki-Bauer, S. A. Haselkorn, R. 1986. Differences in mRNA levels in Anabaena living freely or in symbiotic association with Azolla. EMBO J. 5 : 29 35.
Nübel, U., Garcia-Pichel, F. Muyzer, G. 1997. PCR primers to amplify 16S rRNA genes from cyanobacteria. Appl. Environ. Microbiol. 63 : 3327 32.
Pabby, A., Prasanna, R., Nayak, S. Singh, P. K. 2003. Physiological characterization of the cultured and freshly isolated endosymbionts from different species of Azolla. Plant Physiol. Biochem. (Paris) 41 : 73 9.
Page, R. D. M. 1996. TREEVIEW: an application to display phylogenetic trees on personal computers. Comput. Appl. Biosci. 12 : 357 8.
Peters G. A., Jr., Toia, R. E., Raveed, D. Levine, N. J. 1978. The Azolla-Anabaena azollae relationship. VI. Morphological aspects of the association. New Phytol. 80 : 583 93.
Plazinski, J., Zheng, Q., Taylor, R., Croft, L., Rolfe, B. G. Gunning, B. E. S. 1990. DNA probes show genetic variation in cyanobacterial symbionts of the Azolla fern and a closer relationship to the free living Nostoc strains than to free living Anabaena strains. Appl. Environ. Microbiol. 56 : 1263 70.
Posada, D. Crandall, K. A. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics (Oxf.) 14 : 817 8.
Rajaniemi, P., Hrouzek, P., Kastovska, K., Willame, R., Rantala, A., Hoffmann, L., Komárek, J. Sivonen, K. 2005. Phylogenetic and morphological evaluation of the genera Anabaena, Aphanizomenon, Trichormus and Nostoc (Nostocales, Cyanobacteria). Int. J. Syst. Evol. Microbiol. 55 : 11 26.
Reid, J. D., Plunkett, G. M. Peters, G. A. 2006. Phylogenetic relationships in the heterosporous fern genus Azolla (Azollaceae) based on DNA sequence data from three noncoding regions. Int. J. Plant Sci. 167 : 529 38.
Rocap, G., Distel, D. L., Waterbury, J. B. Chisholm, S. W. 2002. Resolution of Prochlorococcus and Synechococcus ecotypes by using 16S-23S ribosomal DNA internal transcribed spacer sequences. Appl. Environ. Microbiol. 68 : 1180 91.
Saunders, R. M. K. Fowler, K. 1992. A morphological taxonomic revision of Azolla Lam. section Rhizosperma (Mey.) Mett. (Azollaceae). Bot. J. Linn. Soc. 109 : 329 57.
Saunders, R. M. K. Fowler, K. 1993. The supraspecific taxonomy and evolution of the fern genus Azolla (Azollaceae). Plant Syst. Evol. 184 : 175 93.
Schloss, P. D. Handelsman, J. 2005. Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl. Environ. Microbiol. 71 : 1501 6.
Stackebrandt, E. Göbel, B. M. 1994. Taxonomic note: a place for DNA-DNA re-association and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol. 44 : 846 9.
Strasburger, E. 1884. Die Controversen der indirecten Kemtheilung. Arch. Mikr. Anat. 23 : 301.
Svenning, M. M., Eriksson, T. Rasmussen, U. 2005. Phylogeny of symbiotic cyanobacteria within the genus Nostoc based on 16S rDNA sequence analyses. Arch. Microbiol. 183 : 19 26.
Swofford, D. L. 2003. PAUP*: Phylogenetic Analysis Using Parsimony (*and Other Methods), Version 4. Sinauer Associates, Sunderland, Massachusetts.
Taton, A., Grubisic, S., Brambilla, E., De Wit, R. Wilmotte, A. 2003. Cyanobacterial diversity in natural and artificial microbial mats of Lake Fryxell (McMurdo Dry Valleys, Antarctica): a morphological and molecular approach. Appl. Environ. Microbiol. 69 : 5157 69.
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. Higgins, D. G. 1997. The CLUSTALX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25 : 4876 82.
Tomaselli, L., Margheri, M. C., Giovannetti, L., Sili, C. Carlozzi, P. 1988. The taxonomy of Azolla spp. cyanobionts. Ann. Microbiol. 38 : 157 61.
Tomitani, A., Knoll, A. H., Cavanaugh, C. M. Terufumi, O. 2006. The evolutionary diversification of cyanobacteria: molecular-phylogenetic and paleontological perspectives. Proc. Natl. Acad. Sci. U. S. A. 103 : 5442 7.
Van Coppenolle, B., McCough, S. R., Watanabe, I., Huang, N. Van Hove, C. 1995. Genetic diversity and phylogeny analysis of Anabaena azollae based on RFLRs detected in Azolla-Anabaena azollae DNA complexes using nif gene probes. Theor. Appl. Genet. 91 : 589 97.
Watanabe, I., Roger, P. A., Ladha, J. K. Van Hove, C. 1992. Biofertilizer Germplasm Collections at IRRI. The International Rice Research Institute, Los Banos, Philippines, 66 pp.
van Wintzingerode, F., Gölbel, U. B. Stackebrandt, E. 1997. Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis. FEMS Microbiol. Rev. 21 : 213 29.
Zheng, W. W., Nilsson, M., Bergman, B. Rasmussen, U. 1999. Genetic diversity and classification of cyanobacteria in different Azolla species by the use of PCR fingerprinting. Theor. Appl. Genet. 99 : 1187 93.