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Applied and Environmental Microbiology, May 2001, p. 2116-2122, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2116-2122.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Diversity and Specificity of Frankia Strains in Nodules of Sympatric Myrica gale, Alnus incana, and Shepherdia canadensis Determined by rrs Gene Polymorphism

Valerie Huguet,1 Janet Mccray Batzli,2 Jeff F. Zimpfer,2 Philippe Normand,1 Jeffrey O. Dawson,2 and Maria P. Fernandez1,*

Laboratoire d'Ecologie Microbienne, UMR CNRS 5557, Université Claude Bernard Lyon 1, 69622 Villeurbanne Cedex, France,1 and Department of Natural Resources and Environmental Sciences, University of Illinois, Urbana, Illinois 618012

Received 25 October 2000/Accepted 28 February 2001

The identity of Frankia strains from nodules of Myrica gale, Alnus incana subsp. rugosa, and Shepherdia canadensis was determined for a natural stand on a lake shore sand dune in Wisconsin, where the three actinorhizal plant species were growing in close proximity, and from two additional stands with M. gale as the sole actinorhizal component. Unisolated strains were compared by their 16S ribosomal DNA (rDNA) restriction patterns using a direct PCR amplification protocol on nodules. Phylogenetic relationships among nodular Frankia strains were analyzed by comparing complete 16S rDNA sequences of study and reference strains. Where the three actinorhizal species occurred together, each host species was nodulated by a different phylogenetic group of Frankia strains. M. gale strains from all three sites belonged to an Alnus-Casuarina group, closely related to Frankia alni representative strains, and were low in diversity for a host genus considered promiscuous with respect to Frankia microsymbiont genotype. Frankia strains from A. incana nodules were also within the Alnus-Casuarina cluster, distinct from Frankia strains of M. gale nodules at the mixed actinorhizal site but not from Frankia strains from two M. gale nodules at a second site in Wisconsin. Frankia strains from nodules of S. canadensis belonged to a divergent subset of a cluster of Elaeagnaceae-infective strains and exhibited a high degree of diversity. The three closely related local Frankia populations in Myrica nodules could be distinguished from one another using our approach. In addition to geographic separation and host selectivity for Frankia microsymbionts, edaphic factors such as soil moisture and organic matter content, which varied among locales, may account for differences in Frankia populations found in Myrica nodules.

* Corresponding author. Mailing address: Laboratoire d'Ecologie Microbienne, UMR CNRS 5557, Université Claude Bernard Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne Cedex, France, Phone: 33 4-72-43-29-37. Fax: 33 4-72-43-12-23. E-mail: fernandz{at}biomserv.univ-lyonl.fr.

Applied and Environmental Microbiology, May 2001, p. 2116-2122, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2116-2122.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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