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Applied and Environmental Microbiology, September 2000, p. 3987-3997, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genotypic Characterization of Bradyrhizobium Strains Nodulating Small Senegalese Legumes by 16S-23S rRNA Intergenic Gene Spacers and Amplified Fragment Length Polymorphism Fingerprint Analyses

Florence Doignon-Bourcier,1 Anne Willems,2 Renata Coopman,2 Gisele Laguerre,3 Monique Gillis,2 and Philippe de Lajudie1,*

Laboratoire des Symbioses Tropicales et Méditerranéennes, I. R. D., Campus de Baillarguet, 34398 Montpellier Cedex 5,1 and Laboratoire de Microbiologie des Sols, CMSE, INRA, B.V. 1540, 21034 Dijon Cedex,3 France, and Laboratorium voor Microbiologie, Universiteit Ghent, K.-L. Ledeganckstraat, 35, B-9000, Ghent, Belgium2

Received 27 March 2000/Accepted 11 July 2000

We examined the genotypic diversity of 64 Bradyrhizobium strains isolated from nodules from 27 native leguminous plant species in Senegal (West Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia, which play an ecological role and have agronomic potential in arid regions. The strains were characterized by intergenic spacer (between 16S and 23S rRNA genes) PCR and restriction fragment length polymorphism (IGS PCR-RFLP) and amplified fragment length polymorphism (AFLP) fingerprinting analyses. Fifty-three reference strains of the different Bradyrhizobium species and described groups were included for comparison. The strains were diverse and formed 27 groups by AFLP and 16 groups by IGS PCR-RFLP. The sizes of the IGS PCR products from the Bradyrhizobium strains that were studied varied from 780 to 1,038 bp and were correlated with the IGS PCR-RFLP results. The grouping of strains was consistent by the three methods AFLP, IGS PCR-RFLP, and previously reported 16S amplified ribosomal DNA restriction analysis. For investigating the whole genome, AFLP was the most discriminative technique, thus being of particular interest for future taxonomic studies in Bradyrhizobium, for which DNA is difficult to obtain in quantity and quality to perform extensive DNA:DNA hybridizations.

* Corresponding author. Mailing address: Laboratoire des Symbioses Tropicales et Méditerranéennes, I. R. D., Campus de Baillarguet, 34398 Montpellier Cedex 5, France. Phone: (33) 04 67 61 58 00, ext. 4218. Fax: (33) 04 67 59 38 02. E-mail: P-De.Lajudie{at}mpl.ird.fr.

Applied and Environmental Microbiology, September 2000, p. 3987-3997, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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