Genotypic and symbiotic diversity studies of rhizobia nodulating Acacia saligna in Tunisia reveal two novel symbiovars within the Rhizobium leguminosarum complex and Bradyrhizobium |
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| Institution: | 1. Laboratory of Legumes and Sustainable Agroecosystems, Centre of Biotechnology of Borj-Cédria, BP 901 Hammam-lif 2050, Tunisia;2. Agriculture and Agri-Food Canada, 4902 Victoria Avenue North, Vineland Station, Ontario L0R 2E0, Canada;3. Faculty of Sciences, Centre de Biotechnologies Végétale et Microbienne, Biodiversité et Environnement, Mohammed V University, Rabat, Morocco;4. Agriculture and Agri-Food Canada, 960 Carling Avenue, Ottawa, Ontario K1A 0C6, Canada;5. Agroécologie, Institut Agro Dijon, CNRS, Univ. Bourgogne, INRAE, Univ. Bourgogne Franche-Comté, F-21000 Dijon, France;6. Laboratory of Rhizobia Biotechnology and Plant Breeding, University Oran1, Es Senia 31000, Algeria;1. Embrapa Soja, C.P. 231, 86001-970, Londrina, Paraná, Brazil;2. Department of Biotechnology, Universidade Estadual de Londrina, C.P. 10011, 86057-970, Londrina, Paraná, Brazil;3. Centre for Rhizobium Studies (CRS), Murdoch University, 90 South St. Murdoch, WA, Australia;1. Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidin, CSIC, Granada, Spain;2. Departamento de Microbiología y Genética, Universidad de Salamanca, Salamanca, Spain;3. Unidad Asociada Universidad de Salamanca-CSIC “Interacciones Planta-Microorganismo”, Spain;4. IRNASA-CSIC, Salamanca,Spain;1. Research Unit Biodiversity and Valorization of Arid Areas Bioresources (BVBAA) — Faculty of Sciences of Gabès, Erriadh, Zrig 6072, Tunisia;2. Centro de Biotecnología y Genómica de Plantas (UPM-INIA) and E.T.S.I. Agronómica, Alimentaria y de Biosistemas, Campus de Montegancedo, Universidad Politécnica de Madrid, 28223 Pozuelo de Alarcón, Madrid, Spain;3. Instituto de Ciencias Agrarias, CSIC, 28006, Madrid, Spain;1. Laboratory of Biodiversity and Valorization of Arid Areas Bioresources (BVBAA) – Faculty of Sciences of Gabes, Erriadh, Zrig 6072, Tunisia;2. Centro de Biotecnología y Genómica de Plantas (UPM-INIA), ETSI Agronómica, Alimentaria y de Biosistemas, Campus de Montegancedo, Universidad Politécnica de Madrid, Madrid, Spain;1. Centre de Biotechnologies Végétale et Microbienne, Biodiversité et Environnement, Faculté des Sciences, Université Mohammed V de Rabat, 4, Avenue Ibn Battouta, Rabat, Morocco;2. Centre de Recherche Forestière, Département des Eaux et Forêts, Avenue Omar Ibn El Khattab, BP 763, Rabat-Agdal 10050, Morocco;3. Faculté Polydiciplinaire, Université Sultan Moulay Slimane, Beni Mellal, Morocco;4. Departamento de Microbiología del Suelo y Sistemas Simbióticos, CSIC Estación Experimental del Zaidín, Apartado Postal 419, 18080 Granada, Spain;1. State Key Lab of Agrobiotechnology, Ministry of Agriculture Key Lab of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China;2. Escuela Nacional de Ciencias Biologicas, Instituto Politecnico Nacional, 11340 Mexico D. F., Mexico;3. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China |
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| Abstract: | Acacia saligna is an invasive alien species that has the ability to establish symbiotic relationships with rhizobia. In the present study, genotypic and symbiotic diversity of native rhizobia associated with A. saligna in Tunisia were studied. A total of 100 bacterial strains were selected and three different ribotypes were identified based on rrs PCR-RFLP analysis. Sequence analyses of rrs and four housekeeping genes (recA, atpD, gyrB and glnII) assigned 30 isolates to four putative new lineages and a single strain to Sinorhizobium meliloti. Thirteen slow-growing isolates representing the most dominant IGS (intergenic spacer) profile clustered distinctly from known rhizobia species within Bradyrhizobium with the closest related species being Bradyrhizobium shewense and Bradyrhizobium niftali, which had 95.17% and 95.1% sequence identity, respectively. Two slow-growing isolates, 1AS28L and 5AS6L, had B. frederekii as their closest species with a sequence identity of 95.2%, an indication that these strains could constitute a new lineage. Strains 1AS14I, 1AS12I and 6AS6 clustered distinctly from known rhizobia species but within the Rhizobium leguminosarum complex (Rlc) with the most closely related species being Rhizobium indicum with 96.3% sequence identity. Similarly, the remaining 11 strains showed 96.9 % and 97.2% similarity values with R. changzhiense and R. indicum, respectively. Based on nodC and nodA phylogenies and cross inoculation tests, these 14 strains of Rlc species clearly diverged from strains of Sinorhizobium and Rlc symbiovars, and formed a new symbiovar for which the name sv. “salignae” is proposed. Bacterial strains isolated in this study that were taxonomically assigned to Bradyrhizobium harbored different symbiotic genes and the data suggested a new symbiovar, for which sv. “cyanophyllae” is proposed. Isolates formed effective nodules on A. saligna. |
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| Keywords: | sv salignae sv cyanophyllae New symbiovars |
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