Multilocus sequence analysis of Brazilian Rhizobium microsymbionts of common bean (Phaseolus vulgaris L.) reveals unexpected taxonomic diversity |
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| Authors: | Renan Augusto Ribeiro Fernando Gomes Barcellos Fabiano L. Thompson Mariangela Hungria |
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| Affiliation: | 1. Embrapa Soja, Cx. Postal 231, 86001-970 Londrina, Paraná, Brazil;2. Universidade Estadual de Londrina, Department of Microbiology, Cx. Postal 60001, 86051-990 Londrina, Paraná, Brazil;3. UFRJ, Center of Health Sciences, Institute of Biology, Cx. Postal 68011, 21944-970 Rio de Janeiro, Rio de Janeiro, Brazil;1. Embrapa Soja, Cx. Postal 231, 86001-970 Londrina, Paraná, Brazil;2. Universidade Estadual de Londrina, Department of Microbiology, Cx. Postal 60001, 86051-990 Londrina, Paraná, Brazil;3. UFRJ, Center of Health Sciences, Institute of Biology, Cx. Postal 68011, 21944-970 Rio de Janeiro, Rio de Janeiro, Brazil;1. BNF and Stress Biology Laboratory, Department of Botany, Jai Narain Vyas University, Jodhpur 342001, Rajasthan, India;2. The James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK;3. Division of Plant Sciences, University of Dundee at the James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK;1. Departamento de Bioquímica, Microbiología, Biología Celular y Genética, Universidad de La Laguna, 38071 La Laguna, Tenerife, Spain;2. Departamento de Microbiología y Genética, Universidad de Salamanca, 37007 Salamanca, Spain;1. Julius Kühn-Institut, Federal Research Centre for Cultivated Plants (JKI), Institute for Epidemiology and Pathogen Diagnostics, Messeweg 11-12, 38104 Braunschweig, Germany;2. Research Institute of Horticulture, ul. Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland;3. Univ Lyon, UCBL, CNRS, INRA, VetAgo Sup, Ecologie Microbienne (LEM), 69622 Villeurbanne, France;1. Key Laboratory of Coastal Biology and Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, 264003 Yantai, China;2. Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, 11340 Mexico City, D.F., Mexico;3. Dongying Institute of Agriculture Sciences, 257000 Dongying, China;1. Laboratory of General and Agricultural Microbiology, Department of Crop Science, Agricultural University of Athens, Iera Odos 75, Votanikos, 11855 Athens, Greece;2. Laboratory of Vegetable Production, Department of Crop Science, Agricultural University of Athens, Iera Odos 75, Votanikos, 11855 Athens, Greece |
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| Abstract: | The diazotrophic bacteria collectively known as “rhizobia” are important for establishing symbiotic N2-fixing associations with many legumes. These microbes have been used for over a century as an environmentally beneficial and cost-effective means of ensuring acceptable yields of agricultural legumes. The most widely used phylogenetic marker for identification and classification of rhizobia has been the 16S rRNA gene; however, this marker fails to discriminate some closely related species. In this study, we established the first multilocus sequence analysis (MLSA) scheme for the identification and classification of rhizobial microsymbionts of common bean (Phaseolus vulgaris L.). We analyzed 12 Brazilian strains representative of a collection of over 850 isolates in addition to type and reference rhizobial strains, by sequencing recA, dnaK, gltA, glnII and rpoA genes. Gene sequence similarities among the five type/reference Rhizobium strains which are symbionts of common bean ranged from 95 to 100% for 16S rRNA, and from 83 to 99% for the other five genes. Rhizobial species described as symbionts of common bean also formed separate groups upon analysis of single and concatenated gene sequences, and clusters formed in each tree were in good mutual agreement. The five additional loci may thus be considered useful markers of the genus Rhizobium; in addition, MLSA also revealed broad genetic diversity among strains classified as Rhizobium tropici, providing evidence of new species. |
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