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Large-Scale Transposon Mutagenesis of Photosynthetic Bradyrhizobium Sp. Strain ORS278 Reveals New Genetic Loci Putatively Important for Nod-Independent Symbiosis with Aeschynomene indica

June 2010 , Volume 23 , Number  6
Pages  760 - 770

Katia Bonaldi,1,2 Benjamin Gourion,1 Joel Fardoux,1 Laure Hannibal,1 Fabienne Cartieaux,1 Marc Boursot,1 David Vallenet,3 Clémence Chaintreuil,1 Yves Prin,4 Nico Nouwen,1 and Eric Giraud1

1Laboratoire des Symbioses Tropicales et Méditerranéennes, IRD, UMR-IRD/SupAgro/INRA/UM2/CIRAD, F-34398 Montpellier, France; 2UM2, F-34398 Montpellier, France; 3Genoscope, CNRS-UMR 8030, Atelier de Génomique Comparative, F-91057 Evry, France; 4Laboratoire des Symbioses Tropicales et Méditerranéennes, CIRAD, UMR-IRD/SupAgro/INRA/UM2/CIRAD, F-34398 Montpellier, France

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Accepted 23 February 2010.

Photosynthetic Bradyrhizobium strains possess the unusual ability to form nitrogen-fixing nodules on a specific group of legumes in the absence of Nod factors. To obtain insight into the bacterial genes involved in this Nod-independent symbiosis, we screened 15,648 Tn5 mutants of Bradyrhizobium sp. strain ORS278 for clones affected in root symbiosis with Aeschynomene indica. From the 268 isolated mutants, 120 mutants were altered in nodule development (Ndv--) and 148 mutants were found to be deficient in nitrogen fixation (Fix--). More than 50% of the Ndv-- mutants were found to be altered in purine biosynthesis, strengthening the previous hypothesis of a symbiotic role of a bacterial purine derivative during the Nod-independent symbiosis. The other Ndv-- mutants were auxotrophic for pyrimidines and amino acids (leucine, glutamate, and lysine) or impaired in genes encoding proteins of unknown function. The Fix-- mutants were found to be affected in a wide variety of cellular processes, including both novel (n = 56) and previously identified (n = 31) genes important in symbiosis. Among the novel genes identified, several were involved in the Calvin cycle, suggesting that CO2 fixation could play an important role during this symbiosis.

© 2010 The American Phytopathological Society