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Rhizobium cellulosilyticum as a co-inoculant enhances Phaseolus vulgaris grain yield under greenhouse conditions

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Abstract

The Rhizobium-legume symbiosis is a complex partnership with many factors, with initial bacterial colonization of the plant root surface and primary infection as key early stages. Two molecules are strongly involved in these processes: the structural carbohydrate cellulose and the enzyme cellulase, which breaks down the former and allows rhizobia to infect the roots. Here, we report the effect on common bean (Phaseolus vulgaris L.) after co-inoculation of the non-nodulating, cellulase-overproducing strain Rhizobium cellulosilyticum ALA10B2T and the P. vulgaris-nodulating R. leguminosarum strain TPV08. In order to elucidate the effect of combined inoculation with both strains, we designed greenhouse assays, including single inoculation with strain TPV08, co-inoculation with both strains and an uninoculated treatment in non-sterile peat. Chemical fertilizers were not added. Chlorophyll content in the leaves was measured after the flowering stage by spectrophotometry and was considered to be indicative of the nutrient status of the plants. Nodule formation was observed on roots of the inoculated plants, while no nodulation was observed on roots of the uninoculated plants. The results indicate a synergistic effect between the two Rhizobium strains. Co-inoculated plants exhibited significant increases in seed yield and nitrogen content in comparison with the uninoculated control plants and with plants inoculated with a single strain. It is suggested that co-inoculation with strain ALA10B2T greatly increased the efficiency of N fixation by strain TPV08.

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Acknowledgments

This work was supported by the Spanish Government project AGL2011-29227. Alexandra Díez-Méndez was supported by a PhD fellowship from Junta de Castilla y León (Regional Government).

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Authors and Affiliations

  1. Department of Microbiology and Genetics. Edificio Departamental de Biología, Lab 213. Universidad de Salamanca (USAL), Plz. Dres. de la Reina s/n, 37007, Salamanca, Spain

    Alexandra Diez-Mendez, Esther Menéndez, Lorena Celador-Lera, Raúl Rivas & Pedro F. Mateos

  2. Instituto Hispano Luso de Investigaciones Agrarias (CIALE), Salamanca, Spain

    Alexandra Diez-Mendez, Esther Menéndez, Lorena Celador-Lera & Pedro F. Mateos

  3. Institute of Microbiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20, Praha 4, Prague, Czech Republic

    Paula García-Fraile

  4. Associated Unit USAL-CSIC (IRNASA), Salamanca, Spain

    Raúl Rivas & Pedro F. Mateos

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  1. Alexandra Diez-Mendez
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  2. Esther Menéndez
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  3. Paula García-Fraile
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  4. Lorena Celador-Lera
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  5. Raúl Rivas
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  6. Pedro F. Mateos
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Correspondence to Pedro F. Mateos.

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ESM 1

Qualitative assays of cellulose (A) and cellulase (B) production. Cellulose production was detected using Congo Red staining, and CMCase activity by double-layer plate assays after inoculation with 10 μL of each strain. 1. R. rhizogenes ATCC11325T, 2. R. cellulosilyticum ALA10B2T, 3. R. leguminosarum bv phaseoli TPV08, 4. R. pisi DSM30132T, 5. R. jaguaris CCGE2052T, 6. R. radiobacter ATCC19358T, 7. R. multihospitium LMG3946T, 8. R. vignae LMG25447T, 9. R. endophyticum CCGE525T, 10. R. selenitireducens LMG24075T, 11. R. leguminosarum ATCC10004T, 12. R. indigoferae CCBAU710492T. (GIF 51 kb)

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Diez-Mendez, A., Menéndez, E., García-Fraile, P. et al. Rhizobium cellulosilyticum as a co-inoculant enhances Phaseolus vulgaris grain yield under greenhouse conditions. Symbiosis 67, 135–141 (2015). https://doi.org/10.1007/s13199-015-0372-9

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