Differential effects of coinoculations with Pseudomonas jessenii PS06 (a phosphate-solubilizing bacterium) and Mesorhizobium ciceri C-2/2 strains on the growth and seed yield of chickpea under greenhouse and field conditions

  • Angel Valverde
  • Araceli Burgos
  • Tiziana Fiscella
  • Raúl Rivas
  • Encarna Velázquez
  • Claudino Rodríguez-Barrueco
  • Emilio Cervantes
  • Manuel Chamber
  • José-Mariano Igual
Conference paper
Part of the Developments in Plant and Soil Sciences book series (DPSS, volume 102)


In the course of a project carried out in two regions of Spain, Castilla y León and Andalucía, aiming to find useful biofertilizers for staple grain-legumes, an efficient rhizobia nodulating chickpea (termed as C-2/2) and a powerful in vitro phosphate-solubilizing bacterial strain (termed as PS06) were isolated. Analyses of their 16S rDNA sequence indicated that they belong to the bacterial species Mesorhizobium ciceri and Pseudomonas jessenii, respectively. Greenhouse and field experiments were carried out in order to test the effect of single and dual inoculations on chickpea (ecotype ILC-482) growth. Under greenhouse conditions, plants inoculated with Mesorhizobium ciceri C-2/2 alone had the highest shoot dry weight. The inoculation treatment with P. jessenii PS06 yielded a shoot dry weight 14% greater than the uninoculated control treatment, but it was not correlated with shoot P contents. However, the co-inoculation of C-2/2 with PS06 resulted in a decrease in shoot dry weight with respect to the inoculation with C-2/2 alone. Under field conditions, plants inoculated with M. ciceri C-2/2, in single or dual inoculation, produced higher nodule fresh weight, nodule number and shoot N content than the other treatments. Inoculation with P. jessenii PS06 had no significant effect on plant growth. However, the co-inoculation treatment ranked the highest in seed yield (52% greater than the uninoculated control treatment) and nodule fresh weight. These data suggest that P. jessenii PS06 can act synergistically with M. ciceri C-2/2 in promoting chickpea growth. The contrasting results obtained between greenhouse and field experiments are discussed.

Key words

chickpea Mesorhizobium PGPR phosphate-solubilizing bacteria plant yield Pseudomonas 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alagawadi A R and Gaur A C 1992 Inoculation of Azospirillum brasilense and phosphate-solubilizing bacteria on yield of sorghum [Sorghum bicolor (L.) Moench] in dry land. Trop. Agric. 69, 347–350.Google Scholar
  2. Antoun H, Beauchamp C J, Goussard N, Chabot R and Lalande R 1998 Potential of Rhizobium and Bradyrhizobium species as growth promoting bacteria on non-legumes: effect on radishes (Raphanus sativus L.). Plant Soil 204, 57–67.CrossRefGoogle Scholar
  3. Belimov A A, Kojemiakov A P and Chuvarliyeva C V 1995 Interaction between barley and mixed cultures of nitrogen fixing and phosphate-solubilizing bacteria. Plant Soil 173, 29–37.CrossRefGoogle Scholar
  4. Bergersen F J 1961 The growth of Rhizobium in synthetic media. Aust. J. Biol. Sci. 14, 349–360.Google Scholar
  5. Bolton H, Elliot L F, Turco R F and Kennedy A C 1990 Rhizoplane colonization of pea seedlings by Rhizobium leguminosarum and a deleterious root colonizing Pseudomonas sp. and effects on plant growth. Plant Soil 123, 121–124.Google Scholar
  6. Chabot R, Antoun H and Cescas M P 1993 Growth stimulation of corn and romiane lettuce by microorganisms solubilizing inorganic phosphorous. Can. J. Microbiol. 39, 941–947.CrossRefGoogle Scholar
  7. Chabot R, Antoun H and Cescas M P 1996 Growth promotion of maize and lettuce by phosphate-solubilizing Rhizobium leguminosarum biovar phaseoli. Plant Soil 184, 311–321.CrossRefGoogle Scholar
  8. Chabot R, Beauchamp C J, Kloepper J W and Antoun H 1998 Effect of phosphorous on root colonization and growth promotion of maize by bioluminescent mutants of phosphate-solubilizing Rhizobium leguminosarum biovar. phaseoli. Soil Biol. Biochem. 30, 1615–1618.CrossRefGoogle Scholar
  9. Dashti N, Zhang F, Hynes R and Smith D L 1998 Plant growth-promoting rhizobacteria accelerate nodulation and increase nitrogen fixation activity by field grown soybean [Glycine max (L.) Merr.] under short season conditions. Plant Soil 200, 205–213.CrossRefGoogle Scholar
  10. Davison J 1988 Plant beneficial bacteria. Biotechnology 6, 282–286.CrossRefGoogle Scholar
  11. de Freitas J R, Banerjee M R and Germida J J 1997 Phosphate-solubilizing rhizobacteria enhance the growth and yield but not phosphorous uptake in canola (Brassica napus L.). Biol. Fertil. Soils 24, 358–364.CrossRefGoogle Scholar
  12. Glick B R 1995 The enhancement of plant growth by free-living bacteria. Can. J. Microbiol. 41, 109–117.CrossRefGoogle Scholar
  13. Gupta R, Singal R, Sankar A, Chander R M and Kumar R S 1994 A modified plate assay for screening phosphate solubilizing microorganisms. J. Gen. Appl. Microbiol. 40, 255–260.Google Scholar
  14. Halder A K, Mishra A K, Bhattacharyya P and Chakrabartty P K 1990 Solubilization of rock phosphate by Rhizhobium and Bradyrhizobium. J. Gen. Appl. Microbiol. 36, 81–92.Google Scholar
  15. Hirsch A M, Fang Y, Asad S and Kapulnik Y 1997 The role of phytohormones in plant-microbe symbioses. Plant Soil 194, 171–184.CrossRefGoogle Scholar
  16. Höfte M, Boelens J and Verstraete W 1991 Seed protection and promotion of seedling emergence by the plant growth beneficial Pseudomonas strains 7NSK2 and ANP15. Soil Biol. Biochem. 23, 407–410.CrossRefGoogle Scholar
  17. Igual J M, Valverde A, Cervantes E and Velázquez E 2001 Phosphate-solubilizing bacteria as inoculants for agriculture: use of updated molecular techniques in their study. Agronomie 21, 561–568.CrossRefGoogle Scholar
  18. Illmer P and Schinner F 1992 Solubilization of inorganic phosphates by microorganisms isolated from forest soil. Soil Biol. Biochem. 24, 389–395.CrossRefGoogle Scholar
  19. Kim K Y, Jordan D and McDonald G A 1998 Effect of phosphate-solubilizing bacteria and vesicular-arbuscular mycorrhizae on tomato growth and soil microbial activity. Biol. Fertil. Soils 26, 79–87.CrossRefGoogle Scholar
  20. Kloepper J W and Schroth M N 1978 Plant growth-promoting rhizobacteria on radishes. In Proceedings of the IV International Conference on Plant Pathogenic Bacteria, vol. 2. Eds. Gibert-Clarey and Tours. pp. 879–882. Station de Phatologie Végétale et Phytobactériologie, INRA, Angers, France.Google Scholar
  21. Kundu B S and Gaur A C 1984 Rice response to inoculation with N2-fixing and P-solubilizing microorganisms. Plant Soil 79, 227–234.CrossRefGoogle Scholar
  22. Liljeroth E, Bååth E, Mathiasson I and Lundborg T 1990a Root exudation and rhizoplane bacterial abundance of barley (Hordeum vulgare L) in relation to nitrogen fertilization and root growth. Plant Soil 127, 81–89.CrossRefGoogle Scholar
  23. Liljeroth E, Van Veen J A and Miller H J 1990b Assimilate translocation to the rhizosphere of two wheat lines and subsequent utilization by rhizosphere microorganisms at two nitrogen concentrations. Soil Biol. Biochem. 22, 1015–1021.CrossRefGoogle Scholar
  24. Marschner P, Gerendás J and Sattelmacher B 1999 Effect of N concentration and N source on root colonization by Pseudomonas fluorescens 2-79RLI. Plant Soil 215, 135–141.CrossRefGoogle Scholar
  25. McLaughlin M J, Alston A M and Martin J K 1988 Phosphorus cycling in wheat-pasture rotations II The role of the microbial biomass in phosphorus cycling. Aust. J. Soil Res. 26, 333–342.CrossRefGoogle Scholar
  26. Oberson A, Friesen D K, Rao I M, Bühler S and Frossard E 2001 Phosphorus transformations in an Oxisol under contrasting land-use systems: The role of the soil microbial biomass. Plant Soil 237, 197–210.CrossRefGoogle Scholar
  27. Oehl F, Oberson A, Probst M, Fliessbach A, Roth H R and Frossard E 2001 Kinetics of microbial phosphorus uptake in cultivated soils. Biol. Fertil. Soils 34, 31–41.CrossRefGoogle Scholar
  28. O’Sullivan D J and O’Gara F 1992 Traits of fluorescent Pseudomonas spp. involved in suppression of plant root pathogens. Microbiol. Rev. 56, 662–676.PubMedGoogle Scholar
  29. Pal S S 1998 Interaction of an acid tolerant strain of phosphate solubilizing bacteria with a few acid tolerant crops. Plant Soil 198, 169–177.CrossRefGoogle Scholar
  30. Pearson W R and Lipman D J 1988 Search for DNA homologies was performed with the FASTA program. Proc. Natl. Acad. Sci. USA 85, 2444–2448.PubMedCrossRefGoogle Scholar
  31. Peix A, Rivas-Boyero A A, Mateos P F, Rodríguez-Barrueco C, Martínez-Molina E and Velázquez E 2001a Growth promotion of chickpea and barley by a phosphate solubilizing strain of Mesorhizobium mediterraneum under growth chamber conditions. Soil Biol. Biochem. 33, 103–110.CrossRefGoogle Scholar
  32. Peix A, Mateos P F, Rodríguez-Barrueco C, Martínez-Molina E and Velázquez E 2001b Growth promotion of common bean (Phaseolus vulgaris L.) by a strain of Burkholderia cepacia under growth chamber conditions. Soil Biol. Biochem. 33, 1927–1935.CrossRefGoogle Scholar
  33. Persello-Cartieaux F, Nussaume L and Robaglia C 2003 Tales from the underground: molecular plant-rhizobacteria interactions. Plant Cell Environ. 26, 189–199.CrossRefGoogle Scholar
  34. Piccini D and Azcón R 1987 Effect of phosphate-solubilizing bacteria and vesicular arbuscular mycorrhizal (VAM) on the utilization of bayoran rock phosphate by alfalfa plants using a sand-vermiculite medium. Plant Soil 101, 45–50.CrossRefGoogle Scholar
  35. Ray J, Bagyaraj D J and Manjunath A 1981 Influence of soil inoculation with versicular arbuscular mycorrhizal (VAM) and a phosphate dissolving bacteria on plant growth and 32P uptake. Soil Biol. Biochem. 13, 105–108.CrossRefGoogle Scholar
  36. Rigaud J and Puppo A 1975 Indole-3-acetic catabolism by soybean bacteroids. J. Gen. Microbiol. 88, 223–228.Google Scholar
  37. Rivas R, Velázquez E, Valverde A, Mateos P F and Martínez-Molina E 2001 A two primers random amplified polymorphic DNA procedure to obtain polymerase chain reaction fingerprints of bacterial species. Electrophoresis 22, 1086–1089.PubMedCrossRefGoogle Scholar
  38. Rodríguez H and Fraga R 1999 Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnol. Adv. 17, 319–339.PubMedCrossRefGoogle Scholar
  39. Sarawgi S K, Tiwari P K and Tripathi R S 1999 Uptake and balance sheet of nitrogen and phosphorus in gram (Cicer arietinum) as influenced by phosphorus, biofertilizers and micronutrients under rainfed condition. Indian J. Agron. 44, 768–772.Google Scholar
  40. Schmelz E A, Engelberth J, Alborn H T, O’Donnell P, Sammons M, Toshima H and Tumlinson J H III 2003 Simultaneous analysis of phytohormones, phytotoxins, and volatile organic compounds in plants. Proc. Natl. Acad. Sci. USA 100, 10552–10557.PubMedCrossRefGoogle Scholar
  41. Seong K Y, Hofte M, Boelens J and Verstraete W 1991 Growth, survival and root colonization of plant growth beneficial Pseudomonas fluorescens ANP15 and Pseudomonas aeruginosa 7NSK2 at different temperatures. Soil Biol. Biochem. 23, 423–428.CrossRefGoogle Scholar
  42. Sindhu S S, Gupta S K and Dadarwal K R 1999 Antagonistic effect of Pseudomonas spp. on pathogenic fungi and enhancement of growth of green gram (Vigna radiata). Biol. Fertil. Soils 29, 62–68.CrossRefGoogle Scholar
  43. Sindhu S S, Suneja S, Goel A K, Parma N and Dadarwal K R 2002 Plant growth promotion effects of Pseudomonas sp. on coinoculation with Mesorhizobium sp. Cicer strain under sterile and “wilt sick”’ soil conditions. Appl. Soil Ecol. 19, 57–64CrossRefGoogle Scholar
  44. Snedecor G W and Cochran W G 1989 Statistical Methods. Iowa State University Press, Ames, Iowa 503 pp.Google Scholar
  45. Subba Rao N S 1993 Biofertilizers in Agriculture and Forestry. Oxford and IBH Publishing Co. Pvt. Ltd, New Delhi 242 pp.Google Scholar
  46. Tan K H 1996 Soil Sampling, Preparation, and Analysis. Marcel Dekker, Inc, New York 408 pp.Google Scholar
  47. Thompson J D, Gibson T J, Plewniak F, Jeanmougin F and Higgins D G 1997 The clustalX windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acid Res. 24, 4876–4882.CrossRefGoogle Scholar
  48. Tomar R K S, Namdeo K N and Ranghu J S 1996 Efficacy of phosphate solubilizing bacteria biofertilizers with phosphorus on growth and yield of gram (Cicer arietinum). Indian J. Agron. 41, 412–415.Google Scholar
  49. Toro N, Azcón R and Barea J M 1997 Improvement of arbuscular mycorrhiza development by inoculation of soil with phosphate-solubilizing rhizobacteria to improve rock phosphate bioavailability (32P) and nutrient cycling. Appl. Environ. Microbiol. 63, 4408–4412.PubMedGoogle Scholar
  50. Toro N, Azcón R and Barea JM 1998 The use of isotopic dilution techniques to evaluate the interactive effects of Rhizobium genotype, mycorrhizal fungi, phosphate-solubilizing rhizobacteria and rock phosphate on nitrogen and phosphorus acquisition by Medicago sativa. New Phytol. 138, 265–273.CrossRefGoogle Scholar
  51. Umrit G and Friesen D K 1994 The effect of C:P ratio of plant residues added to soils of contrasting phosphate sorption capacities on P uptake by Panicum maximum (Jacq.). Plant Soil 158, 275–285.CrossRefGoogle Scholar
  52. Vincent J M 1970 The cultivation, isolation and maintenance of rhizobia. In A Manual for the Practical Study of Root-Nodule. Ed. J M Vincent. pp. 1–13. Blackwell Scientific Publications, Oxford.Google Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  • Angel Valverde
    • 1
  • Araceli Burgos
    • 2
  • Tiziana Fiscella
    • 1
    • 4
  • Raúl Rivas
    • 3
  • Encarna Velázquez
    • 3
  • Claudino Rodríguez-Barrueco
    • 1
  • Emilio Cervantes
    • 1
  • Manuel Chamber
    • 2
  • José-Mariano Igual
    • 1
  1. 1.Instituto de Recursos Naturales y Agrobiología-CSICSalamancaSpain
  2. 2.CIFA Las Torres-TomejilAlcalá del Rio, SevillaSpain
  3. 3.Departamento de Microbiología y GenéticaUniversidad de SalamancaSalamancaSpain
  4. 4.Agriculture FacultyUniversity of CataniaCataniaItaly

Personalised recommendations