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Biology and Fertility of Soils

, Volume 46, Issue 4, pp 419–425 | Cite as

Competition for nodule occupancy between introduced and native strains of Rhizobium leguminosarum biovar trifolii

  • Andrea Rodríguez Blanco
  • Margarita Sicardi
  • Lillian Frioni
Short Communication

Abstract

The aim of this work was to evaluate the competitive ability between Rhizobium leguminosarum bv trifolii strain U204 used as commercial inoculants in Uruguay for Trifolium repens L. and Trifolium pratense L. and two native strains isolated from inoculated pastures of T. pratense. T126 is an efficient nitrogen fixer and a melanin producer strain; T70 is inefficient and a melanin non-producer strain; and U204 is very efficient in both hosts but is a melanin non-producer strain. Competitiveness between the strains was determined in experiments in pots and in growth pouches under controlled conditions. In the last experiment, we evaluated pH of plant nutrient solution and inoculum ratios. Plant dry weight was determined, and the identification of nodule bacteria was done using melanin production and DNA fingerprinting (GTG5-PCR). The U204 symbiotic efficiency was not affected by the co-inoculation with the others two native strains. The T70 strain was a poor competitor when was co-inoculated with one of the effective strains in both experiments. Our results confirmed a “selective nodulation” because an effective symbiosis occurred preferentially over an ineffective one in Trifolium species. The native effective strain competed with U204 for nodule formation in both clovers species, but the nodule occupancy depended on the inoculum ratio. The pH of nutritive solution did not affect competition ability of the studied strains. It may be possible to isolate efficient, competitive, and genetically different native rhizobial strains to be used as inoculant strains for clover pastures in Uruguay. Both (GTG)5-PCR and melanin production were useful methods to identify nodulating bacteria in competition studies.

Keywords

Competitiveness Melanin DNA fingerprinting Rhizobium leguminosarum bv trifolii Clover 

Notes

Acknowledgments

This work was partially financed by PEDECIBA (Programa de Desarrollo de Ciencias Básicas), Universidad de la República (UDELAR), Uruguay.

References

  1. Abdel-Fattah HI, Sonya HM, Sellim SM, Sharaf MS (2003) Use of random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) analysis to fingerprint some Rhizobium leguminosarum biovar viceae. Arab J Agric Sci 11:139–158Google Scholar
  2. Amarger N (1981) Selection of Rhizobium strains on their competitive ability for nodulation. Soil Biol Biochem 13:481–486CrossRefGoogle Scholar
  3. Beatiee GA, Clayton MK, Handelsman J (1989) Quantitative comparison of the laboratory and field competitiveness of Rhizobium leguminosarum bv phaseoli. Appl Environ Microbiol 55:2755–2761Google Scholar
  4. Beringer JE (1974) R factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198PubMedGoogle Scholar
  5. Brockwell J, Gault RR, Zorin M, Roberts MJ (1982) Effects of environmental variables on the competition between inoculum strains and naturalized populations of Rhizobium trifolium for nodulation of Trifolium subterraneum L and on rhizobia persistence in the soil. Austr J Agric Res 33:803–815CrossRefGoogle Scholar
  6. Brockwell J, Bottomley PJ, Thies JE (1995) Manipulation of rhizobia microflora for improving legume productivity and soil fertility: a critical assessment. Plant Soil 174:143–180CrossRefGoogle Scholar
  7. Castro S, Carrera I, Martínez-Drets G (2000) Methods to evaluate nodulation competitiveness between Sinorhizobium meliloti strains using melanin production as a marker. J Microbiol Meth 41:173–177CrossRefGoogle Scholar
  8. Chanway CP, Holl FB (1986) Suitable of intrinsic antibiotic resistance as a method of strain identification in Rhizobium trifolii. Plant Soil 93:287–291CrossRefGoogle Scholar
  9. Cubo TM, Buendía-Claveria AM, Beringer JE, Ruiz-Sainz JE (1988) Melanin production by Rhizboium strains. Appl Environ Microbiol 584:1812–1817Google Scholar
  10. Denton MD, Coventry DR, Howieson JG, Murphy PJ, Bellotti WD (2002) Competition between inoculated and native Rhizobium leguminosarum bv. trifolii for nodulation of annual clovers in alkaline soils. Austr J Agric Res 53:1019–1026CrossRefGoogle Scholar
  11. Diouf A, Spencer MM, Gueye M (2000) Use of the gus A gene marker in a competition study of the Rhizobium strains nodulating the common bean (Phaseolus vulgaris) in Senegal soils. World J Microbiol Biotechnol 16:337–340CrossRefGoogle Scholar
  12. Dowling DN, Broughton WJ (1986) Competition for nodulation of legumes. Ann Rev Microbiol 40:131–157CrossRefGoogle Scholar
  13. Dughri MH, Bottomley PJ (1983) Effect of acidity on the composition of an indigenous soil population of Rhizobium trifolii found in nodules of Trifolium subterraneum cultivar Mt. Barker. Appl Environ Microbiol 46:1207–1213PubMedGoogle Scholar
  14. Fabiano E, Arias A (1991) Competition between a native isolate of Rhizobium leguminosarum bv. trifolii and two commercial inoculant strains for nodulation of clover. Plant Soil 137:293–296CrossRefGoogle Scholar
  15. Gevers D, Huys G, Swings J (2001) Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205:31–36CrossRefPubMedGoogle Scholar
  16. Graham PH (1992) Stress tolerance in Rhizobium and Bradyrhizobium, and nodulation under adverse soil conditions. Can J Microbiol 38:475–484Google Scholar
  17. Hebb DM, Richardson AE, Reid R, Brockwell J (1998) PCR as an ecological tool to determine the establishment and persistence of Rhizobium strains introduced into field as seed inoculant. Austr J Agric Res 49:923–934CrossRefGoogle Scholar
  18. Howieson JG, Robson AD, Abbot LK (1992) Calcium modifies pH effects on the growth of acid-tolerant and acid-sensitive Rhizobium meliloti. Austr J Agric Res 43:765–772CrossRefGoogle Scholar
  19. Howieson JG, Ballard R (2004) Optimizing the legume symbiosis in stressful and competitive environments within southern Australia some contemporary thoughts. Soil Biol Biochem 36:1261–1273CrossRefGoogle Scholar
  20. Jhonston AWB, Hombrecher G, Brewin NJ, Cooper M (1982) Two transmissible plasmid in Rhizobium legunminosarum strains 300. J Gen Microbiol 128:85–93Google Scholar
  21. Jussila MM, Jurgens G, Lindstrom K, Suominen L (2006) Genetic diversity of culturable bacteria in oil-contaminated rhizosphere of Galega orientalis. Environ Pollution 139:244–257CrossRefGoogle Scholar
  22. Labandera C (1973) Competition between introduced and native Uruguayan strains of Rhizobium trifolii. Thesis M. Sci. School of Microbiology. University of New South Wales, p 66Google Scholar
  23. Labandera C, Vincent JM (1975) Competition between and introduced strain and native uruguayan strains of Rhizobium trifolii. Plant Soil 42:327–347CrossRefGoogle Scholar
  24. Marqués Pinto C, Yao PY, Vincent JM (1974) Nodulation competitiveness among strains of Rhizobium meliloti and R. trifolii. Austr J Agric Res 25:317–329CrossRefGoogle Scholar
  25. McDermott TR, Graham PH (1990) Competitive ability and efficiency in nodulte formation of strains of Bradyrhizobium japonicum. Appl Environ Microbiol 56:3035–3039Google Scholar
  26. Mrabet M, Mhamdi R, Tajini F, Tiwari R, Trabelsi M, Elarbi Aouani M (2005) Competitiveness and symbiotic effectiveness of a R. gallicum strain isolated from root nodules of Phaseolus vulgaris. Europ J Agron 22:209–216CrossRefGoogle Scholar
  27. Nick G, Jussila MM, Hoste B, Niemi RM, Kaijalainen S, de Lajudie P, Gillis M, de Bruijn FJ, Lindstrom K (1999) Rhizobia isolated from root nodules of tropical leguminous trees characterized using DNA-DNA dot-blot hybridisation and rep- PCR genomic fingerprinting. Syst Appl Microbiol 22:287–299Google Scholar
  28. Pryor HN, Lowther WL (2004) Caucasian clover rhizobia are not a threat to nitrogen fixation by white clover. Proc New Zeal Grass Assoc 66:285–289Google Scholar
  29. Ramirez ME, Israel DW, Wollum AG (1998) Using spontaneous antibiotic-resistant mutants to assess competitiveness of bradyrhizobial inoculants for nodulation of soybean. Can J Microbiol 44:753–758CrossRefGoogle Scholar
  30. Reuhs BL, Stephens SB, Geller DP, Kim JS, Glenn J, Przytycki J, Ojanew-Reuhs T (1999) Epitope identification for a panel of anti-Sinorhizobium meliloti monoclonal antibodies and application to the analysis of the K-antigen and lipopolysaccharides from bacteroids. Appl Environ Microbiol 65:5186–5191PubMedGoogle Scholar
  31. Rodríguez Blanco A (2005) Caracterización simbiótica, fisiológica y genética de rizobios aislados de praderas de Trifolium repens y T. pratense. Thesis M. Sci. Microbiología. PEDECIBA. Universidad de la República, Uruguay, p 105Google Scholar
  32. Rodríguez Blanco A, Csukasi F, Abreu C, Sicardi M (2008) Characterization of rhizobia from Sesbania species native to seasonally wetland areas in Uruguay. Biol Fertil Soils 44:925–932CrossRefGoogle Scholar
  33. Sargent L, Huang SZ, Rolfe BG, Djordjevic MA (1987) Split-root assays using Trifolium subterraneum show that Rhizobium infection induces a systemic response that can inhibit nodulation of another invasive Rhizobium strain. Appl Environ Microbiol 53:1611–1619PubMedGoogle Scholar
  34. SAS (1996) Statistical analysis system user’s guide. Statistics, 6th edn. SAS Institute Inc, CaryGoogle Scholar
  35. Schwieger F, Dammann-Kalinowski T, Dresing U, Selbitschka W, Munch JC, Puhler A, Keller M, Tebbe CC (2000) Field lysimeter investigation with luciferase-gene (luc)-tagged Sinorhizobium meliloti strains to evaluate the ecological significance of soil inoculation and a recA-mutation. Soil Biol Biochem 32:859–868CrossRefGoogle Scholar
  36. Selenska-Pobell (1994) How to monitor released rhizobia. Plant Soil 166:187–191CrossRefGoogle Scholar
  37. Sessitsch A, Jjemba PK, Hardarson G, Akkermans ADL, Wilson KJ (1997) Measurement of the competitiveness index of Rhizobium tropici strain CIAT899 derivatives marked with the gusA gene. Soil Biol Biochem 29:1099–1110CrossRefGoogle Scholar
  38. Stephens PM, Cooper JE (1988) Variation in speed of infection of no root hair zone of white clover and nodulating competitiveness among strains of Rhizobium trifolii. Soil Biol Biochem 20:465–476CrossRefGoogle Scholar
  39. Streit W, Botero L, Werner D, Beck D (1995) Competition for nodule occupancy on Phaseolus vulgaris by Rhizobium etli and Rhizobium tropici strains can be efficiently monitored in an ultisol during the early stages of growth using a constitutive GUS gene fusion. Soil Biol Biochem 27:1075–1081CrossRefGoogle Scholar
  40. Svenning MM, Gudmundsson J, Fagerli IL, Leinonen P (2001) Competition for nodule occupancy between introduced strains of Rhizobium leguminosarum biovar trifolii and its influence on plant production. Ann Bot 88:781–787CrossRefGoogle Scholar
  41. Thies JE, Bohlool BB, Singleton PW (1992) Environmental effects on nodulation for nodule occupancy between introduced and indigenous rhizobia and among introduced strains. Can J Microbiol 38:493–500CrossRefGoogle Scholar
  42. Triplett E, Sadowsky M (1992) Genetic of competition for nodulation of legumes. Ann Rev Microbiol 46:399–428CrossRefGoogle Scholar
  43. Versalovic J, Schneider M, de Bruijn FJ, Lupski J (1994) Genomic fingerprinting of bacteria using repetitive sequence based on PCR. Meth Mol Cells Biol 5:25–40Google Scholar
  44. Vincent JM (1970) A manual for the practical study of root-nodule bacteria. IBP Handbook N° 15. Blackwell, Oxford, p 164Google Scholar
  45. Wilson KJ (1995) Molecular techniques for the study of Rhizobial ecology in field. Soil Biol Biochem 27:501–514CrossRefGoogle Scholar
  46. Wilson KJ, Parra A, Botero L (1999) Application of the GUS marker gene technique to high-throughput screening of rhizobial competition. Can J Microbiol 45:678–685CrossRefGoogle Scholar
  47. Yates RJ, Howieson JG, Real D, Reeve WG, Vivas-Marfisi A, O’Hara GW (2005) Evidence of selection for effective nodulation in the Trifolium spp. symbiosis with Rhizobium leguminosarum biovar trifolii. Austr J Exp Agric 45:189–198CrossRefGoogle Scholar
  48. Yates RJ, Howieson JG, Reeve WG, Brau L, Speijers J, Nandasena K, Real D, Sezmis E, O’Hara GW (2008) Host-strain mediated selection for an effective nitrogen-fixing symbiosis between Trifolium spp. and Rhizobium leguminosarum biovar trifolii. Soil Biol Biochem 40:822–833CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Andrea Rodríguez Blanco
    • 1
  • Margarita Sicardi
    • 2
  • Lillian Frioni
    • 1
  1. 1.Microbiología, Facultad de AgronomíaUniversidad de la RepúblicaMontevideoUruguay
  2. 2.Microbiología de Suelos, Centro de Investigaciones Nucleares, Facultad de CienciasUniversidad de la RepúblicaMontevideoUruguay

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