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Most Acid-Tolerant Chickpea Mesorhizobia Show Induction of Major Chaperone Genes upon Acid Shock

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Abstract

Our goals were to evaluate the tolerance of mesorhizobia to acid and alkaline conditions as well as to investigate whether acid tolerance is related to the species or the origin site of the isolates. In addition, to investigate the molecular basis of acid tolerance, the expression of chaperone genes groEL and dnaKJ was analyzed using acid-tolerant and sensitive mesorhizobia. Tolerance to pH 5 and 9 was evaluated in liquid medium for 98 Portuguese chickpea mesorhizobia belonging to four species clusters. All isolates showed high sensitivity to pH 9. In contrast, mesorhizobia revealed high diversity in terms of tolerance to acid stress: 35 % of the isolates were acid sensitive and 45 % were highly tolerant to pH 5 or moderately acidophilic. An association between mesorhizobia tolerance to acid conditions and the origin soil pH was found. Furthermore, significant differences between species clusters regarding tolerance to acidity were obtained. Ten isolates were used to investigate the expression levels of the chaperone genes by northern hybridization. Interestingly, most acid-tolerant isolates displayed induction of the dnaK and groESL genes upon acid shock while the sensitive ones showed repression. This study suggests that acid tolerance in mesorhizobia is related to the pH of the origin soil and to the species cluster of the isolates. Additionally, the transcriptional analysis suggests a relationship between induction of major chaperone genes and higher tolerance to acid pH in mesorhizobia. This is the first report on transcriptional analysis of the major chaperones genes in mesorhizobia under acidity, contributing to a better understanding of the molecular mechanisms of rhizobia acidity tolerance.

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References

  1. Alexandre A, Brígido C, Laranjo M, Rodrigues S, Oliveira S (2009) A survey of chickpea rhizobia diversity in Portugal reveals the predominance of species distinct from Mesorhizobium ciceri and Mesorhizobium mediterraneum. Microb Ecol 58:930–941

    Article  PubMed  Google Scholar 

  2. Alexandre A, Laranjo M, Oliveira S (2006) Natural populations of chickpea rhizobia evaluated by antibiotic resistance profiles and molecular methods. Microb Ecol 51:128–136

    Article  PubMed  CAS  Google Scholar 

  3. Alexandre A, Oliveira S (2011) Most heat-tolerant rhizobia show high induction of major chaperone genes upon stress. FEMS Microbiol Ecol 75:28–36

    Article  PubMed  CAS  Google Scholar 

  4. Amarger N, Macheret V, Laguerre G (1997) Rhizobium gallicum sp. nov. and Rhizobium giardinii sp. nov., from Phaseolus vulgaris nodules. Int J Syst Bacteriol 47:996–1006

    Article  PubMed  CAS  Google Scholar 

  5. Angelini J, Taurian T, Morgante C, Ibanez F, Castro S, Fabra A (2005) Peanut nodulation kinetics in response to low pH. Plant Physiol Biochem 43:754–759

    Article  PubMed  CAS  Google Scholar 

  6. Ausubel FM, Brent R, Kingston RE, More DD, Seidman JG, Smith JA, Struhl K (1997) Current protocols in molecular biology. Wiley-Interscience, New York

  7. Benzécri JP (1973) Analyse des données. Tome I: Analyse des correspondances. Tome II: La Classification. Dunod, Paris

    Google Scholar 

  8. Brígido C, Alexandre A, Laranjo M, Oliveira S (2007) Moderately acidophilic mesorhizobia isolated from chickpea. Lett Appl Microbiol 44:168–174

    Article  PubMed  Google Scholar 

  9. Brígido C, Alexandre A, Oliveira S Transcriptional analysis of major chaperone genes in salt-tolerant and salt-sensitive mesorhizobia. Microbiol Res. doi:10.1016/j.micres.2012.01.006

  10. Brockwell J, Pilka A, Holliday RA (1991) Soil pH is a major determinant of the numbers of naturally occurring Rhizobium meliloti in noncultivated soils in central New South Wales. Aust J Exp Agr 31:211–219

    Article  Google Scholar 

  11. Chen WX, Li GS, Qi YL, Wang ET, Yuan HL, Li JL (1991) Rhizobium huakuii sp. nov. isolated from the root-nodules of Astragalus sinicus. Int J Syst Bacteriol 41:275–280

    Article  Google Scholar 

  12. Chen WX, Wang E, Wang SY, Li YB, Chen XQ (1995) Characteristics of Rhizobium tianshanense sp. nov., a moderately and slowly growing root-nodule bacterium isolated from an arid saline environment in Xinjiang, Peoples Republic of China. Int J Syst Bacteriol 45:153–159

    Article  PubMed  CAS  Google Scholar 

  13. Chen WX, Wang ET, Kuykendall D (2005) Genus VI. Mesorhizobium. In: Bergey’s manual of systematic bacteriology, vol. 2 (The Proteobacteria). Springer, New York, pp. 403–408

  14. Correa OS, Barneix AJ (1997) Cellular mechanisms of pH tolerance in Rhizobium loti. World J Microbiol Biotechnol 13:153–157

    Article  CAS  Google Scholar 

  15. de Lucena DKC, Puehler A, Weidner S (2010) The role of sigma factor RpoH1 in the pH stress response of Sinorhizobium meliloti. BMC Microbiol 10(265):17

    Google Scholar 

  16. Dilworth MJ, Howieson JG, Reeve WG, Tiwari RP, Glenn AR (2001) Acid tolerance in legume root nodule bacteria and selecting for it. Aust J Exp Agr 41:435–446

    Article  CAS  Google Scholar 

  17. Fierer N, Jackson RB (2006) The diversity and biogeography of soil bacterial communities. Proc Natl Acad Sci USA 103:626–631

    Article  PubMed  CAS  Google Scholar 

  18. Foster JW (1991) Salmonella acid shock proteins are required for the adaptative acid tolerance response. J Bacteriol 173:6896–6902

    PubMed  CAS  Google Scholar 

  19. Frydman J (2001) Folding of newly translated proteins in vivo: the role of molecular chaperones. Annu Rev Biochem 70:603–647

    Article  PubMed  CAS  Google Scholar 

  20. Graham PH, Viteri SE, Mackie F, Vargas AT, Palacios A (1982) Variation in acid soil tolerance among strains of Rhizobium phaseoli. Field Crop Res 5(2):121–128

    Article  Google Scholar 

  21. Hartl FU (1996) Molecular chaperones in cellular protein folding. Nature 381:571–579

    Article  PubMed  CAS  Google Scholar 

  22. Hartl FU, Hayer-Hartl M (2002) Protein folding—molecular chaperones in the cytosol: from nascent chain to folded protein. Science 295:1852–1858

    Article  PubMed  CAS  Google Scholar 

  23. Hellweg C, Puhler A, Weidner S (2009) The time course of the transcriptomic response of Sinorhizobium meliloti 1021 following a shift to acidic pH. BMC Microbiol 9:37

    Article  PubMed  Google Scholar 

  24. Howieson JG, O’Hara GW, Carr SJ (2000) Changing roles for legumes in Mediterranean agriculture: developments from an Australian perspective. Field Crop Res 65:107–122

    Article  Google Scholar 

  25. Ibekwe AM, Angle JS, Chaney RL, vanBerkum P (1997) Enumeration and N-2 fixation potential of Rhizobium leguminosarum biovar trifolii grown in soil with varying pH values and heavy metal concentrations. Agric Ecosyst Environ 61:103–111

    Article  CAS  Google Scholar 

  26. Jakob U, Gaestel M, Engel K, Buchner J (1993) Small heat shock proteins are molecular chaperones. J Biol Chem 268:1517–1520

    PubMed  CAS  Google Scholar 

  27. Jarvis BDW, Pankhurst CE, Patel JJ (1982) Rhizobium loti, a new species of legume root nodule bacteria. Int J Syst Bacteriol 32:378–380

    Article  Google Scholar 

  28. Jarvis BDW, van Berkum P, Chen WX, Nour SM, Fernandez MP, Cleyet-Marel JC et al (1997) Transfer of Rhizobium loti, Rhizobium huakuii, Rhizobium ciceri, Rhizobium mediterraneum, and Rhizobium tianshanense to Mesorhizobium gen. nov. Int J Syst Bacteriol 47:895–898

    Article  Google Scholar 

  29. Kulkarni S, Nautiyal CS (1999) Characterization of high temperature-tolerant rhizobia isolated from Prosopis juliflora grown in alkaline soil. J Gen Appl Microbiol 45:213–220

    Article  PubMed  CAS  Google Scholar 

  30. Laguerre G, Courde L, Nouaim R, Lamy I, Revellin C, Breuil MC et al (2006) Response of rhizobial populations to moderate copper stress applied to an agricultural soil. Microb Ecol 52:426–435

    Article  PubMed  CAS  Google Scholar 

  31. Laranjo M, Oliveira S (2011) Tolerance of Mesorhizobium type strains to different environmental stresses. Anton Leeuw Int JG 99:651–662

    Article  CAS  Google Scholar 

  32. Lemos JA, Luzardo Y, Burne RA (2007) Physiologic effects of forced down-regulation of dnaK and groEL expression in Streptococcus mutans. J Bacteriol 189:1582–1588

    Article  PubMed  CAS  Google Scholar 

  33. Lemos JAC, Chen YYM, Burne RA (2001) Genetic and physiologic analysis of the groE operon and role of the HrcA repressor in stress gene regulation and acid tolerance in Streptococcus mutans. J Bacteriol 183:6074–6084

    Article  PubMed  CAS  Google Scholar 

  34. Li QQ, Wang ET, Zhang YZ, Zhang YM, Tian CF, Sui XH et al (2011) Diversity and biogeography of rhizobia isolated from root nodules of Glycine max grown in Hebei province, China. Microb Ecol 61:917–931

    Article  PubMed  Google Scholar 

  35. Marschner H (2006) Mineral nutrition of higher plants. Academic, London

    Google Scholar 

  36. Matsui R, Cvitkovitch D (2010) Acid tolerance mechanisms utilized by Streptococcus mutans. Future Microbiol 5:403–417

    Article  PubMed  CAS  Google Scholar 

  37. Nour SM, Cleyet-Marel J-C, Normand P, Fernandez MP (1995) Genomic heterogeneity of strains nodulating chickpeas (Cicer arietinum L.) and description of Rhizobium mediterraneum sp. nov. Int J Syst Bacteriol 45:640–648

    Article  PubMed  CAS  Google Scholar 

  38. Nour SM, Fernandez MP, Normand P, Cleyet-Marel J-C (1994) Rhizobium ciceri sp. nov., consisting of strains that nodulate chickpeas (Cicer arietinum L.). Int J Syst Bacteriol 44:511–522

    Article  PubMed  CAS  Google Scholar 

  39. Rao DLN, Giller KE, Yeo AR, Flowers TJ (2002) The effects of salinity and sodicity upon nodulation and nitrogen fixation in chickpea (Cicer arietinum). Ann Bot 89:563–570

    Article  PubMed  CAS  Google Scholar 

  40. Reeve WG, Brau L, Castelli J, Garau G, Sohlenkamp C, Geiger O et al (2006) The Sinorhizobium medicae WSM419 IpiA gene is transcriptionally activated by FsrR and required to enhance survival in lethal acid conditions. Microbiology 152:3049–3059

    Article  PubMed  CAS  Google Scholar 

  41. Rodrigues C, Laranjo M, Oliveira S (2006) Effect of heat and pH stress in the growth of chickpea mesorhizobia. Curr Microbiol 53:1–7

    Article  PubMed  CAS  Google Scholar 

  42. Ruiz-Díez B, Fajardo S, Puertas-Mejia MA, Felipe MD, Fernandez-Pascual M (2009) Stress tolerance, genetic analysis and symbiotic properties of root-nodulating bacteria isolated from Mediterranean leguminous shrubs in Central Spain. Arch Microbiol 191:35–46

    Article  PubMed  Google Scholar 

  43. Sabate R, de Groot NS, Ventura S (2010) Protein folding and aggregation in bacteria. Cell Mol Life Sci 67:2695–2715

    Article  PubMed  CAS  Google Scholar 

  44. Siddique KHM, Loss SP, Regan KL, Jettner RL (1999) Adaptation and seed yield of cool season grain legumes in Mediterranean environments of south-western Australia. Aust J Agr Res 50:375–387

    Article  Google Scholar 

  45. Torrent J, Barberis E, Gil-Sotres F (2007) Agriculture as a source of phosphorus for eutrophication in southern Europe. Soil Use Manage 23:25–35

    Article  Google Scholar 

  46. Vincent JM (1970) A manual for the practical study of root-nodule bacteria, no. 15. IBP Handbook. Blackwell, Oxford

    Google Scholar 

  47. Wang ET, van Berkum P, Sui XH, Beyene D, Chen WX, Martinez-Romero E (1999) Diversity of Rhizobia associated with Amorpha fruticosa isolated from Chinese soils and description of Mesorhizobium amorphae sp nov. Int J Syst Bacteriol 49:51–65

    Article  PubMed  Google Scholar 

  48. Wei Y, Zeng X, Yuan Y, Jiang H, Zheng Y, Tan Y et al (2011) DNA microarray analysis of acid-responsive genes of Streptococcus suis serotype 2. Ann Microbiol 61:505–510

    Article  CAS  Google Scholar 

  49. Zmijewski MA, Kwiatkowska JM, Lipinska B (2004) Complementation studies of the DnaK–DnaJ–GrpE chaperone machineries from Vibrio harveyi and Escherichia coli, both in vivo and in vitro. Arch Microbiol 182:436–449

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work has received funding from FCT (Fundação para a Ciência e a Tecnologia) and co-financed by EU-FEDER (PTDC/BIO/80932/2006). C. Brígido acknowledges a PhD fellowship (SFRH/BD/30680/2006) from FCT. We thank G. Mariano for technical assistance.

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  1. Laboratório de Microbiologia do Solo, ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Apartado 94, 7002-554, Évora, Portugal

    Clarisse Brígido & Solange Oliveira

  2. Departamento de Biologia, Universidade de Évora, Apartado 94, 7002-554, Évora, Portugal

    Solange Oliveira

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  1. Clarisse Brígido
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  2. Solange Oliveira
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Correspondence to Solange Oliveira.

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Brígido, C., Oliveira, S. Most Acid-Tolerant Chickpea Mesorhizobia Show Induction of Major Chaperone Genes upon Acid Shock. Microb Ecol 65, 145–153 (2013). https://doi.org/10.1007/s00248-012-0098-7

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