Skip to main content
Log in

Azospirillum strains use phenolic compounds as intermediates for electron transfer under oxygen-limiting conditions

  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

The effects of catechol, vanillic, caffeic (CAF), 2-hydroxyphenylacetic, 4-hydroxy- and 3,4-dihydroxybenzoic (3,4-DHBA) acids on the growth of a common rice rhizosphere inhabitant, Azospirillum lipoferum were studied. Two strains of this nonfermenting nitrogen-fixing bacterium were used: a motile strain (4B), and a nonmotile strain (4T). Under atmospheric conditions (pO2 = 21 kPa), the growth of strain 4T was inhibited by catechol (0.1 mm) only. None of these compounds affected the growth of strain 413. Under 5 kPa O2, no effect was observed on strain 413, whereas three of the six tested phenolics stimulated the growth of strain 4T; maximum effects were observed for 3,4-DHBA and CAF. As revealed by TLC and HPLC, under low oxygen, more new lipophilic compounds were formed from CAF by strain 4T, differing from CAF autooxydation products and from the products obtained under 21 kPa O2. It was hypothesized that strain 4T had the ability to use an oxidized derivative of CAF as a terminal electron acceptor. This hypothesis was tested in experiments under nitrogen-fixing conditions, in the absence of oxygen, and in the presence of N2O as a reoxidizing agent for CAF. Acetylene was used both as a substrate to measure nitrogenase activity (ARA) and to inhibit the biological transfer of electrons to N2O. The addition of CAF in the presence of N2O had the same effect on ARA rates as an addition of oxygen. It is concluded that the strain 4T of Azospirillum lipoferum is able to sustain some of its activities (e.g., N2 fixation) using phenolics as alternative electron acceptors under low oxygen conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Atlas RM, Bartha R (1981) Microbial ecology: fundamentals and applications. Addison-Wesley Pub., Reading, Mass.

    Google Scholar 

  2. Bally R, Thomas-Bauzon D, Heulin T, Balandreau J, Richard C, De Ley J (1983) Determination of the most frequent N2-fixing bacteria in a rice rhizosphere. Can J Microbiol 29:881–887

    Google Scholar 

  3. Barkovskii AL (1993) Biodestructive potential of symbiotic and of associative microorganisms to aromatic and haloaromatic compounds. In: Edyvean RG (ed) Effluent treatment and waste minimisation. (Chem Symp. ser., 132) Chameleon Press, London, pp 157–169

    Google Scholar 

  4. Barkovskii AL, Shub GM (1986) An Acinetobacter calcoaceticus strain utilizing various aromatic compounds and carrying a plasmid for resorcinol degradation. Mikrobiologija 55:237–240

    Google Scholar 

  5. Bassam BJ, Djordjevic MA, Redmond JW, Batley M, Rolfe BG (1988) Identification of a nodD-dependent locus in the Rhizobium strain NG234 activated by phenolic factor secreted by soybeans and other legumes. Mol Plant-Microbe Interact 1:161–168

    Google Scholar 

  6. Blum U, Wentworth TR, Klein K, Worsham AD, King LD, Geric TM, Lyu SW (1991) Phenolic acid content of soil from wheat no-till, wheat conventional-till, and fallow conventional-till soybean cropping system. J Chem Ecol 17:1045–1068

    Google Scholar 

  7. Burns RC, Hardy RWF (1975) Nitrogen fixation in bacteria and higher plants. Springer-Verlag, Berlin

    Google Scholar 

  8. Crane FL, Barr R (1985) Chemical structure and properties of coenzyme Q and related compounds. In: Lenaz G (ed) Coenzyme Q. Biochemistry, bioenergetics, and clinical applications of ubiquinone. Wiley & Sons, New York, pp 1–39

    Google Scholar 

  9. Dagley S (1967) The microbial metabolism of phenolics. In: McLaren AD, Peterson GH (eds) Soil biochemistry. Marcel Dekker, Inc., New York, pp 287–317

    Google Scholar 

  10. Deiana S, Gessa C, Manunza B, Marchetti M, Usai M (1992) Mechanism and stoichiometry of the redox reaction between iron (III) and caffeic acid. Plant Soil 145:287–294

    Article  CAS  PubMed  Google Scholar 

  11. Einarsdottir GH, Stankovich MT, Tu S-C (1988) Studies of electron-transfer properties of salicylate hydroxylase from Pseudomonas cepacia and effects of salicylate and benzoate binding. Biochemistry 27:3277–3285

    Google Scholar 

  12. Flaig W, Beutelspacher H, Rietz E (1975) Chemical composition and physical properties of humic substances. In: Gieseking JE (ed) Soil components: v.l. Organic components. Springer-Verlag, Berlin, pp 1–211

    Google Scholar 

  13. Flessa A, Fischer WR (1992) Plant-induced changes in the redox potential of rice rhizospheres. Plant Soil 143:55–60

    CAS  Google Scholar 

  14. Gajendiran N, Mahadevan A (1990) Plasmid-born catechol dissimilation in Rhizobium sp. FEMS Microbiol Ecol 73:125–130

    Google Scholar 

  15. Gasser F, Biville F, Turline G (1991) Un nouveau cofacteur d'oxydoreduction, la pyrroloquinoline quinone. Ann Inst Pasteur/Actualités 2:139–149

    Google Scholar 

  16. Givaudan A, Effosse A, Bally R (1991) Melanin production by Azospirillum lipoferum strains. In: Polsinelli M, Materassi R, Vincenzini M (eds) Nitogen fixation. (Developments in plant and soil sciences) Kluwer Academic, Dordrecht, pp 311–312

    Google Scholar 

  17. Givaudan A, Effosse A, Faure D, Potier P, Bouillant ML, Bally R (1993) Polyphenol oxidase in Azospirillum lipoferum isolated from rice rhizosphere: evidence of a laccase activity in non-motile strains of Azospirillum lipoferum. FEMS Microbiol Let 108:205–210

    Google Scholar 

  18. Haider K, Martin JP (1975) Decomposition of specifically carbon-14 labelled benzoic acid and cinnamic acid derivatives in soil. Soil Sci Soc Am Proc 39:657–662

    Google Scholar 

  19. Hardwood CS, Rivelli M, Orston LN (1984) Aromatic acids are chemoattractants for Pseudomonas putida. J Bacteriol 169:622–628

    Google Scholar 

  20. Harris RF (1982) Energetics of nitrogen transformations. In: Stevenson FJ (ed) Nitrogen in agricultural soils. (Agronomy series 22) ASA, CSA, and SSSA Pub, Madison, Wisconsin, pp 833–890

    Google Scholar 

  21. Hartley RD, Whitehead DC (1985) Phenolic acids in soils and their influence on plant growth and soil microbial processes. In: Vaughan D, Malcolm RE (eds) Soil organic matter and biological activity. Martinus Nijhoff/DR W. Junk, Dordrecht, Germany, pp 109–149

    Google Scholar 

  22. Hartmann A, Fusseder A, Klingmuller W (1983) Mutants of Azospirillum affected in nitrogen fixation and auxin production. In: Klingmüller W (ed) Azospirillum. II. Genetics, physiology, ecology. (Experientia Suppl. 48) Birhäuser Verlag, Basel, pp 78–88

    Google Scholar 

  23. Havelka UD, Boyle MG, Hardy RWF (1982) Biological nitrogen fixation. In: Stevenson FJ (ed) Nitrogen in agricultural soils. (Agronomy series 22) ASA, CSA, and SSSA Pub, Madison, Wisconsin, pp 365–422

    Google Scholar 

  24. Heulin T, Weinhard P, Balandreau J (1983) Motility changes in Azospirillum lipoferum. In: Klingmuller W (ed) Azospirillum. II. Genetics, physiology, ecology. (Experientia Suppl 48) Birkhäuser Verlag, Basel, pp 89–94

    Google Scholar 

  25. Kape R, Parniske M, Werner D (1991) Chemotaxis and nod gene activity of Bradyrhizobium japonicum in response to hydroxycinnamic acids and isoflavonoids. Appl Environ Microbiol 57:316–319

    Google Scholar 

  26. Karasevitch Yu N (1982) The foundation of selection for microorganisms utilizing synthetic organic compounds. Mir, Moscow

    Google Scholar 

  27. Korzhenevitch VI, Ignatov OV, Mironov AD, Krivopalov VV, Barkovskii AL (1991) Activity of bacterial strains destructors of aromatic compounds entrapped in agar gel beads. Prikladnaja Biokhim Mikrobiol 27:365–369

    Google Scholar 

  28. Krotzky A, Berggold R, Jaeger D, Dart PJ, Werner D (1983) Enhancement of aerobic nitrogenase activity (acetylene reduction assay) by phenol in soils and the rhizosphere of cereals. Z Pflanzenernaeher Bodenkd 146:634–642

    Google Scholar 

  29. Kuiters AT, Dennemann CAJ (1987) Water soluble phenolic substances in soils under several coniferous and deciduous tree species. Soil Biol Biochem 19:765–769

    Google Scholar 

  30. Kumada K (1987) Chemistry of soil organic matter. Elsevier, Amsterdam

    Google Scholar 

  31. Lehmann RG, Chang HH (1988) Reactivity of phenolic acids in soil and formation of oxidation products. Soil Sci Soc Am J 52:1304–1309

    Google Scholar 

  32. Lehmann RG, Cheng HH, Harsh JB (1987) Oxidation of phenolic acids by soil iron and manganese oxides. Soil Sci Soc Am J 51:352–356

    Google Scholar 

  33. Le Strange KK, Bender GL, Djordjevic MA, Rolfe BG, Redmond JW (1990) The Rhizobium strain NGR234 nodD1 gene product responds to activation by the simple phenolic compounds vanillin and isovanillin present in wheat seedling extracts. Mol Plant-Microbe Interact 3–4:214–220

    Google Scholar 

  34. Lynch JM, Whipps JM (1990) Substrate flow in the rhizosphere. Plant Soil 129:1–10

    Google Scholar 

  35. Lynn DG, Chang M (1990) Phenolic signals in cohabitation: implications for plant development. Annu Rev Plant Physiol Plant Mol Biol 41:497–526

    Google Scholar 

  36. Pflug W, Ziechmann W (1981) Inhibition of malate dehydrogenase by humic acids. Soil Biol Biochem 13:239–299

    Google Scholar 

  37. Schnitzer M (1978) Humic substances: chemistry and reactions. In: Schnitzer M, Khan SU (eds) Soil organic matter. Elsevier, Amsterdam, pp 1–64

    Google Scholar 

  38. Spaling GP, Ord BG, Vaughan D (1981) Changes in microbial biomass and activity in soils amended with phenolic compounds. Soil Biol Biochem 13:455–460

    Google Scholar 

  39. Swallow AJ (1982) Physical chemistry of semiquinones. In: Trumpower BL (ed) Function of quinones in energy conserving systems. Academic Press, London, pp 59–73

    Google Scholar 

  40. Thomas-Bauzon D, Weinhard P, Villecourt P, Balandreau J (1982) The spermosphere model. 1. Its use in growing, counting, and isolating nitrogen-fixing bacteria from the rizosphere of rice. Can J Microbiol 28:922–928

    Google Scholar 

  41. Ueckert J, Hurek T, Fendrik I, Niemann EG (1990) Radial gas diffusion from roots of rice (Oryza sativa L.) and Kallar grass (Leptochloa fuska L. Kunth. ), and effect of inoculation with Azospirillum brasifense Cd. Plant Soil 122:59–65

    Google Scholar 

  42. Vançura V, Kunc F (1988) Soil microbial associations: control of structures and functions. Elsevier, Amsterdam

    Google Scholar 

  43. von Ziechmann W (1972) deÜber die elektronen-Donator-und-Acceptor-Eingenschaften von Huminstoffen. Geoderma 8:111–131

    Google Scholar 

  44. von Ziechmann W (1977) MolekülKomplexe bei Huminstoffen durch e-Donator- und e-AcceptorStrukturen. Z Pflanzenernaehr Bodenkd 140:133–150

    Google Scholar 

  45. Werner D, Krotzky A, Berggold R, Thierfelder H, Preiss M (1982) Enhancement of specific nitrogenase activity in Azospirillum brasilense and Klebsiella pneumoniae, inhibition in Rhizobium japonicum under air by phenol. Arch Microbiol 132:51–56

    Google Scholar 

  46. Whitehead DC, Dibb H, Hartley RD (1983) Bound phenolic compounds in water extracts of soils, plant roots, and leaf litter. Soil Biol Biochem 15:133–136

    Google Scholar 

  47. Yoshinari T, Hynes R, Knowles R (1977) Acetylene inhibition of nitrous oxide reduction and measurement of denitrification and nitrogen fixation in soil. Soil Biol Biochem 9:177–183

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Barkovskii, A., Bouillant, M.L., Monrozier, L.J. et al. Azospirillum strains use phenolic compounds as intermediates for electron transfer under oxygen-limiting conditions. Microb Ecol 29, 99–114 (1995). https://doi.org/10.1007/BF00217426

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00217426

Keywords

Navigation