Plant and Soil

, Volume 298, Issue 1–2, pp 273–284 | Cite as

Mycorrhiza and soil bacteria influence extractable iron and manganese in soil and uptake by soybean

  • M. A. NogueiraEmail author
  • U. Nehls
  • R. Hampp
  • K. Poralla
  • E. J. B. N. Cardoso
Regular Article


Excess manganese (Mn) in soil is toxic to crops, but in some situations arbuscular mycorrhizal fungi (AMF) alleviate the toxic effects of Mn. Besides the increased phosphorus (P) uptake, mycorrhiza may affect the balance between Mn-reducing and Mn-oxidizing microorganisms in the mycorrhizosphere and affect the level of extractable Mn in soil. The aim of this work was to compare mycorrhizal and non-mycorrhizal plants that received extra P in relation to alleviation of Mn toxicity and the balance between Mn-oxidizing and Mn-reducing bacteria in the mycorrhizosphere. A clayey soil containing 508 mg kg−1 of extractable Mn was fertilized with 30 mg kg−1 (P1) or 45 mg kg−1 (P2) of soluble P. Soybean (Glycine max L. Merrill, cv. IAC 8-2) plants at P1 level were non-inoculated (CP1) or inoculated with either Glomus etunicatum (GeP1) or G. macrocarpum (GmP1), while plants at P2 level were left non-inoculated (CP2). Plants were grown in a greenhouse and harvested after 80 days. In the mycorrhizosphere of the GmP1 and GeP1 plants a shift from Mn-oxidizing to Mn-reducing bacteria coincided with higher soil extractability of Mn and Fe. However, the occurrence of Mn-oxidizing/reducing bacteria in the (mycor)rhizosphere was unrelated to Mn toxicity in plants. Using 16S rDNA sequence homologies, the Mn-reducing isolates were consistent with the genus Streptomyces. The Mn-oxidizers were homologous with the genera Arthrobacter, Variovorax and Ralstonia. While CP1 plants showed Mn toxicity throughout the whole growth period, CP2 plants never did, in spite of having Fe and Mn shoot concentrations as high as in CP1 plants. Mycorrhizal plants showed Mn toxicity symptoms early in the growth period that were no longer visible in later growth stages. The shoot P concentration was almost twice as high in mycorrhizal plants compared with CP1 and CP2 plants. The shoot Mn and Fe concentrations and contents were lower in GmP1 and GeP1 plants compared with the CP2 treatment, even though levels of extractable metals increased in the soil when plants were mycorrhizal. This suggests that mycorrhiza protected its host plant from excessive uptake of Mn and Fe. In addition, higher tissue P concentrations may have facilitated internal detoxification of Mn in mycorrhizal plants. The exact mechanisms acting on alleviation of Mn toxicity in mycorrhizal plants should be further investigated.


Fe Metal Mn Oxidation Reduction Toxicity 



arbuscular mycorrhizal fungi


colony forming units


non-mycorrhizal control at P level 1 (30 mg kg−1)


non-mycorrhizal control at P level 2 (45 mg kg−1)


Glomus macrocarpum at P level 1


Glomus etunicatum at P level 1


polymerase chain reaction



We thank Fundação de Apoio à Pesquisa do Estado de São Paulo and Coordenação de Aperfeiçoamento Pessoal de Nível Superior/Deutscher Akademischer Austauschdienst/Programa Brasil-Alemanha for financial support. We acknowledge Denise L.C. Mescolotti, Luís F. Baldesin, and Rafaela F. Neroni for their help in setting up and conducting the experiments of this study. Thanks are also due to Dr. A. Leyva who provided English language editing of the manuscript, Dr. Volker Roemheld (section editor), and the two anonymous referees. M.A. Nogueira and E.J.B.N. Cardoso are CNPq scholars.


  1. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  2. Andrade G, Mihara KL, Linderman RG, Bethlenfalvay GJ (1997) Bacteria from rhizosphere and hyphosphere soils of different arbuscular–mycorrhizal fungi. Plant Soil 192:71–79CrossRefGoogle Scholar
  3. Andrade G, Linderman RG, Bethlenfalvay GJ (1998) Bacterial associations with mycorrhizosphere and hyphosphere of the arbuscular mycorrhizal fungus Glomus mosseae. Plant Soil 202:79–87CrossRefGoogle Scholar
  4. Bromfield SM (1974) Bacterial oxidation of manganous ions as affected by organic substrate concentration and composition. Soil Biol Biochem 6:383–392CrossRefGoogle Scholar
  5. Bromfield SM, Skerman VBD (1950) Biological oxidation of manganese in soils. Soil Sci 69:337–347CrossRefGoogle Scholar
  6. Cardoso EJBN (1996) Interaction of mycorrhiza, phosphate and manganese in soybean. In: Mycorrhizas in integrated systems: from genes to plant development. (ed) C Azcón-Aguilar and J M Barea. Proceedings of the IV European Symposium on Mycorrhizas, Granada, July 1994. Luxemburg, European Commission Report, pp 304–306Google Scholar
  7. Corstjens PLAM, de Vrind JPM, Wesbroek P, de Vrind-de Jong EW (1992) Enzymatic iron oxidation by Leptotrhix discophora: identification of an iron-oxidizing protein. Appl Environ Microb 58:450–454Google Scholar
  8. Foy CD (1984) Physiological effects of hydrogen, aluminium, and manganese toxicities in acid soils. In: Adams F (ed) Soil Acidity and Liming. pp 57–97. American Society of Agronomy, MadisonGoogle Scholar
  9. Ghiorse WC (1984) Biology of iron- and manganese-depositing bacteria. Annu Rev Microbiol 38:515–550PubMedGoogle Scholar
  10. Gilbert ES, Crowley DE (1998) Repeated application of carvone-induced bacteria to enhance biodegradation of polychlorinated biphenyls in soil. Appl Microbiol Biotechnol 50:489–494PubMedCrossRefGoogle Scholar
  11. Giovannetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular arbuscular mycorrhizal infection in roots. New Phytol 84:489–500CrossRefGoogle Scholar
  12. Hall JL (2002) Cellular mechanisms for heavy metal detoxification and tolerance. J Exp Bot 53:1–11PubMedCrossRefGoogle Scholar
  13. Hayatsu M, Hirano M, Nagata T (1999) Involvement of two plasmids in the degradation of carbaryl by Arthrobacter sp. strain RC100. Appl Environ Microb 65:1015–1019Google Scholar
  14. Huber DM, Graham RD (1992) Techniques for studying nutrient disease interactions. In: Singleton LL, Mihail JD, Rush CM (ed) Methods for Research on Soilborne Phytopathogenic Fungi. pp 204–214. APS Press, Saint PaulGoogle Scholar
  15. Joner EJ, Briones R, Leyval C (2000) Metal-binding capacity of arbuscular mycorrhizal mycelium. Plant Soil 226:227–234CrossRefGoogle Scholar
  16. Kothari SK, Marschner H, Römheld V (1991) Effect of a vesicular–arbuscular mycorrhizal fungus and rhizosphere micro-organisms on manganese reduction in the rhizosphere and manganese concentrations in maize (Zea mays L.). New Phytol 117:649–655CrossRefGoogle Scholar
  17. Marschner H (1988) Mechanism of manganese acquisition by roots from soils. In: Graham RD Hannan RJ, Uren NC (ed) Manganese in Soil and Plants. pp 191–204. Kluwer, BostonGoogle Scholar
  18. Marschner H (1995) Mineral Nutrition of Higher Plants. Academic, London, 889 ppGoogle Scholar
  19. Marschner P, Ascher JS, Graham RD (1991) Effect of manganese-reducing rhizosphere bacteria on the growth of Gaeumannomyces graminis var. tritici and on manganese uptake by wheat (Triticum aestivum L.). Biol Fert Soils 12:33–38CrossRefGoogle Scholar
  20. Mehlich A (1953) Determination of P, Ca, Mg, K, Na, and \( {\text{NH}}^{ + }_{4} \). North Carolina Soil Test Division (Mimeo 1953). North Carolina Dept. of Agric., Raleigh, NCGoogle Scholar
  21. Nealson KH, Tebo BM, Rosson RA (1988) Occurrence and mechanisms of microbial oxidation of manganese. Adv Appl Microbiol 33:279–318CrossRefGoogle Scholar
  22. Nogueira MA, Cardoso EJBN (2002) Microbial interactions on manganese availability and uptake by soybean. Pesqui. Agropecu Bras 37:1605–1612Google Scholar
  23. Nogueira MA, Cardoso EJBN, Hampp R (2002) Manganese toxicity and callose deposition in leaves are attenuated in mycorrhizal soybean. Plant Soil 246:1–10CrossRefGoogle Scholar
  24. Nogueira MA, Magalhães GC, Cardoso EJBN (2004) Manganese toxicity in mycorrhizal and phosphorus-fertilized soybean plants. J Plant Nutr 27:141–156CrossRefGoogle Scholar
  25. Okazaki M, Sugita T, Shimizu M, Ohode Y, Iwamoto K, de Vrind-de Jong de Vrind JPM, Corstjens PLAM (1997) Partial purification and characterization of manganese-oxidizing factors of Pseudomonas fluorescens GB-1. Appl Environ Microb 63:4793–4799Google Scholar
  26. Parales RE, Ditty JL, Harwood CS (2000) Toluene-degrading bacteria are chemotactic towards the environmental pollutants benzene, toluene, and trichloroethylene. Appl Environ Microb 66:4098–4104CrossRefGoogle Scholar
  27. Phillips JM, Hayman AS (1970) Improved procedures for clearing roots and staining parasitic and vesicular–arbuscular mycorrhizal fungi for assessment of infection. Trans Brit Mycol Soc 55:158–161CrossRefGoogle Scholar
  28. Posta K, Marschner H, Römheld V (1994) Manganese reduction in the rhizosphere of mycorrhizal and nonmycorrhizal maize. Mycorrhiza 5:119–124CrossRefGoogle Scholar
  29. Ramos CP, Foster G, Collins MD(1997) Phylogenetic analysis of the genus Actinomyces based on 16S rRNA gene sequences: Description of an Arcanobacterium phocae sp. nov., Arcanobacterium bernardiae comb. nov., and Arcanobacterium pyogenes comb. nov. Int J Syst Bacteriol 47:46–53PubMedCrossRefGoogle Scholar
  30. Ridge EH, Rovira AD (1971) Phosphatase activity of intact young wheat roots under sterile and non-sterile conditions. New Phytol 70:1017–1026CrossRefGoogle Scholar
  31. Rogalla H, Römheld V (2002) Role of leaf apoplast in silicon-mediated manganese tolerance of Cucumis sativus L. Plant Cell Environ 25:549–555CrossRefGoogle Scholar
  32. Sarruge JR, Haag HP (1974) Análises Químicas em Plantas. Escola Superior de Agricultura Luiz de Queiroz, Piracicaba, 56 ppGoogle Scholar
  33. SAS (1991) The Statistical Analysis System. Procedure Guide for Personal Computers. Release 6.11. 5th ed. Cary, USA, 649 ppGoogle Scholar
  34. Schenk S, Decker K (1999) Horizontal gene transfer involved in the convergent evolution of the plasmid-encoded enantioselective 6-hydroxynicotine oxidases. J Mol Evol 48:178–186PubMedCrossRefGoogle Scholar
  35. Soil Survey Staff 1999 Soil Taxonomy: a Basic System of Soil Classification for Making and Interpreting Soil Surveys. USDA, Washington. 869 ppGoogle Scholar
  36. Sparks DL, Page AL, Helmke PA, Loeppert RH, Soltanpour PN, Tabatabai MA, Johnston CT, Summer ME (1996) Methods of Soil Analysis. Part 3: Chemical Methods. Soil Science Society of America, Madison, Wiscosin, p 1390Google Scholar
  37. Thompson JA, Huber DM, Guest CA, Schulze DG (2005) Fungal manganese oxidation in a reduced soil. Environ Microbiol 7:1480–1487PubMedCrossRefGoogle Scholar
  38. Yano K, Takaki M (2005) Mycorrhizal alleviation of acid soil stress in sweet potato (Ipomoea batatas). Soil Biol Biochem 37:1569–1572CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • M. A. Nogueira
    • 1
    Email author
  • U. Nehls
    • 2
  • R. Hampp
    • 2
  • K. Poralla
    • 3
  • E. J. B. N. Cardoso
    • 4
  1. 1.CCB/Department of MicrobiologyState University of Londrina, Lab. for Microbial EcologyLondrinaBrazil
  2. 2.University of Tübingen, Institute of Botany/Physiological Ecology of PlantsTübingenGermany
  3. 3.Department of BiologyUniversity of TübingenTübingenGermany
  4. 4.Department of Soil ScienceUniversity of São PauloPiracicabaBrazil

Personalised recommendations