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Naturwissenschaften

, Volume 94, Issue 10, pp 821–828 | Cite as

Non-specific association between filamentous bacteria and fungus-growing ants

  • Christian Kost
  • Tanja Lakatos
  • Ingo Böttcher
  • Wolf-Rüdiger Arendholz
  • Matthias Redenbach
  • Rainer Wirth
Original Paper

Abstract

Fungus-growing ants and their fungal cultivar form a highly evolved mutualism that is negatively affected by the specialized parasitic fungus Escovopsis. Filamentous Pseudonocardia bacteria occurring on the cuticle of attine ants have been proposed to form a mutualistic interaction with these ants in which they are vertically transmitted (i.e. from parent to offspring colonies). Given a strictly vertical transmission of Pseudonocardia, the evolutionary theory predicts a reduced genetic variability of symbionts among ant lineages. The aim of this study was to verify whether actinomycetes, which occur on Acromyrmex octospinosus leaf-cutting ants, meet this expectation by comparing their genotypic variability with restriction fragment length polymorphisms. Multiple actinomycete strains could be isolated from both individual ant workers and colonies (one to seven strains per colony). The colony specificity of actinomycete communities was high: Only 15% of all strains were isolated from more than one colony, and just 5% were present in both populations investigated. Partial sequencing of 16S ribosomal deoxyribonucleic acid of two of the isolated strains assigned both of them to the genus Streptomyces. Actinomycetes could also be isolated from workers of the two non-attine ant species Myrmica rugulosa and Lasius flavus. Sixty-two percent of the strains derived from attine ants and 80% of the strains isolated from non-attine ants inhibited the growth of Escovopsis. Our data suggest that the association between attine ants and their actinomycete symbionts is less specific then previously thought. Soil-dwelling actinomycetes may have been dynamically recruited from the environment (horizontal transmission), probably reflecting an adaptation to a diverse community of microbial pathogens.

Keywords

Actinomycetes Leaf-cutting ants Acromyrmex octospinosus Escovopsis Streptomyces Tripartite mutualism 

Notes

Acknowledgements

We thank Silvia Schmidt and Pascal Petronelli for invaluable help during fieldwork and Stefanie Bohnert for support in the laboratory. Constructive suggestions on the study and the manuscript by Anne Behrend, Hubert Herz, Michael Lakatos, Sandra Patiño, Silvia Schmidt, Dieter Spiteller and four anonymous referees are gratefully acknowledged. The Smithsonian Tropical Research Institute of the Republic of Panama assisted with the research and granted collecting permits. This work was supported by the European Large Scale Facility (‘Silvolab,’ French Guiana, no. DG/FJ/033/98) to Rainer Wirth. All experiments complied with the current laws in Germany, French Guiana and Panama.

References

  1. Bass M, Cherrett JM (1994) The role of leaf-cutting ant workers (Hymenoptera: Formicidae) in fungus garden maintenance. Ecol Entomol 19:215–220Google Scholar
  2. Bidet P, Lalande V, Salauze B, Burghoffer B, Avesani V, Delmee M, Rossier A, Barbut F, Petit JC (2000) Comparison of PCR-ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for typing Clostridium difficile. J Clin Microbiol 38:2484–2487PubMedGoogle Scholar
  3. Bot ANM, Rehner SA, Boomsma JJ (2001) Partial incompatibility between ants and symbiotic fungi in two sympatric species of Acromyrmex leaf-cutting ants. Evolution 55:1980–1991PubMedGoogle Scholar
  4. Boucher DH (1988) The biology of mutualism: ecology and evolution. Oxford Univ. Press, New YorkGoogle Scholar
  5. Cafaro MJ, Currie CR (2005) Phylogenetic analysis of mutualistic filamentous bacteria associated with fungus-growing ants. Can J Microbiol 51:441–446PubMedCrossRefGoogle Scholar
  6. Chapela IH, Rehner SA, Schultz TR, Mueller UG (1994) Evolutionary history of the symbiosis between fungus-growing ants and their fungi. Science 266:1691–1697CrossRefPubMedGoogle Scholar
  7. Currie CR, Mueller UG, Malloch D (1999a) The agricultural pathology of ant fungus gardens. Proc Natl Acad Sci USA 96:7998–8002CrossRefGoogle Scholar
  8. Currie CR, Scott JA, Summerbell RC, Malloch D (1999b) Fungus-growing ants use antibiotic-producing bacteria to control garden parasites. Nature 398:701–704CrossRefGoogle Scholar
  9. Currie CR, Scott JA, Summerbell RC, Malloch D (2003) Corrigendum: fungus-growing ants use antibiotic-producing bacteria to control garden parasites. Nature 423:461–461CrossRefGoogle Scholar
  10. Currie CR, Poulsen M, Mendenhall J, Boomsma JJ, Billen J (2006) Coevolved crypts and exocrine glands support mutualistic bacteria in fungus-growing ants. Science 311:81–83PubMedCrossRefGoogle Scholar
  11. Douglas AE (1994) Symbiotic interactions. Oxford Univ. Press, New YorkGoogle Scholar
  12. Douglas AE (1995) The ecology of symbiotic microorganisms. Adv Ecol Res 26:69–103Google Scholar
  13. Farish DJ (1972) Evolutionary implications of qualitative variation in grooming behavior of Hymenoptera (Insecta). Anim Behav 20:662–676PubMedCrossRefGoogle Scholar
  14. Fernandez C, Szabó IM (1978) Composition and properties of the actinomycete flora in a ferralitic tropical soil (oxisol)–sugar cane ecological system. Zentralbl Bakteriol Naturwiss 133:34–44PubMedGoogle Scholar
  15. Frank SA (1997) Models of symbiosis. Am Nat 150:S80–S99CrossRefGoogle Scholar
  16. Frank SA (2003) Perspective: repression of competition and the evolution of cooperation. Evolution 57:693–705PubMedGoogle Scholar
  17. Goodfellow M, Cross T (1984) The biology of actinomycetes. Academic, LondonGoogle Scholar
  18. Gotelli NJ, Colwell RK (2001) Quantifying biodiversity: procedures and pitfalls in the measurement and comparison of species richness. Ecol Lett 4:379–391CrossRefGoogle Scholar
  19. Hamilton WD, May RM (1977) Dispersal in stable habitats. Nature 269:578–581CrossRefGoogle Scholar
  20. Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of streptomyces. A laboratory manual. John Innes Foundation, NorwichGoogle Scholar
  21. Hughes WOH, Thomsen L, Eilenberg J, Boomsma JJ (2004) Diversity of entomopathogenic fungi near leaf-cutting ant nests in a neotropical forest, with particular reference to Metarhizium anisopliae var. anisopliae. J Invertebr Pathol 85:46–53PubMedCrossRefGoogle Scholar
  22. Kieser T, Mervyn JB, Buttner MJ, Chater KF, Hopwood DA (2000) General introduction to actinomycete biology. In: Kieser T, Mervyn JB, Buttner MJ, Chater KF, Hopwood DA (eds) Practical Streptomyces genetics. The John Innes Foundation, Norwich, pp 1–41Google Scholar
  23. Lee JY, Hwang BK (2002) Diversity of antifungal actinomycetes in various vegetative soils of Korea. Can J Microbiol 48:407–417PubMedCrossRefGoogle Scholar
  24. Maniatis T, Fritsch EF, Sambrook J (1989) Molecular Cloning—a laboratory manual. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  25. Martin MM (1970) The biochemical basis of the fungus–attine ant symbiosis. Science 169:16–20PubMedCrossRefGoogle Scholar
  26. Maschwitz U (1974) Comparative studies on the function of the metapleural gland in ants. Oecologia 16:303–310CrossRefGoogle Scholar
  27. Mühlenberg M (1993) Freilandökologie. Quelle and Meyer, HeidelbergGoogle Scholar
  28. North RD, Jackson CW, Howse PE (1997) Evolutionary aspects of ant-fungus interactions in leaf-cutting ants. Trends Ecol Evol 12:386–389CrossRefGoogle Scholar
  29. Pernodet JL, Boccard F, Alegre MT, Gagnat J, Guerineau M (1989) Organization and nucleotide sequence analysis of a ribosomal RNO gene cluster from Streptomyces ambofaciens. Gene 79:33–46PubMedCrossRefGoogle Scholar
  30. Poulsen M, Boomsma JJ (2005) Mutualistic fungi control crop diversity in fungus-growing ants. Science 307:741–744PubMedCrossRefGoogle Scholar
  31. Poulsen M, Currie CR (2006) Complexity of insect–fungal associations: exploring the influence of microorganisms on attine ant–fungus symbiosis. In: Bourtzis K, Miller TA (eds) Insect symbiosis, vol. 2. CRC, Boca Raton, FL, pp 57–77Google Scholar
  32. Poulsen M, Bot ANM, Nielsen MG, Boomsma JJ (2002) Experimental evidence for the costs and hygienic significance of the antibiotic metapleural gland secretion in leaf-cutting ants. Behav Ecol Sociobiol 52:151–157CrossRefGoogle Scholar
  33. Poulsen M, Cafaro M, Boomsma JJ, Currie CR (2005) Specificity of the mutualistic association between actinomycete bacteria and two sympatric species of Acromyrmex leaf-cutting ants. Mol Ecol 14:3597–3604PubMedCrossRefGoogle Scholar
  34. Ranzoni FV (1968) Fungi isolated in culture from soils of the Sonoran desert. Mycologia 60:356–371PubMedCrossRefGoogle Scholar
  35. Redenbach M, Kieser HM, Denapaite D, Eichner A, Cullum J, Kinashi H, Hopwood DA (1996) A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol 21:77–96PubMedCrossRefGoogle Scholar
  36. Rodrigues CRR, Drozdowicz (1978) The occurrence of Actinomycetes in a Cerrado soil in Brazil. Rev Écol Biol Sol 15:459–473Google Scholar
  37. Rodrigues A, Pagnocca FC, Bueno OC, Pfenning LH, Bacci M (2005) Assessment of microfungi in fungus gardens free of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae). Sociobiology 46:329–334Google Scholar
  38. Schmid-Hempel P (1998) Parasites in social insects. Princeton Univ. Press, PrincetonGoogle Scholar
  39. Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray RG, Wood WA, Krieg NR (eds) Methods for general and molecular microbiology. American Society for Microbiology, Washington, DC, pp 611–654Google Scholar
  40. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The Clustal X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  41. von Ihering H (1898) Die Anlage neuer Colonien und Pilzgärten bei Atta sexdens. Zool Anz 21:238–245Google Scholar
  42. Waksman SA, Lechevalier HA (1962) The actinomycetes, vol 3. Antibiotics of actinomycetes. Williams and Wilkinson, BaltimoreGoogle Scholar
  43. Wang Y, Zhang ZS, Ruan JS, Wang YM, Ali SM (1999) Investigation of actinomycete diversity in the tropical rainforests of Singapore. J Ind Microbiol Biotechnol 23:178–187CrossRefGoogle Scholar
  44. Weber NA (1966) Fungus growing ants. Science 153:587–604PubMedCrossRefGoogle Scholar
  45. Wheeler WM (1910) Colonies of ants (Lasius neoniger Emery) infested with Laboulbenis formicarum Thaxter. Psyche 17:83–86CrossRefGoogle Scholar
  46. Wilkinson DM, Sherratt TN (2001) Horizontally acquired mutualisms, an unsolved problem in ecology? Oikos 92:377–384CrossRefGoogle Scholar
  47. Wirth R, Beyschlag W, Ryel R, Herz H, Hölldobler B (2003) The herbivory of leaf-cutting ants. A case study on Atta colombica in the tropical rainforest of Panama. Springer, BerlinGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Christian Kost
    • 1
    • 2
    • 4
  • Tanja Lakatos
    • 1
  • Ingo Böttcher
    • 2
    • 3
  • Wolf-Rüdiger Arendholz
    • 1
  • Matthias Redenbach
    • 2
  • Rainer Wirth
    • 1
  1. 1.Department of Plant Ecology and SystematicsTechnical University of KaiserslauternKaiserslauternGermany
  2. 2.Department of Genetics—Genome Research UnitUniversity of KaiserslauternKaiserslauternGermany
  3. 3.Department of DermatologyUniversity MainzMainzGermany
  4. 4.School of Biological SciencesUniversity of AucklandAucklandNew Zealand

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