The Laboratory Environment Affects the Volatiles of Fungus Gardens in the Colonies of Fungus-farming Ants

Authors

  • Diego Santana Assis 1- Depto. de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Univ. de São Paulo (USP), Ribeirão Preto-SP, Brazil https://orcid.org/0000-0002-8131-2617
  • Ted Schultz 2- Department of Entomology, National Museum of Natural History, Smithsonian Institution, Washington, DC, USA
  • Skylar Brodowski 2- Department of Entomology, National Museum of Natural History, Smithsonian Institution, Washington, DC, USA
  • G. Asher Newsome Museum Conservation Institute, Smithsonian Institution, 4210 Silver Hill Road, Suitland, MD, USA
  • Fabio Santos do Nascimento 1- Depto. de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Univ. de São Paulo (USP), Ribeirão Preto-SP, Brazil

DOI:

https://doi.org/10.13102/sociobiology.v72i1.9768

Keywords:

Attina, Volatiles, DEET, Naphthalene, Mutualism

Abstract

The ability to recognize nestmates is critical to the ecological success of social insects. Fungus-farming “attine” ants (Formicidae: Myrmicinae: Attini: Attina) can recognize their nestmates and symbiotic fungi via chemoreception. Although it has been shown that mutualistic fungi release volatile compounds that elicit responses in fungus-farming ants, the compounds and the sensory mechanisms involved remain little studied. Here, we characterize compounds found in attine fungus gardens and explore the correlations between those compounds, fungal substrates, and the laboratory environment. We also characterize ant cuticular hydrocarbons from Atta cephalotes colonies of the same species maintained in the same laboratory conditions for two or more years. Using gas chromatography associated with mass spectrometry, we verified that both substrate (i.e., the food on which fungus gardens grow) and environmental origin may influence the volatiles the fungus releases. We found compounds related to the environment, including naphthalene. We show that the volatile profiles of fungal strains grown by Atta cephalotes are most similar to each other, whereas the profile of the fungus grown by ants in the genus Cyphomyrmex is more similar to that of their substrate than to the profiles of other cultivated fungi. Regarding cuticular hydrocarbons, we found that ants collected in the same location have more similar hydrocarbon profiles than ants of the same species collected in a different location, even if all the colonies had been maintained under the same conditions (temperature, substrate) for extended periods. Our results provide strong evidence that a combination of species genetics and environmental factors shape variations in the volatile chemical profiles of cultivated fungi. After long homogenization, ants still demonstrate a solid difference among the cuticular profiles.

Downloads

Download data is not yet available.

References

Adams, R. (2017) Identification of Essential Oil Components by Gas Chromatography / Mass Spectrometry, 4.1

Adams, R.M.M., Liberti, J., Illum, A.A., et al (2013) Chemically armed mercenary ants protect fungus-farming societies. PNAS 110: 15752–15757. DOI: https://doi.org/10.1073/pnas.1311654110

Ali, M., & Morgan, E. (1990). Chemical communication in insect communities: a guide to insect pheromones with special emphasis on social insects. Biological Reviews, 65: 227-247. DOI: https://doi.org/10.1111/j.1469-185X.1990.tb01425.x

Anderson, M.J. (2017). Permutational Multivariate Analysis of Variance (PERMANOVA). Wiley StatsRef: Statistics Reference Online.

Anderson, M.J. (2006). Distance-based tests for homogeneity of multivariate dispersions. Biometrics 62: 245-253. DOI: https://doi.org/10.1111/j.1541-0420.2005.00440.x

Anderson, M.J. (2001). A new method for non-parametric multivariate analysis of variance. Austral Ecology 26: 32-46. DOI: https://doi.org/10.1111/j.1442-9993.2001.01070.pp.x

Billen, J. & Morgan, E.D. (1998). Pheromone communication in social insects: sources and secretions. 1st ed. CRC Press. 31p.

Blomquist, G. (2010). Structure and analysis of insect hydrocarbons. In: Blomquist G, Bagnères A (eds) Insect hydrocarbons: biology, biochemistry and Physiology. Cambridge University Press, pp 19-34. DOI: https://doi.org/10.1017/CBO9780511711909.003

Branstetter, M.G., Longino, J.T., Ward, P.S., Faircloth, B.C. (2017) Enriching the ant tree of life: enhanced UCE bait set for genome-scale phylogenetics of ants and other Hymenoptera. Methods in Ecology and Evolution 8: 768-776. DOI: https://doi.org/10.1111/2041-210X.12742

Bray, J.R. & Curtis, J.T. (1957) An Ordination of the Upland Forest Communities of Southern Wisconsin. Ecological Monographs, 27: 325-349. DOI: https://doi.org/10.2307/1942268

Bretz, F., Hothorn, T., & Westfall, P. (2016) Multiple comparisons using R. CRC Press. DOI: https://doi.org/10.1201/9781420010909

Buckpitt, A., Kephalopoulos, S., Koistinen K, et al. (2010) Naphthalene. World Health Organization.

Buttigieg, P.L. & Ramette, A. (2014) A guide to statistical analysis in microbial ecology: a community-focused, living review of multivariate data analyses. FEMS Microbiology and Ecology, 90: 543-550. DOI: https://doi.org/10.1111/1574-6941.12437

Chapela, I.H., Rehner, S.A., Schultz, T.R., &Mueller, U.G. (1994) Evolutionary history of the symbiosis between fungus-growing ants and their fungi. Science, 266: 1691-1694. DOI: https://doi.org/10.1126/science.266.5191.1691

Clarke, K.R. (1993) Non-parametric multivariate analyses of changes in community structure. Australian Journal of Ecology, 18: 117-143. DOI: https://doi.org/10.1111/j.1442-9993.1993.tb00438.x

Copren, K.A., Nelson, L.J., Vargo, E.L., & Haverty, M.I. (2005) Phylogenetic analyses of mtDNA sequences corroborate taxonomic designations based on cuticular hydrocarbons in subterranean termites. Molecular Phylogenetics and Evolution 35:689-700. DOI: https://doi.org/10.1016/j.ympev.2005.03.002

Cremer, S., Pull, C.D., & Fürst, M.A. (2018) Social immunity: emergence and evolution of colony-level disease protection. Annual Review of Entomology, 63:105-123. DOI: https://doi.org/10.1146/annurev-ento-020117-043110

Currie, C.R. (2001) A community of ants, fungi and bacteria: A multilateral approach to studying symbiosis. Annual Review of Microbiology, 55: 357–380. DOI: https://doi.org/10.1146/annurev.micro.55.1.357

Cuvillier-Hot, V., Cobb, M., Malosse, C., & Peeters, C. (2001) Sex, age and ovarian activity affect cuticular hydrocarbons in Diacamma ceylonense, a queenless ant. Journal of Insect Physiology, 47: 485–493. DOI: https://doi.org/10.1016/S0022-1910(00)00137-2

Daisy, B.H., Strobel, G.A., Castillo, U., et al (2002) Naphthalene, an insect repellent, is produced by Muscodor vitigenus, a novel endophytic fungus. Microbiology 148: 3737–3741. DOI: https://doi.org/10.1099/00221287-148-11-3737

D’Ettorre, P., Heinze, J., Schulz, C., et al (2004) Does she smell like a queen? Chemoreception of a cuticular hydrocarbon signal in the ant Pachycondyla inversa. Journal of Experimental Biology, 207: 1085–1091. DOI: https://doi.org/10.1242/jeb.00865

Dickschat, J.S. (2017) Fungal volatiles – a survey from edible mushrooms to moulds. Natural Product Reports, 34: 310–328. DOI: https://doi.org/10.1039/C7NP00003K

Ditzen, M., Pellegrino, M., & Vosshall, L.B. (2008) Insect odorant receptors are molecular targets of the insect repellent DEET. Science, 319: 1838–1842. DOI: https://doi.org/10.1126/science.1153121

Dronnet, S., Lohou, C., Christides, J.P., & Bagnères, A.G. (2006). Cuticular hydrocarbon composition reflects genetic relationship among colonies of the introduced termite Reticulitermes santonensis feytaud. Journal of Chemical Ecology, 32: 1027-1042. DOI: https://doi.org/10.1007/s10886-006-9043-x

Ferguson-Gow, H., Sumner, S., Bourke. A.F.G., et al (2014) Colony size predicts division of labour in attine ants: Colony size predicts division of labour in attine ants. Proceedings of the Royal Society B: Biological Sciences 281: 1-9. DOI: https://doi.org/10.1098/rspb.2014.1411

Gond, S.K., Mishra, A., Sharma, V.K., et al. (2013). Isolation and characterization of antibacterial naphthalene derivative from Phoma herbarum, an endophytic fungus of Aegle marmelos. Current Science, 105: 167-169.

Gotelli, N.J. & Ellison, A.M. (2004). A primer of ecological statistics. ArtMed.

Hand, D.J., & Taylor, C.C. (1987). Multivariate analysis of variance and repeated measures: a practical approach for behavioural scientists. CRC Press. DOI: https://doi.org/10.1007/978-94-009-3143-5

Haverty, M.I., Page, M., Nelson, L.J., & Blomquist, G.J. (1988) Cuticular hydrocarbons of dampwood termites, Zootermopsis: Intra- and intercolony variation and potential as taxonomic characters. Journal of Chemical Ecology, 14: 1035-1058. DOI: https://doi.org/10.1007/BF01018791

Herz, H., Hölldobler, B., & Roces, F. (2008) Delayed rejection in a leaf-cutting ant after foraging on plants unsuitable for the symbiotic fungus. Behavioral Ecology, 19: 575-582. DOI: https://doi.org/10.1093/beheco/arn016

Hölldobler, B.W. & Wilson, E. (1990) The Ants. Belknap Press of Harvard University Press. DOI: https://doi.org/10.1007/978-3-662-10306-7

Howard, R.W. & Blomquist, G.J. (1982) Chemical ecology and biochemistry of insect hydrocarbons. Annual Review of Entomology, 27: 149–172. DOI: https://doi.org/10.1146/annurev.en.27.010182.001053

Howard, R.W. & Blomquist, G.J. (2005) Ecological, behavioral, and biochemical aspects of insect hydrocarbons. Annual Review of Entomology, 50: 371–393. DOI: https://doi.org/10.1146/annurev.ento.50.071803.130359

Kather. R. & Martin, S.J. (2015) Evolution of cuticular hydrocarbons in the hymenoptera: a meta-analysis. Journal of Chemical Ecology, 41: 871–883. DOI: https://doi.org/10.1007/s10886-015-0631-5

Le Conte, Y. & Hefetz, A. (2008) Primer pheromones in social hymenoptera. Annual Review of Entomology, 53: 523–542. DOI: https://doi.org/10.1146/annurev.ento.52.110405.091434

Legendre, P. & Legendre, L. (2012). Numerical ecology. Elsevier.

Leonhardt, S.D., Menzel, F., Nehring, V., & Schmitt, T. (2016) Ecology and evolution of communication in social insects. Cell, 164: 1277–1287. DOI: https://doi.org/10.1016/j.cell.2016.01.035

Leonhardt, S.D., Rasmussen, C., & Schmitt, T. (2013) Genes versus environment: geography and phylogenetic relationships shape the chemical profiles of stingless bees on a global scale. Proceedings of the Royal Society B, 280: 20130680. DOI: https://doi.org/10.1098/rspb.2013.0680

Li, H., Sosa-Calvo, J., Horn. H.A., Pupo, M.T., Clardy, J., Rabeling, C., Schultz, T.R., & Currie, C.R. (2018) Convergent evolution of complex structures for ant-bacterial defensive symbiosis in fungus-farming ants. Proceedings of the National Academy of Sciences, 115: 10720-10725. DOI: https://doi.org/10.1073/pnas.1809332115

Liang, D., Silverman, J. (2000) “You are what you eat”: diet modifies cuticular hydrocarbons and nestmate recognition in the Argentine ant, Linepithema humile. Die Naturwissenschaften, 87: 412–416. DOI: https://doi.org/10.1007/s001140050752

Little, A.E., Murakami, T., Mueller, U.G., & Currie, C.R. (2006) Defending against parasites: fungus-growing ants combine specialized behaviours and microbial symbionts to protect their fungus gardens. Biology Letters, 2: 12–16. DOI: https://doi.org/10.1098/rsbl.2005.0371

Liu, Z.B., Bagnères, A.G., Yamane, S., et al (2001) Intra-colony, inter-colony and seasonal variations of cuticular hydrocarbon profiles in Formica japonica (Hymenoptera, Formicidae). Insectes Sociaux, 48: 342–346. DOI: https://doi.org/10.1007/PL00001787

Martin, S. & Drijfhout, F. (2009a) A review of ant cuticular hydrocarbons. Journal of Chemical Ecology. 35: 1151–1161. DOI: https://doi.org/10.1007/s10886-009-9695-4

Martin, S.J. & Drijfhout, F.P. (2009b) How reliable is the analysis of complex cuticular hydrocarbon profiles by multivariate statistical methods? Journal of Chemical Ecology, 35: 375–382. DOI: https://doi.org/10.1007/s10886-009-9610-z

Mehdiabadi, N.J., & Schultz, T.R. (2009) Natural history and phylogeny of the fungus-farming ants (Hymenoptera: Formicidae). Myrmecological News, 13: 37–55.

Méndez-Bravo, A., Cortazar-Murillo, E.M., Guevara-Avendaño, E,. et al (2018) Plant growth-promoting rhizobacteria associated with avocado display antagonistic activity against Phytophthora cinnamomi through volatile emissions. PLOS ONE, 13:e0194665. DOI: https://doi.org/10.1371/journal.pone.0194665

Menzel, F., Blaimer, B.B., & Schmitt, T. (2017a) How do cuticular hydrocarbons evolve? Physiological constraints and climatic and biotic selection pressures act on a complex functional trait. Proceedings of the Royal Society B, 284: 20161727. DOI: https://doi.org/10.1098/rspb.2016.1727

Menzel, F., Schmitt, T., & Blaimer, B.B. (2017b) The evolution of a complex trait: cuticular hydrocarbons in ants evolveindependent from phylogenetic constraints. Journal of Evolutionary Biology, 30: 1372–1385. DOI: https://doi.org/10.1111/jeb.13115

Morath, S.U., Hung, R., & Bennett, J.W. (2012) Fungal volatile organic compounds: A review with emphasis on their biotechnological potential. Fungal Biology Reviews, 26: 73–83. DOI: https://doi.org/10.1016/j.fbr.2012.07.001

Moreira, A., Forti, L., Boaretto, M., et al. (2004) External and internal structure of Atta bisphaerica Forel (Hymenoptera: Formicidae) nests. Journal of Applied Entomology, 128: 204–211. DOI: https://doi.org/10.1111/j.1439-0418.2004.00839.x

Mueller, U.G., Schultz, T.R., Currie, C.R., et al (2001) The origin of the attine ant-fungus mutualism. The Quarterly Review of Biology, 76: 169-97 DOI: https://doi.org/10.1086/393867

National Institute of Standards and Technology (2017) NIST Chemical web book. In: Nist web book. https://webbook.nist. gov/chemistry/. Accessed 25 Sep 2018.

Németh, É., Bernáth, J., & Héthelyi, É. (1993) Diversity in chemotype reaction affected by ontogenetical and ecological factors. In: International Symposium on Medicinal and Aromatic Plants 344. pp 178–187. DOI: https://doi.org/10.17660/ActaHortic.1993.344.21

North, R.D., Jackson, C.W., & Howse, P.E. (1999) Communication between the fungus garden and workers of the leaf-cutting ant, Atta sexdens rubropilosa, regarding choice of substrate for the fungus. Physiological Entomology, 24: 127133. DOI: https://doi.org/10.1046/j.1365-3032.1999.00122.x

Oksanen, J., Blanchet, F.G., Kindt, R., et al (2017). Vegan: Community Ecology Package. R Package Version 2.2-0. http://CRAN.Rproject.org/package=vegan

Parry, K. & Morgan, E.D. (1979) Pheromones of ants: a review. Physiological Entomology, 4: 161–189. DOI: https://doi.org/10.1111/j.1365-3032.1979.tb00193.x

Pasanen, P., Korpi, A., Kalliokoski, P., & Pasanen, A-L. (1997) Growth and volatile metabolite production of Aspergillus versicolor in house dust. Environment International, 23: 425–432. DOI: https://doi.org/10.1016/S0160-4120(97)00027-5

R Foundation for Statistical Computing (2016) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Viena, Austria.

Richard, F.J., Hefetz, A., Christides, J.P., & Errard, C. (2004) Food influence on colonial recognition and chemical signature between nestmates in the fungus-growing ant Acromyrmex subterraneus subterraneus. Chemoecology, 14: 9–16. DOI: https://doi.org/10.1007/s00049-003-0251-3

Richard, F.J., Poulsen, M., Hefetz, A, et al (2007) The origin of the chemical profiles of fungal symbionts and their significance for nestmate recognition in Acromyrmex leaf-cutting ants. Behavioral Ecology and Sociobiology, 61: 1637–1649. DOI: https://doi.org/10.1007/s00265-007-0395-1

Ridley, P., Howse, P.E., & Jackson, C.W. (1996) Control of the behaviour of leaf-cutting ants by their “symbiotic” fungus. Experientia, 52: 631–635. DOI: https://doi.org/10.1007/BF01969745

Schultz, T.R. & Brady, S.G. (2008) Major evolutionary transitions in ant agriculture. PNAS, 105: 5435–5440. DOI: https://doi.org/10.1073/pnas.0711024105

Shik, J.Z., Gomez, E.B., Kooij, P.W., et al (2016) Nutrition mediates the expression of cultivar–farmer conflict in a fungus-growing ant. PNAS, 113: 201606128. DOI: https://doi.org/10.1073/pnas.1606128113

Soroker, V., Lucas, C., Simon, T., et al (2003) Hydrocarbon distribution and colony odour homogenisation in Pachycondyla apicalis. Insectes Sociaux, 50: 212–217. DOI: https://doi.org/10.1007/s00040-003-0669-1

Tavares, M., da Silva, M.R.M., de Siqueira, L.B. de O, et al (2018) Trends in insect repellent formulations: A review. International Journal of Pharmaceutics, 539: 190–209. DOI: https://doi.org/10.1016/j.ijpharm.2018.01.046

Valadares, L, Nascimento, D., & Nascimento, F.S. (2015) Foliar substrate affects cuticular hydrocarbon profiles and intraspecific aggression in the leafcutter ant Atta sexdens. Insects, 6: 141–151. DOI: https://doi.org/10.3390/insects6010141

Valadares, L. & Nascimento, F.S. (2016) Chemical cuticular signature of leafcutter ant Atta sexdens (Hymenoptera, Formicidae) worker subcastes. Revista Brasileira de Entomologia, 60: 308-311. DOI: https://doi.org/10.1016/j.rbe.2016.06.008

van Wilgenburg, E., Symonds, M.R.E., & Elgar, M.A. (2011) Evolution of cuticular hydrocarbon diversity in ants. Journal of Evolutionary Biology, 24: 1188-1198. DOI: https://doi.org/10.1111/j.1420-9101.2011.02248.x

Viana, A.M.M., Frézard, A., & Malosse, C, et al (2001) Colonial recognition of fungus in the fungus-growing ant Acromyrmex subterraneus subterraneus (Hymenoptera: Formicidae). Chemoecology, 11: 29-36. DOI: https://doi.org/10.1007/PL00001829

Weber, N. (1958) Evolution in fungus-growing ants. Proceedings of the Tenth International Congress of ldots, 2: 459-473.

Wilson, E.O. (1980) Caste and division of labor in leaf-cutting ants (Hymenoptera: Formicidae: Atta). Behavioral Ecology and Sociobiology, 7: 143-165. DOI: https://doi.org/10.1007/BF00299521

Downloads

Published

2025-01-30

How to Cite

Assis, D. S., Schultz, T., Brodowski, S., Newsome, G. A., & Nascimento, F. S. do. (2025). The Laboratory Environment Affects the Volatiles of Fungus Gardens in the Colonies of Fungus-farming Ants. Sociobiology, 72(1), e7968. https://doi.org/10.13102/sociobiology.v72i1.9768

Issue

Section

Research Article - Ants