Fungal Communities In The Garden Chamber Soils Of Leaf-cutting Ants

Leaf-cutting ants modify the properties of the soil adjacent to their nests. Here, we examined whether such an ant-altered environment impacts the belowground fungal communities. Fungal diversity and community structure of soil from the fungus garden chambers of Atta sexdens rubropilosa and Atta bisphaerica, two widespread leaf-cutting ants in Brazil, were determined and compared with non-nest soils. Culture-dependent methods revealed similar species richness but different community compositions between both types of soils. Penicillium janthinellum and Trichoderma spirale were the prevalent isolates in fungus chamber soils and non-nest soils, respectively. In contrast to cultivation methods, analyses of clone libraries based on the internal transcribed spacer (ITS) region indicated that richness of operational taxonomic units significantly differed between soils of the fungus chamber and non-nest soils. FastUnifrac analyses based on ITS sequences further revealed a clear distinction in the community structure between both types of soils. Plectania milleri and an uncultured Clavariaceae fungus were prevalent in fungus chamber soils and non-nest soils, respectively. FastUnifrac analyses also revealed that fungal community structures of soil from the garden chambers markedly differed among ant species. Our findings suggest that leaf-cutting ants affect fungal communities in the soil from the fungus chamber in comparison to non-nest soils.541111861196Schultz, T.R., Brady, S.G., Major evolutionary transitions in ant agriculture (2008) Proc. Natl. Acad. Sci. USA, 105, pp. 5435-5440Weber, N.A., (1972) Gardening Ants: The Attines, , Memoirs of the American Philosophical Society, PhiladelphiaCaldera, E.J., Poulsen, M., Suen, G., Currie, C.R., Insect symbioses: a case study of past, present, and future fungus-growing ant research (2009) Environ. Entomol., 38, pp. 78-92Currie, C.R., Mueller, U.G., Malloch, D., The agricultural pathology of ant fungus gardens (1999) Proc. Natl. Acad. Sci. USA, 96, pp. 7998-8002Currie, C.R., Prevalence and impact of a virulent parasite on a tripartite mutualism (2001) Oecologia, 128, pp. 99-106Mueller, U.G., Symbiont recruitment versus ant-symbiont co-evolution in the attine ant-microbe symbiosys (2012) Curr. Opin. Microbiol., 15, pp. 269-277Little, A.E.F., Currie, C.R., Symbiotic complexity: discovery of a fifth symbiont in the attine ant-microbe symbiosis (2007) Biol. Lett., 3, pp. 501-504Little, A.E.F., Currie, C.R., Black yeasts symbionts compromise the efficiency of antibiotic defenses in fungus-growing ants (2008) Ecology, 89, pp. 1216-1222Bacci, M., Jr., Ribeiro, S.B., Casarotto, M.E.F., Pagnocca, F.C., Biopolymer-degrading bacteria from nests of the leaf-cutting ant Atta sexdens rubropilosa (1995) Braz. J. Med. Biol. Res., 28, pp. 79-82Scott, J.J., Budsberg, K.J., Suen, G., Wixon, D.L., Microbial community structure of leaf-cutter ant fungus-gardens and refuse dumps (2010) PLoS ONE, 5, p. e9992Rodrigues, A., Bacci, M., Jr., Mueller, U.G., Ortiz, A., Microfungal "weeds" in the leafcutter ant symbiosis (2008) Microb. Ecol., 56, pp. 604-614Rodrigues, A., Mueller, U.G., Ishak, H.D., Bacci, M., Jr., Ecology of microfungal communities in gardens of fungus-growing ants (Hymenoptera: Formicidae): a year-long survey of three species in Central Texas (2011) FEMS Microbiol. Ecol., 78, pp. 244-255Pinto-Tomás, A.A., Anderson, M.A., Suen, G., Stevenson, D.M., Symbiotic nitrogen fixation in the fungus gardens of leaf-cutter ants (2009) Science, 326, pp. 1120-1123Currie, C.R., Stuart, A.E., Weeding and grooming of pathogens in agriculture by ants (2001) Proc. R. Soc. B., 268, pp. 1033-1039Cremer, S., Armitage, S.A.O., Schmid-Hempel, P., Social immunity (2007) Curr. Biol., 17, pp. 693-702Vander Meer, R., Ant interactions with soil organisms and associated semiochemicals (2012) J. Chem. Ecol., 38, pp. 728-745Schoenian, I., Spiteller, M., Ghaste, M., Wirth, R., Chemical basis of the synergism and antagonism in microbial communities in the nests of leaf-cutting ants (2011) Proc. Natl. Acad. Sci. USA, 108, pp. 1955-1960Fernández-Marín, H., Zimmerman, J.K., Rehner, S.A., Wcislo, W.T., Active use of the metapleural glands by ants in controlling fungal infection (2006) Proc. R. Soc. B, 273, pp. 1689-1695Fernández-Marín, H., Zimmerman, J.K., Nash, D., Boomsma, J.J., Reduced biological control and enhanced chemical pest management in the evolution of fungus farming ants (2009) Proc. R. Soc. B, 276, pp. 2263-2269Farji-Brener, A.G., Leaf-cutting ant nests and soil biota abundance in a semi-arid steppe of northwestern Patagonia (2010) Sociobiology, 56, pp. 549-557Mora, P., Miambi, E., Jiménez, J.J., Decaëns, T., Functional complement of biogenic structures produced by earthworms, termites and ants in the neotropical savannas (2005) Soil Biol. Biochem., 37, pp. 1043-1048Holec, M., Frouz, J., The effect of two ant species Lasius niger and Lasius flavus on soil properties in two contrasting habitats (2006) Eur. J. Soil Biol., 42, pp. S213-S217Ba, A.S., Phillips, S.A., Anderson, J.T., Yeasts in mound soil of the red imported fire ant (2000) Mycol. Res., 104, pp. 969-973Baird, R., Woolfolk, S., Watson, C.E., Survey of bacterial associates of black/hybrid imported fire ants from mounds in Mississippi (2007) Southeast Natl., 6, pp. 615-632Zettler, J.A., McInnis, T.M., Allen, C.R., Spira, T.P., Biodiversity of fungi in red imported fire ant (Hymenoptera: Formicidae) mounds (2002) Ann. Entomol. Soc. Am., 95, pp. 487-491Folgarait, P., Ant biodiversity and its relationship to ecosystem functioning: a review (1998) Biodivers. Conserv., 7, pp. 1221-1244Hudson, T.M., Turner, B.L., Herz, H., Robinson, J.S., Temporal patterns of nutrient availability around nests of leaf-cutting ants (Atta colombica) in secondary moist tropical forest (2009) Soil Biol. Biochem., 41, pp. 1088-1093Jones, C.G., Lawton, J.H., Shachak, M., Organisms as ecosystem engineers (1994) Oikos, 69, pp. 373-386Jiménez, J.J., Decaëns, T., Chemical variations in the biostructures produced by soil ecosystem engineers. Examples from the neotropical savannas (2006) Eur. J. Soil Biol., 42, pp. S92-S102Jiménez, J.J., Decaëns, T., Lavelle, P., C and N concentrations in biogenic structures of a soil-feeding termite and a fungus-growing ant in the Colombian savannas (2008) Appl. Soil Ecol., 40, pp. 120-128Moutinho, P., Nepstad, D.C., Davidson, E.A., Influence of leaf-cutting ant nests on secondary forest growth and soil properties in Amazonia (2003) Ecology, 84, pp. 1265-1276Boulton, A.M., Jaffee, B.A., Scow, K.M., Effects of a common harvester ant (Messor andrei) on richness and abundance of soil biota (2003) Appl. Soil Ecol., 23, pp. 257-265Boulton, A.M., Amberman, K.D., How ants increase soil biota richness and abundance: a field experiment (2006) Biodivers. Conserv., 15, pp. 69-82Dauber, J., Niechoj, R., Baltruschat, H., Wolters, V., Soil engineering ants increase grass root arbuscular mycorrhizal colonization (2008) Biol. Fertil. Soils, 44, pp. 791-796Ginzburg, O., Whitford, W.G., Steinberger, Y., Effects of harvester ant (Messor spp.) activity on soil properties and microbial communities in a Negev Desert ecosystem (2008) Biol. Fertil. Soils, 45, pp. 165-173Ishak, H.D., Plowes, R., Sen, R., Kellner, K., Bacterial diversity in Solenopsis invicta and Solenopsis geminata ant colonies characterized by 16S amplicon 454 pyrosequencing (2011) Env. Microbiol., 61, pp. 821-831Dauber, J., Wolters, V., Microbial activity and functional diversity in the mounds of the three different ant species (2000) Soil. Biol. Biochem., 32, pp. 93-99Moreira, A.A., Forti, L.C., Boaretto, M.A.C., Andrade, A.P.P., External and internal structure of Atta bisphaerica Forel (Hymenoptera: Formicidae) nests (2004) J. Appl. Entomol., 128, pp. 204-211Rodrigues, A., Cable, R.N., Mueller, U.G., Bacci, M., Jr., Antagonistic interaction between garden yeasts and microfungal garden pathogens of leaf-cutting ants (2009) Anton. van Leeuwen., 96, pp. 331-342Domsch, K.H., Gams, W., Anderson, T., (1980) Compendium of Soil Fungi, , Academic Press, LondonSamson, R.A., Hoekstra, E.S., Frisvad, J.C., (2000) Introduction to Food-Airborne Fungi, , 6th edn., Centraalbureau voor Schimmelcultures, BaarnKlich, M.A., (2002) Identification of common Aspergillus species, , Centraalbureau voor Schimmelcultures, BaarnPitt, J.I., (2000) A Laboratory Guide to Common Penicillium Species, , 3rd edn., Commonwealth Scientific and Industrial Research Organization, North RydeSampaio, J.P., Gadanho, M., Santos, S., Duarte, F.L., Polyphasic taxonomy of the basidiomycetous yeast genus Rhodosporidium: Rhodosporidium kratochvilovae and related anamorphic species (2001) Int. J. Syst. Evol. Microbiol., 51, pp. 687-697Pagnocca, F.C., Rodrigues, A., Nagamoto, N.S., Bacci, M., Jr., Yeasts and filamentous fungi carried by the gynes of leaf-cutting ants (2008) Anton. van Leeuwen., 94, pp. 517-526Kurtzman, C.P., Robnett, C.J., Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequence (1998) Anton. van Leeuwen., 73, pp. 331-371White, T.J., Bruns, T., Lee, S., Taylor, J.W., Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics (1990) PCR Protocols: A Guide to Methods and Applications, pp. 315-322. , in: Innis, M.A., Gelfand, D.H., Sninsky, J.J., White, T.J., Eds.), Academic Press, New YorkHall, T.A., BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT (1999) Nucl. Acids Symp. Ser., 41, pp. 95-98Unterseher, M., Schnittler, M., Species richness analysis and ITS rDNA phylogeny revealed the majority of cultivable foliar endophytes from beech (Fagus sylvatica) (2010) Fungal Ecol., 3, pp. 366-378Durham, A.M., Kashiwabara, A.Y., Matsunaga, F.T., Ahagon, P.H., EGene: a configurable pipeline generation system for automated sequence analysis (2005) Bioinformatics, 21, pp. 2812-2813Nilsson, R.H., Abarenkov, K., Veldre, V., Nylinder, S., An open source chimera checker for the fungal ITS region (2010) Mol. Ecol. Res., 10, pp. 1076-1081Götz, S., García-Gómez, J.M., Terol, J., Williams, T.D., A high-throughput functional annotation and data mining with the Blast2GO suite (2008) Nucl. Acids Res., 36, pp. 3420-3435Colwell, R.K., (2009), http://purl.oclc.org/estimates, EstimateS: Statistical estimation of species richness and shared species from samples. Version 8.2. User's Guide and application published at:Edgar, R.C., Search and clustering orders of magnitude faster than BLAST (2010) Bioinformatics, 26, pp. 2460-2461Schloss, P.D., Westcott, S.L., Ryabin, T., Hall, J.R., Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities (2009) Appl. Env. Microbiol., 75, pp. 7537-7541Edgar, R.C., MUSCLE: multiple sequence alignment with high accuracy and high throughput (2004) Nucl. Acids Res., 32, pp. 1792-1797Tamura, K., Peterson, D., Peterson, N., Stecher, G., MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods (2011) Mol. Biol. Evol., 28, pp. 2731-2739Hamady, M., Lozupone, C., Knight, R., Fast UniFrac: facilitating high-throughput phylogenetic analyses of microbial communities including analysis of pyrosequencing and PhyloChip data (2010) ISME J., 4, pp. 17-27Price, M.N., Dehal, P.S., Arkin, A.P., FastTree: computing large minimum-evolution trees with profiles instead of a distance matrix (2009) Mol. Biol. Evol., 26, pp. 1641-1650Pagnocca, F.C., Masiulionis, V.E., Rodrigues, A., Specialized fungal parasites and opportunistic fungi in gardens of attine ants (2012) Psyche, 12, p. 9. , Article ID 905109Rodrigues, A., Pagnocca, F.C., Bacci, M., Jr., Hebling, M.J.A., Variability of non-mutualistic fungi associated with Atta sexdens rubropilosa nests (2005) Folia Microbiol., 50, pp. 421-425Bento, J.M.S., Della Lucia, T.M.C., Muchovej, R.M.C., Vilela, E.F., Influência da composição química e da população microbiana de diferentes horizontes do solo no estabelecimento de sauveiros iniciais de Atta laevigata (Hymenoptera: Formicidae) em laboratório (1991) An. Soc. Entomol. Bras., 20, pp. 307-317Rodrigues, O.D., (2007), Influência da comunidade microbiana do solo no estabelecimento de sauveiros iniciais de Atta sexdens rubropilosa Forel, 1908 (Hymenoptera: Formicidae). Dissertation, Escola Superior de Agricultura "Luiz de Queiroz", Universidade de São PauloSilva, A., Rodrigues, A., Bacci, M., Jr., Pagnocca, F.C., Susceptibility of the ant-cultivated fungus Leucoagaricus gongylophorus (Agaricales: Basidiomycota) towards microfungi (2006) Mycopathologia, 162, pp. 115-119Ortiz, A., Orduz, S., In vitro evaluation of Trichoderma and Gliocladium antagonism against the symbiotic fungus of the leaf-cutting ant Atta cephalotes (2001) Mycopathologia, 150, pp. 53-60Nagy, L.G., Petkovits, T., Kovács, G.M., Voigt, K., Where is the unseen fungal diversity hidden? A study of Mortierella reveals a large contribution of reference collections to the identification of fungal environmental sequences (2011) New Phytol., 191, pp. 789-794O'Brien, H.E., Parrent, J.L., Jackson, J.A., Moncalvo, J.M., Fungal community analysis by large-scale sequencing of environmental samples (2005) Appl. Env. Microbiol., 71, pp. 5544-555