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Characterization of Actinobacteria Associated with Three Ant–Plant Mutualisms

  • Invertebrate Microbiology
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Abstract

Ant–plant mutualisms are conspicuous and ecologically important components of tropical ecosystems that remain largely unexplored in terms of insect-associated microbial communities. Recent work has revealed that ants in some ant–plant systems cultivate fungi (Chaetothyriales) within their domatia, which are fed to larvae. Using Pseudomyrmex penetrator/Tachigali sp. from French Guiana and Petalomyrmex phylax/Leonardoxa africana and Crematogaster margaritae/Keetia hispida, both from Cameroon, as models, we tested the hypothesis that ant–plant–fungus mutualisms co-occur with culturable Actinobacteria. Using selective media, we isolated 861 putative Actinobacteria from the three systems. All C. margaritae/K. hispida samples had culturable Actinobacteria with a mean of 10.0 colony forming units (CFUs) per sample, while 26 % of P. penetrator/Tachigali samples (mean CFUs 1.3) and 67 % of P. phylax/L. africana samples (mean CFUs 3.6) yielded Actinobacteria. The largest number of CFUs was obtained from P. penetrator workers, P. phylax alates, and C. margaritae pupae. 16S rRNA gene sequencing and phylogenetic analysis revealed the presence of four main clades of Streptomyces and one clade of Nocardioides within these three ant–plant mutualisms. Streptomyces with antifungal properties were isolated from all three systems, suggesting that they could serve as protective symbionts, as found in other insects. In addition, a number of isolates from a clade of Streptomyces associated with P. phylax/L. africana and C. margaritae/K. hispida were capable of degrading cellulose, suggesting that Streptomyces in these systems may serve a nutritional role. Repeated isolation of particular clades of Actinobacteria from two geographically distant locations supports these isolates as residents in ant–plant–fungi niches.

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Acknowledgments

We thank Erik Houck, Karen Bednar, Diana Xiao, and Dan Phillips for their assistance with lab work, and Gina Lewin and Adam Book for assistance in setting up the cellulose degradation assays. We thank Heidi Horn, Jonathan Klassen, and Charles Mason for technical suggestions and comments on this manuscript. ASH was partially funded by National Institutes of Health T32 GM07215-37. CRC received funding from National Science Foundation MCB-0702025 and DOE BER Office of Science DE-FC02-07ER64494.

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Authors and Affiliations

  1. Department of Bacteriology, University of Wisconsin, Madison, WI, 53706, USA

    Alissa S. Hanshew, Bradon R. McDonald & Cameron R. Currie

  2. School of Science and Technology, Universidad del Este, Carolina, Puerto Rico

    Carol Díaz Díaz

  3. REU-Microbiology, Department of Bacteriology, University of Wisconsin, Madison, WI, 53706, USA

    Carol Díaz Díaz

  4. Laboratory of Zoology, Faculty of Science, University of Yaoundé I, PO Box 812, Yaoundé, Cameroon

    Champlain Djiéto-Lordon

  5. CEFE, CNRS UMR 5175, 1919 route de Mende, 34293, Montpellier cedex 5, France

    Rumsaïs Blatrix

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  1. Alissa S. Hanshew
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  3. Carol Díaz Díaz
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  4. Champlain Djiéto-Lordon
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  6. Cameron R. Currie
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Correspondence to Cameron R. Currie.

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Supplemental Fig. 1

Representatives of bioassay evaluation of antifungal activity. Nineteen isolates were tested for their antifungal activity. a Trichoderma reesei control plate after complete growth over the full plate. b Nocardiodies sp. LaPpAH12 with no ZOI against T. reesei. c Streptomyces sp PsTaAH5 with a moderate ZOI against T. reesei. d Streptomyces sp KhCrAH316 with moderate ZOI against T. reesei. e Streptomyces sp PsTaAH124 with almost complete suppression of T. reesei (GIF 102 kb)

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Hanshew, A.S., McDonald, B.R., Díaz Díaz, C. et al. Characterization of Actinobacteria Associated with Three Ant–Plant Mutualisms. Microb Ecol 69, 192–203 (2015). https://doi.org/10.1007/s00248-014-0469-3

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