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Culture-Free Survey Reveals Diverse and Distinctive Fungal Communities Associated with Developing Figs (Ficus spp.) in Panama

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Abstract

The ancient association of figs (Ficus spp.) and their pollinating wasps (fig wasps; Chalcidoidea, Hymenoptera) is one of the most interdependent plant–insect mutualisms known. In addition to pollinating wasps, a diverse community of organisms develops within the microcosm of the fig inflorescence and fruit. To better understand the multipartite context of the fig–fig wasp association, we used a culture-free approach to examine fungal communities associated with syconia of six species of Ficus and their pollinating wasps in lowland Panama. Diverse fungi were recovered from surface-sterilized flowers of all Ficus species, including gall- and seed flowers at four developmental stages. Fungal communities in syconia and on pollinating wasps were similar, dominated by diverse and previously unknown Saccharomycotina, and distinct from leaf- and stem endophyte communities in the same region. Before pollination, fungal communities were similar between gall- and seed flowers and among Ficus species. However, fungal communities differed significantly in flowers after pollination vs. before pollination, and between anciently diverged lineages of Ficus with active vs. passive pollination syndromes. Within groups of relatively closely related figs, there was little evidence for strict-sense host specificity between figs and particular fungal species. Instead, mixing of fungal communities among related figs, coupled with evidence for possible transfer by pollinating wasps, is consistent with recent suggestions of pollinator mixing within syconia. In turn, changes in fungal communities during fig development and ripening suggest an unexplored role of yeasts in the context of the fig–pollinator wasp mutualism.

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Acknowledgments

We gratefully acknowledge the National Science Foundation for supporting this research (IOB-062492 to AEA and an NSF Graduate Research Fellowship to EOM) as well as the Smithsonian Institute (Predoctoral Fellowship to EOM). We thank the Smithsonian Tropical Research Institute for logistical support and the government of Panama for permission to carry out this research. We are grateful to J. Hackett and A. Gomez for technical assistance, W. Marussich for collection of fig wasps, and J. U’Ren and V. Martinson for helpful discussion.

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Authors and Affiliations

  1. Department of Ecology and Evolutionary Biology, The University of Arizona, Tucson, AZ, 85721, USA

    Ellen O. Martinson

  2. Smithsonian Tropical Research Institute, Balboa, Ancon, Panama City, Republic of Panama

    Edward Allen Herre

  3. Department of Biology, The University of Maryland, College Park, MD, 20742, USA

    Carlos A. Machado

  4. School of Plant Sciences, The University of Arizona, Tucson, AZ, 85721, USA

    A. Elizabeth Arnold

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  1. Ellen O. Martinson
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  2. Edward Allen Herre
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  3. Carlos A. Machado
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Correspondence to A. Elizabeth Arnold.

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Supplemental Figure 1

Locations of trees sampled in Barro Colorado National Monument, Panama. 1 F. insipida, 2 F. maxima, 3 F. triangle, 4 F. costaricana, 5 F. popenoei, and 6 F. obtusifolia. Physically proximate trees harbored communities that were actually significantly less similar (comparison among trees 1–4; averaged JGR = 0.6891) than trees at further distances (trees 5–6 compared to trees 1–4; averaged JGR = 0.9804) (JPEG 1576 kb)

High resolution image (EPS 375 kb)

Supplemental Figure 2

Most likely tree resulting from RAxML analyses of the 5.8S and partial LSU, including results of maximum likelihood bootstrap (≥70 %; above branches) and Bayesian posterior probabilities (≥90 %, below branches), revealing the phylogenetic placement of fungal OTU recovered from figs in six species of Ficus in Panama. Tree was used for UniFrac analyses; clades are not collapsed, such that the position of each sequence can be examined. Leucosporidium antarcticum was included as an outgroup. Black bars indicate OTU based on 95 % sequence similarity (JPEG 5775 kb)

High resolution image (EPS 452 kb)

Supplemental Figure 3

OTU accumulation, 95 % confidence intervals, and bootstrap estimates of richness based on ITS-LSU OTU (95 % sequence similarity) for fungal communities sampled from each of six Ficus species (JPEG 2737 kb)

High resolution image (EPS 504 kb)

Supplemental Figure 4

OTU accumulation for fungi recovered from pollinating fig wasps, 95 % confidence intervals, and bootstrap estimates of richness based on the survey of six species of wasps in lowland Panama (JPEG 2618 kb)

High resolution image (EPS 432 kb)

Supplemental Figure 5

Most likely tree resulting from RAxML analyses of the 5.8S and partial LSU, including results of maximum likelihood bootstrap (≥70 %; above branches), revealing the phylogenetic placement of fungal OTU recovered from figs in six species of Ficus and their associated pollinating fig wasps in Panama. L. antarcticum was included as an outgroup. Black bars indicate OTU based on 95 % sequence similarity for Ficus sequences and red bars indicate sequences from their pollinating fig wasps (JPEG 4531 kb)

High resolution image (EPS 547 kb)

Supplemental Figure 6

Non-metric multidimensional scaling (NMDS) analysis of gall- and seed-flower fungal communities. Additional analysis shown in upper right corner was assessed by ANOSIM with the Morisita index based on non-singleton OTU (JPEG 241 kb)

High resolution image (EPS 376 kb)

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Martinson, E.O., Herre, E.A., Machado, C.A. et al. Culture-Free Survey Reveals Diverse and Distinctive Fungal Communities Associated with Developing Figs (Ficus spp.) in Panama. Microb Ecol 64, 1073–1084 (2012). https://doi.org/10.1007/s00248-012-0079-x

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