Top 15 most abundant fungal classes represented in the sooty mould ITS-based pyrosequencing dataset.

<p>The fungal classes are in order of abundance decreasing from top to bottom, with unknown fungi taking the top two spots, followed by several classes from <i>Ascomycota</i> and <i>Basidiomycota</i>. Representative phylogenetic tree coloured by most abundant fungal classes and others coloured dark teal. Scale bar = 1.0 substitutions/site. Each sample is labelled with Sample IDs from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070316#pone-0070316-t001" target="_blank">Table 1</a>.</p

Host influence on the sooty mould community composition using different molecular and analysis methods.

<p>Samples of sooty mould from black beech and red beech were compared.</p>1<p>Test Statistic: Adonis = F-test statistic; MRPP = chance corrected within-group agreement, A.</p>2<p>Significance codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05.</p

Diverse Honeydew-Consuming Fungal Communities Associated with Scale Insects

<div><p>Sooty mould fungi are ubiquitous, abundant consumers of insect-honeydew that have been little-studied. They form a complex of unrelated fungi that coexist and compete for honeydew, which is a chemically complex resource. In this study, we used scanning electron microscopy in combination with T-RFLP community profiling and ITS-based tag-pyrosequencing to extensively describe the sooty mould community associated with the honeydews of two ecologically important New Zealand coelostomidiid scale insects, <i>Coelostomidia wairoensis</i> and <i>Ultracoelostoma brittini</i>. We tested the influence of host plant on the community composition of associated sooty moulds, and undertook limited analyses to examine the influence of scale insect species and geographic location. We report here a previously unknown degree of fungal diversity present in this complex, with pyrosequencing detecting on average 243 operational taxonomic units across the different sooty mould samples. In contrast, T-RFLP detected only a total of 24 different “species” (unique peaks). Nevertheless, both techniques identified similar patterns of diversity suggesting that either method is appropriate for community profiling. The composition of the microbial community associated with individual scale insect species varied although the differences may in part reflect variation in host preference and site. Scanning electron microscopy visualised an intertwined mass of fungal hyphae and fruiting bodies in near-intact physical condition, but was unable to distinguish between the different fungal communities on a morphological level, highlighting the need for molecular research. The substantial diversity revealed for the first time by pyrosequencing and our inability to identify two-thirds of the diversity to further than the fungal division highlights the significant gap in our knowledge of these fungal groups. This study provides a first extensive look at the community diversity of the fungal community closely associated with the keystone insect-honeydew systems of New Zealand’s native forests and suggests there is much to learn about sooty mould communities.</p></div

Species abundances per site

This data file contains abundances for all pollinator species at each sampling site used in response diversity analyses

Overall pattern of scale species influence on the composition of sooty mould community associated with <i>Ultracoelostoma brittini</i> and <i>Coelostomidia wairoensis</i> honeydews.

<p>Plot A shows the multidimensional scaling plot based on T-RFLP peak-profiles using Bray-Curtis distances with “+” indicating sooty mould samples from <i>U. brittini</i> (n = 19, samples pooled from all sites and hosts) and open triangles for those from <i>C. wairoensis</i> (n = 8); Plot B shows the principle coordinates analyses based on ITS-based pyrosequencing data using weighted Unifrac distances with grey circles indicating samples from <i>U. brittini</i> (n = 8, samples pooled from all sites and hosts) and those from <i>C. wairoensis</i> indicated by black squares <i>Coelostomidia wairoensis</i> (n = 4).</p

Sampling overview.

<p>Sampling overview.</p

Sampling site co-ordinates

This file contains coordinates for all pollinator sampling sites

Stavert et al. Code from Exotic species enhance response diversity to land-use change but modify functional composition

R script for functional redundancy and response diversity analyses used in the paper

Sooty moulds associated with New Zealand coelostomidiid honeydew.

<p>Scanning electron micrographs of sooty mould from A, kānuka infested with <i>Coelostomidia wairoensis</i> (scale bar = 300 µm); B, kānuka infested with <i>C. wairoensis</i>, showing several types of smooth surface and rough surface (with projections) hyphae of different sizes and shapes (scale bar = 20 µm); C, kānuka infested with <i>C. wairoensis</i> showing a large rough surface spore-containing structure (scale bar = 20 µm); D, kānuka infested with <i>C. wairoensis</i> showing fruiting bodies (scale bar = 20 µm) and E, black beech infested with <i>Ultracoelostoma brittini</i> showing a cluster of fruiting bodies (scale bar = 250 µm).</p

Scale insect species influence on the sooty mould community composition using different molecular and analysis methods.

<p>Samples of sooty mould from <i>Ultracoelostoma brittini</i> and <i>Coelostomidia wairoensis</i> were compared.</p>1<p>Test Statistic: Adonis = F-test statistic; MRPP = chance corrected within-group agreement, A.</p>2<p>Significance codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05.</p