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      Single-Cell DNA Barcoding Using Sequences from the Small Subunit rRNA and Internal Transcribed Spacer Region Identifies New Species of Trichonympha and Trichomitopsis from the Hindgut of the Termite Zootermopsis angusticollis

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          Abstract

          To aid in their digestion of wood, lower termites are known to harbour a diverse community of prokaryotes as well as parabasalid and oxymonad protist symbionts. One of the best-studied lower termite gut communities is that of Zootermopsis angusticollis which has been known for almost 100 years to possess 3 species of Trichonympha ( T. campanula, T. collaris, and T. sphaerica), 1 species of Trichomitopsis ( T. termopsidis), as well as smaller flagellates. We have re-assessed this community by sequencing the small subunit (SSU) rRNA gene and the internal transcribed spacer (ITS) region from a large number of single Trichonympha and Trichomitopsis cells for which morphology was also documented. Based on phylogenetic clustering and sequence divergence, we identify 3 new species: Trichonympha postcylindrica, Trichomitopsis minor, and Trichomitopsis parvus spp. nov. Once identified by sequencing, the morphology of the isolated cells for all 3 new species was re-examined and found to be distinct from the previously described species: Trichonympha postcylindrica can be morphologically distinguished from the other Trichonympha species by an extension on its posterior end, whereas Trichomitopsis minor and T. parvus are smaller than T. termopsidis but similar in size to each other and cannot be distinguished based on morphology using light microscopy. Given that Z. angusticollis has one of the best characterized hindgut communities, the near doubling of the number of the largest and most easily identifiable symbiont species suggests that the diversity of hindgut symbionts is substantially underestimated in other termites as well. Accurate descriptions of the diversity of these microbial communities are essential for understanding hindgut ecology and disentangling the interactions among the symbionts, and molecular barcoding should be a priority for these systems.

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          Author and article information

          Contributors
          Role: Editor
          Journal
          PLoS One
          PLoS ONE
          plos
          plosone
          PLoS ONE
          Public Library of Science (San Francisco, USA )
          1932-6203
          2013
          11 March 2013
          : 8
          : 3
          : e58728
          Affiliations
          [1 ]Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada
          [2 ]Department of Biology, University of Victoria, Victoria, British Columbia, Canada
          Université Paris Sud, France
          Author notes

          Competing Interests: The authors have declared that no competing interests exist.

          Conceived and designed the experiments: VT PJK. Performed the experiments: VT ERJ. Analyzed the data: VT. Contributed reagents/materials/analysis tools: PJK. Wrote the paper: VT SJP PJK.

          Article
          PONE-D-12-36304
          10.1371/journal.pone.0058728
          3594152
          23536818
          a010c796-487c-47e3-a7e6-1310fed9ad73
          Copyright @ 2013

          This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

          History
          : 20 November 2012
          : 5 February 2013
          Page count
          Pages: 12
          Funding
          This work was supported by a grant (227301) from the Natural Sciences and Engineering Resarch Council of Canada ( www.nserc.ca). VT was funded through the Junior Fellow Academy of the Canadian Institute for Advanced Research (CIFAR) ( www.cifar.ca). PJK and SJP are CIFAR fellows. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
          Categories
          Research Article
          Biology
          Ecology
          Biodiversity
          Evolutionary Biology
          Evolutionary Processes
          Coevolution
          Speciation
          Organismal Evolution
          Eukaryotic Evolution
          Microbial Evolution
          Microbiology
          Protozoology
          Protozoan Classification
          Microbial Ecology
          Microbial Evolution
          Zoology
          Entomology

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