Abstract

This study reveals the diversity and distribution of two major ubiquitous groups of soil amoebae, the genus Acanthamoeba and the Myxomycetes (plasmodial slime-moulds) that are rarely, if ever, recovered in environmental sampling studies. We analyzed 150 grassland soil samples from three Biodiversity Exploratories study regions in Germany. We developed specific primers targeting the V2 variable region in the first part of the small subunit of the ribosomal RNA gene for high-throughput pyrotag sequencing. From ca. 1 million reads, applying very stringent filtering and clustering parameters to avoid overestimation of the diversity, we obtained 273 acanthamoebal and 338 myxomycete operational taxonomic units (OTUs, 96% similarity threshold). This number is consistent with the genetic diversity known in the two investigated lineages, but unequalled to date by any environmental sampling study. Only very few OTUs were identical to already known sequences. Strikingly different OTUs assemblages were found between the three German regions (PerMANOVA p.value = 0.001) and even between sites of the same region (multiple-site Simpson-based similarity indices <0.4), showing steep biogeographical gradients.

Highlights

  • This study reveals the diversity and distribution of two major ubiquitous groups of soil amoebae, the genus Acanthamoeba and the Myxomycetes that are rarely, if ever, recovered in environmental sampling studies

  • Using BLAST, 87% of these could be assigned to Acanthamoeba or Myxomycetes reference sequences

  • Protophysarum phloiogenum, a Physarida, was mis-assigned to Stemonitida, probably due to a long-branch attraction artefact. For both Acanthamoeba and Myxomycetes, we provide a complete list of the Operational taxonomic units (OTUs), their taxonomic affiliation according to BLAST and their occurrences in each soil sample (Table S4)

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Summary

Introduction

This study reveals the diversity and distribution of two major ubiquitous groups of soil amoebae, the genus Acanthamoeba and the Myxomycetes (plasmodial slime-moulds) that are rarely, if ever, recovered in environmental sampling studies. 1 million reads, applying very stringent filtering and clustering parameters to avoid overestimation of the diversity, we obtained 273 acanthamoebal and 338 myxomycete operational taxonomic units (OTUs, 96% similarity threshold) This number is consistent with the genetic diversity known in the two investigated lineages, but unequalled to date by any environmental sampling study. World wide inventories have been established based on the systematic collection of such fruiting bodies (listed in http://eumycetozoa.com/data/genera.php, updated June 11, 2014)[25] Because they have been classified as fungi for a long time, Myxomycetes are excluded from amoebozoan treatises[26] and from most soil inventories[5]. Providing that specific molecular probes are used, dark-spore Myxomycetes can be recovered from www.nature.com/scientificreports/

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