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      Metabarcoding reveals a high diversity of woody host-associated Phytophthora spp. in soils at public gardens and amenity woodlands in Britain

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          Abstract

          Forests and woodlands worldwide are being severely impacted by invasive Phytophthora species, with initial outbreaks in some cases occurring on host trees located in public parks and gardens. These highly disturbed sites with diverse planting practices may indeed act as harbours for invasive Phytophthora pathogens which are particularly well adapted to surviving in soil. High throughput Illumina sequencing was used to analyse Phytophthora species diversity in soil samples collected from 14 public garden/amenity woodland sites in northern Britain. Bioinformatic analyses revealed some limitations to using internal transcribed spacer as the barcode region; namely reporting of false positives and ambiguous species matches. Taking this into account, 35 distinct sequences were amplified across the sites, corresponding to 23 known Phytophthora species as well as twelve oomycete sequences with no match to any known Phytophthora species. Phytophthora pseudosyringae and P. austrocedri, both of which cause serious damage to trees and are regarded as fairly recent introductions to Britain, were the two most abundant Phytophthora species detected. There was no evidence that any of the detected Phytophthora species were more associated with any one type of host, healthy or otherwise. This study has demonstrated the ubiquity and diversity of Phytophthora species endemic in highly managed, extensively planted soil environments in Britain. Suggested improvements to the methodology and the practical implications of the findings in terms of mitigating Phytophthora spread and impact are discussed.

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          AMPLIFICATION AND DIRECT SEQUENCING OF FUNGAL RIBOSOMAL RNA GENES FOR PHYLOGENETICS

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            Swarm: robust and fast clustering method for amplicon-based studies

            Popular de novo amplicon clustering methods suffer from two fundamental flaws: arbitrary global clustering thresholds, and input-order dependency induced by centroid selection. Swarm was developed to address these issues by first clustering nearly identical amplicons iteratively using a local threshold, and then by using clusters’ internal structure and amplicon abundances to refine its results. This fast, scalable, and input-order independent approach reduces the influence of clustering parameters and produces robust operational taxonomic units.
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              PacBio metabarcoding of Fungi and other eukaryotes: errors, biases and perspectives.

              Second-generation, high-throughput sequencing methods have greatly improved our understanding of the ecology of soil microorganisms, yet the short barcodes (  500-bp barcode for reliable identification or when phylogenetic approaches are intended.
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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                PeerJ
                PeerJ
                PeerJ
                PeerJ Inc. (San Diego, USA )
                2167-8359
                16 May 2019
                2019
                : 7
                : e6931
                Affiliations
                [1 ]Forest Research , Roslin, Midlothian, UK
                [2 ]The Woodland Trust , Edinburgh, UK
                [3 ]Forest Research , Farnham, Surrey, UK
                [4 ]James Hutton Institute , Dundee, UK
                [5 ]School of Medicine, University of St. Andrews , St Andrews, UK
                [6 ]Institute of Evolutionary Biology, University of Edinburgh , Edinburgh, UK
                Author information
                http://orcid.org/0000-0002-9154-7954
                http://orcid.org/0000-0002-8392-2822
                http://orcid.org/0000-0001-9771-543X
                Article
                6931
                10.7717/peerj.6931
                6526010
                31143546
                796f07fa-9469-4dd3-b1e8-2a722fb03c5b
                © 2019 Riddell et al.

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                History
                : 13 November 2018
                : 9 April 2019
                Funding
                Funded by: Forestry Commission Scotland
                Award ID: SLA-14/15-034
                Funded by: Living With Environmental Change Phase 3 project ‘Phyto-Threats’ as part of the Tree Health and Plant Biosecurity Initiative
                Award ID: BB/N023463/1
                Funded by: European Union’s Horizon 2020 research and innovation programme POnTE (Pest Organisms Threatening Europe)
                Award ID: 635646
                Funded by: David E.L. Cooke, Pete E. Hedley, Leighton Pritchard, Peter Thorpe also received funding from the Scottish Government
                This work was supported by Forestry Commission Scotland (grant number SLA-14/15-034), the Living With Environmental Change Phase 3 project ‘Phyto-Threats’ as part of the Tree Health and Plant Biosecurity Initiative (grant number BB/N023463/1) and the European Union’s Horizon 2020 research and innovation programme POnTE (Pest Organisms Threatening Europe) (grant number 635646). David E.L. Cooke, Pete E. Hedley, Leighton Pritchard, Peter Thorpe also received funding from the Scottish Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Molecular Biology
                Mycology
                Plant Science

                phytophthora,metabarcoding,its1 barcode,illumina sequencing,soil,species diversity

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