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Phylogenetic analysis of the distribution of deadly amatoxins among the little brown mushrooms of the genus Galerina Berbee, Mary; Landry, Brandon; Whitton, Jeannette; Bazzicalupo, Anna; Ceska, Oldriska

Description

Abstract

Some but not all of the species of 'little brown mushrooms' in the genus Galerina contain deadly amatoxins at concentrations equaling those in the death cap, Amanita phalloides. However, Galerina's ~300 species are notoriously difficult to identify by morphology and the identity of toxin-containing specimens has not been verified with DNA barcode sequencing. This left open the question of which Galerina species contain toxins and which do not. We selected specimens for toxin analysis using a preliminary phylogeny of the fungal DNA barcode region, the ribosomal internal transcribed spacer (ITS) region. Using liquid chromatography/mass spectrometry, we analyzed amatoxins from 70 samples of Galerina and close relatives, collected in western British Columbia, Canada. To put the presence of toxins into a phylogenetic context, we included the 70 samples in maximum likelihood analyses of 438 taxa, using ITS, RNA polymerase II second largest subunit gene (RPB2), and nuclear large subunit ribosomal RNA (LSU) gene sequences. We sequenced barcode DNA from types where possible to aid with applications of names. We detected amatoxins only in the 24 samples of the G. marginata s.l. complex in the Naucoriopsis clade. We delimited 56 putative Galerina species using Automatic Barcode Gap Detection software. Phylogenetic analysis showed moderate to strong support for Galerina infrageneric clades Naucoriopsis, Galerina, Tubariopsis, and Sideroides. Mycenopsis appeared paraphyletic and included Gymnopilus. Amatoxins were not detected in 46 samples from Galerina clades outside of Naucoriopsis or from outgroups. Our data show significant quantities of toxin in all mushrooms tested from the G. marginata s.l. complex. DNA barcoding revealed consistent accuracy in morphology-based identification of specimens to G. marginata s.l. complex. Prompt and careful morphological identification of ingested G. marginata s.l. has the potential to improve patient outcomes by leading to fast and appropriate treatment.; Methods

The dataset consists of concatenated sequences for 437 taxa, from the internal transcribed spacer DNA barcode region, the nuclear large subunit ribosomal RNA (LSU) gene and the RNA polymerase II second largest subunit gene (RPB2). The tree is a maximum likelihood tree from 200 search replicates with RAxML.

Taxon names are in the format:-

'G sideroides F25207 CA -TOX' where 'G sideroides' is a species in Galerina; F25207 is the herbarium accession number; CA is the two-digit code for country of collection; and -TOX indicates that the specimen was assayed and lacked detectable amatoxins.

+TOX would mean that the specimen was assayed and contained detectable amatoxins.

; Usage notes

This is a nexus file created in Mesquite.

Item Metadata

Title	Phylogenetic analysis of the distribution of deadly amatoxins among the little brown mushrooms of the genus Galerina
Creator	Berbee, Mary; Landry, Brandon; Whitton, Jeannette; Bazzicalupo, Anna; Ceska, Oldriska
Date Issued	2021-05-20
Description	<b>Abstract</b><br/> Some but not all of the species of 'little brown mushrooms' in the genus <i>Galerina</i> contain deadly amatoxins at concentrations equaling those in the death cap, <i>Amanita</i> <i>phalloides.</i> However, <i>Galerina</i>'s ~300 species are notoriously difficult to identify by morphology and the identity of toxin-containing specimens has not been verified with DNA barcode sequencing. This left open the question of which <i>Galerina</i> species contain toxins and which do not. We selected specimens for toxin analysis using a preliminary phylogeny of the fungal DNA barcode region, the ribosomal internal transcribed spacer (ITS) region. Using liquid chromatography/mass spectrometry, we analyzed amatoxins from 70 samples of <i>Galerina</i> and close relatives, collected in western British Columbia, Canada. To put the presence of toxins into a phylogenetic context, we included the 70 samples in maximum likelihood analyses of 438 taxa, using ITS, RNA polymerase II second largest subunit gene (<i>RPB2</i>), and nuclear large subunit ribosomal RNA (LSU) gene sequences. We sequenced barcode DNA from types where possible to aid with applications of names. We detected amatoxins only in the 24 samples of the <i>G. marginata</i> s.l. complex in the <i>Naucoriopsis</i> clade. We delimited 56 putative <i>Galerina</i> species using Automatic Barcode Gap Detection software. Phylogenetic analysis showed moderate to strong support for <i>Galerina</i> infrageneric clades <i>Naucoriopsis</i>, <i>Galerina</i>, <i>Tubariopsis</i>, and<i> Sideroides</i>. <i>Mycenopsis </i>appeared paraphyletic and included <i>Gymnopilus</i>. Amatoxins were not detected in 46 samples from <i>Galerina</i> clades outside of <i>Naucoriopsis</i> or from outgroups. Our data show significant quantities of toxin in all mushrooms tested from the <i>G. marginata</i> s.l. complex. DNA barcoding revealed consistent accuracy in morphology-based identification of specimens to <i>G.</i> <i>marginata </i>s.l. complex. Prompt and careful morphological identification of ingested <i>G.</i> <i>marginata </i>s.l. has the potential to improve patient outcomes by leading to fast and appropriate treatment.</p>; <b>Methods</b><br /> The dataset consists of concatenated sequences for 437 taxa, from the internal transcribed spacer DNA barcode region, the nuclear large subunit ribosomal RNA (LSU) gene and the RNA polymerase II second largest subunit gene (<i>RPB2</i>). The tree is a maximum likelihood tree from 200 search replicates with RAxML.</p> </p> Taxon names are in the format:-</p> 'G sideroides F25207 CA -TOX' where 'G sideroides' is a species in Galerina; F25207 is the herbarium accession number; CA is the two-digit code for country of collection; and -TOX indicates that the specimen was assayed and lacked detectable amatoxins.</p> +TOX would mean that the specimen was assayed and contained detectable amatoxins.</p> </p>; <b>Usage notes</b><br /> This is a nexus file created in Mesquite. </p>
Subject	Other
Type	Dataset
Notes	Dryad version number: 3</p> Version status: submitted</p> Dryad curation status: Published</p> Sharing link: https://datadryad.org/stash/share/98cfic5OnQlBkaCGyaPeLnT7voKC9dYdwjSAqbkAov8</p> Storage size: 3632548</p> Visibility: public</p>
Date Available	2020-11-30
Provider	University of British Columbia Library
License	CC0 1.0
DOI	10.14288/1.0397945
URI	https://doi.org/10.5683/SP2/U6KCQD
Publisher DOI	https://doi.org/10.5683/SP2/U6KCQD; https://doi.org/10.5061/dryad.r7sqv9s9z
Grant Funding Agency	Natural Sciences and Engineering Research Council of Canada
Rights URI	http://creativecommons.org/publicdomain/zero/1.0
Aggregated Source Repository	Dataverse

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