Eukaryotes in the Silva database
Laura Wegener Parfrey Knight lab, CU Boulder
Pelin Yilmaz – MPI Bremen Silva in general
• SSU and LSU rDNA only • Curated based on alignment, sequence quality metrics and length (reference sets greater than 1200 bp) Silva 108 • Qiime forma ed files currently available (from Qiime.org resources) • Tony Walters implemented 18S tutorial
Problems with Silva 108
• About 1/5 listed as uncultured eukaryote • Taxonomy based on NCBI • Not standardized for computa onal analyses – Variable numbers of levels/ranks – Tony generated RDP forma ed taxa maps – but just grab first categories Eukaryo c Taxonomy Working Group
• Collabora on between Silva ribosomal database (Pelin Yilmaz), ISOP systema cs commi ee and others with computa onal or taxonomic exper se. • Goals for the revised classifica on – reflect phylogeny – Interface with computa onal tools
Pelin Yilmaz and Frank Oliver Glockner Eukaryo c Taxonomy Working Group
• Revised classifica on based on ISOP taxonomy (Adl et al 2005 and updates) – Implemented in the Silva 111 release • Other databases (Greengenes and RDP) also commi ed to incorpora ng eukaryotes in next release. Eukaryo c Taxonomy Working Group
• Collabora on between Silva ribosomal database (Pelin Yilmaz), ISOP systema cs commi ee and others with computa onal or taxonomic exper se. • Goals for the revised classifica on – reflect phylogeny – Interface with computa onal tools Silva 111
• 71787 eukaryo c sequences • 13582 fungal sequences • Taxonomy assigned to uncultured sequences according to tree (e.g. uncultured ciliate rather than uncultured eukaryote) Silva 111 – taxonomy sources
• Deep eukaryotes and pro sts – ISOP (Interna onal Society of Pro stologists) classifica on Silva 111 – taxonomy sources Summary
451 taxa, 72 lineages
16 genes
Parfrey et al. 2010 Silva 111 – taxonomy sources
• Deep eukaryotes and pro sts - ISOP classifica on • Fungi – originally Pelin using Index Fungorum – Currently looking into Mycobank – Ideal: adopt same taxonomy as ITS database • Long term: aim to sync taxonomy with other efforts, e.g. Open tree of life project Silva 111 – taxonomy
• Also released flat file of taxonomy • Encourage users to use taxonomy of choice • Customize ranks (for RDP classifier) based on study Silva 108 curated tree
Animals
Fungi
Amoebozoa Excavata
Alveolates
Stramenopiles
Red algae Topiary Explorer Visualizing the high-throughput sequencing data within a phylogene c context
Developer: Meg Pirrung in Knight lab h p://topiaryexplorer.sourceforge.net/
A) Eukaryotic tree colored by taxonomy Environmental sequences B) Eukaryotic tree colored environment Rhizaria colored by clade
Stramenopiles Blastocystis
Ciliates
Green algae
C) Bacterial tree colored by environment Enterobacteria
Proteobacteria
Candida Fungi important in Fungi Tenericutes most environments. Actinobacteria Bi!dobacteria Prevotella Bacteroidetes
Acidobacteria
Animals Firmicutes
Closteridia
Entamoeba Amoebozoa Lactobacillus
Parabasalids Archaea Example of ranks file
Path node_name rank_name level Eukaryota Eukaryota domain Level 1 Eukaryota;Amb-18S-6341 Amb-18S-6341 phylum Level 2 Eukaryota;Amoebozoa Amoebozoa kingdom Level 2 Eukaryota;Amoebozoa;Archamoebae Archamoebae phylum Level 3 Eukaryota;Amoebozoa;Archamoebae;Entamoebida Entamoebida order Level 4 Eukaryota;Amoebozoa;Archamoebae;Entamoebida;Entamoeba Entamoeba genus Level 5 Eukaryota;Amoebozoa;Archamoebae;Mastigamoebaea Mastigamoebaea order Level 4 Eukaryota;Amoebozoa;Archamoebae;Mastigamoebaea;Mastiga moeba Mastigamoeba genus Level 5 Eukaryota;Amoebozoa;Cavosteliida Cavosteliida class Level 3 Eukaryota;Amoebozoa;Cavosteliida;Cavostelium Cavostelium genus Level 4 Eukaryota;Amoebozoa;Cavosteliida;MPE1-14 MPE1-14 order Level 4 Schizoplasmodio Eukaryota;Amoebozoa;Cavosteliida;Schizoplasmodiopsis psis genus Level 4 Eukaryota;Amoebozoa;Cavosteliida;Tychosporium Tychosporium genus Level 4 Eukaryota;Amoebozoa;Dictyostelia Dictyostelia class Level 3 Eukaryota;Amoebozoa;Dictyostelia;Acytostelium Acytostelium genus Level 4 Eukaryota;Amoebozoa;Dictyostelia;Dictyostelium Dictyostelium genus Level 4 Eukaryota;Amoebozoa;Dictyostelia;Polysphondylium Polysphondylium genus Level 4 Eukaryota;Amoebozoa;Discosea Discosea phylum Level 3 Eukaryota;Amoebozoa;Discosea;Flabellinia Flabellinia class Level 4
Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida Dactylopodida order Level 5 Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida;A1W VB A1WVB family Level 6 Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida;Koro tnevella Korotnevella genus Level 6 Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida;Neo paramoeba Neoparamoeba genus Level 6 Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida;Pse Pseudoparamoe udoparamoeba ba genus Level 6 Eukaryota;Amoebozoa;Discosea;Flabellinia;Dactylopodida;Vexil lifera Vexillifera genus Level 6 Release of Qiime forma ed files
• Later this fall • 99% and 97% representa ve sequences – Aligned and unaligned – Genbank accession is the iden fier • Taxa map – Full – RDP forma ed • Tree for eukaryotes only How to integrate ITS and 18S based classifica on / databases?
Collaborators: Rob Knight (CU) Acknowledgements Pelin Yilmaz (Silva) Frank Oliver Glockner (Silva)
Knight Lab: Tony Walters Matt Gebert Chris Lauber Daniel McDonald Jose C Clemente Scott Bates Jessica Metcalf
Eukaryo c taxonomy working group