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Karl-Henrik Larsson

Publications and source records attributed to Karl-Henrik Larsson.

9 recordsLinked to original sources

Taxonomic reliability of DNA sequences in public sequence databases: a fungal perspective.

BACKGROUND: DNA sequences are increasingly seen as one of the primary information sources for species identification in many organism groups. Such approaches, popularly known as barcoding, are underpinned by the assumption that the reference databases used for comparison are sufficiently complete and feature correctly and informatively annotated entries. METHODOLOGY/PRINCIPAL FINDINGS: The present study uses a large set of fungal DNA sequences from the inclusive International Nucleotide Sequence Database to show that the taxon sampling of fungi is far from complete, that about 20% of the entries may be incorrectly identified to species level, and that the majority of entries lack descriptive and up-to-date annotations. CONCLUSIONS: The problems with taxonomic reliability and insufficient annotations in public DNA repositories form a tangible obstacle to sequence-based species identification, and it is manifest that the greatest challenges to biological barcoding will be of taxonomical, rather than technical, nature.

DNA, Fungal↗

Molecular phylogeny of Hyphoderma and the reinstatement of Peniophorella.

Hyphoderma is a large genus of corticioid homobasidiomycetes. In earlier homobasidiomycete-wide molecular phylogenetic studies the genus has appeared as polyphyletic. This paper describes the results from phylogenetic analyses of 22 species of Hyphoderma using nuclear 5.8 S and 28 S rDNA. Species with echinocysts and stephanocysts form a distinct clade well separated from Hyphoderma s. str. For this group the old genus name Peniophorella is available with P. pubera as the type species. Nineteen new combinations in Peniophorella are made and a key to the species is given. The clade representing Hyphoderma in its restricted sense receives only indicative support and a further subdivision of the genus may become necessary. H. capitatum, H. orphanellum, and H. sibiricum belong neither to Peniophorella nor to Hyphoderma s. str. Hypochnicium is a sister group to Hyphoderma. The phylogenetic analyses support the segregation of Hypochnicium analogum and H. vellereum as Gloeohypochnicium analogum and Granulobasidium vellereum, respectively, and the inclusion of H. detriticum in Hyphodontia. Hyphodermopsis and Bulbillomyces are best regarded as synonyms of Hypochnicium.

Base Sequence↗

Fruiting body-guided molecular identification of root-tip mantle mycelia provides strong indications of ectomycorrhizal associations in two species of Sistotrema (Basidiomycota).

Fruiting body guided sequence analysis of mycorrhizal root-tip mycelia is a powerful yet relatively sparsely explored method for species-level identification of mycorrhizal fungi. It is used in this study to indicate mycorrhizal associations in the corticioid (resupinate) genus Sistotrema of the cantharelloid clade through phylogenetic analysis of the ITS and nuLSU rDNA regions of two spatiotemporally co-occurring Sistotrema fruiting bodies and ectomycorrhizal root tips. The genus Sistotrema is confirmed to be polyphyletic, and the mycorrhizal species form a strongly supported monophyletic clade together with the stipitate genus Hydnum. The remaining lineages of Sistotrema may well be saprotrophic, the nutritional mode traditionally attributed to the genus, but the phylogenetic analyses show that they should be excluded from Sistotrema. The cantharelloid clade contains several mycorrhizal genera, but no symbiotic associations have previously been demonstrated for Sistotrema.

Alnus↗

Approaching the taxonomic affiliation of unidentified sequences in public databases--an example from the mycorrhizal fungi.

BACKGROUND: During the last few years, DNA sequence analysis has become one of the primary means of taxonomic identification of species, particularly so for species that are minute or otherwise lack distinct, readily obtainable morphological characters. Although the number of sequences available for comparison in public databases such as GenBank increases exponentially, only a minuscule fraction of all organisms have been sequenced, leaving taxon sampling a momentous problem for sequence-based taxonomic identification. When querying GenBank with a set of unidentified sequences, a considerable proportion typically lack fully identified matches, forming an ever-mounting pile of sequences that the researcher will have to monitor manually in the hope that new, clarifying sequences have been submitted by other researchers. To alleviate these concerns, a project to automatically monitor select unidentified sequences in GenBank for taxonomic progress through repeated local BLAST searches was initiated. Mycorrhizal fungi--a field where species identification often is prohibitively complex--and the much used ITS locus were chosen as test bed. RESULTS: A Perl script package called emerencia is presented. On a regular basis, it downloads select sequences from GenBank, separates the identified sequences from those insufficiently identified, and performs BLAST searches between these two datasets, storing all results in an SQL database. On the accompanying web-service http://emerencia.math.chalmers.se, users can monitor the taxonomic progress of insufficiently identified sequences over time, either through active searches or by signing up for e-mail notification upon disclosure of better matches. Other search categories, such as listing all insufficiently identified sequences (and their present best fully identified matches) publication-wise, are also available. DISCUSSION: The ever-increasing use of DNA sequences for identification purposes largely falls back on the assumption that public sequence databases contain a thorough sampling of taxonomically well-annotated sequences. Taxonomy, held by some to be an old-fashioned trade, has accordingly never been more important. emerencia does not automate the taxonomic process, but it does allow researchers to focus their efforts elsewhere than countless manual BLAST runs and arduous sieving of BLAST hit lists. The emerencia system is available on an open source basis for local installation with any organism and gene group as targets.

Classification↗

Airborne fungal colonisation of coarse woody debris in North Temperate Picea abies forest: impact of season and local spatial scale.

Coarse woody debris is important for mycodiversity in forest ecosystems, but its availability in managed stands is reduced. Leaving dead wood during felling is suggested as an option to sustain and restore the diversity. However, little is known what fungi would colonise freshly cut wood left on managed sites, and how the colonisation process is influenced by ecological factors. During summer and autumn, 120 freshly cut Picea abies stem sections over 8 cm in diameter were placed upright in mapped locations over two discrete plots separated by 100 m in a north-temperate forest. After seven weeks the sections were collected, and isolation and identification of fungi was done from their upper surfaces. In all 943 fungal strains were isolated, representing 97 species. Species richness in the summer survey was 42.5% higher than during the autumn survey. Low species similarity characterized the different seasons (Sorensen indices: S(S) = 0.36 and S(N) = 0.34) and for 21 species (22%) observation frequency was significantly affected by season. As a result, community structures in summer and autumn differed notably (z-test; P < 0.001). Species richness between the two plots differed by less than 10%, but there were 64 species (66%) found only in one of them, thus qualitative similarity was low (S(S) = 0.49). Quantitative similarity was higher (S(N) = 0.63), indicating that the dominant species colonised wood to a similar extent in both areas. Fungal community structure differed significantly among the two plots (z-test; P < 0.001). Our data showed that freshly cut CWD contributed to mycodiversity in managed north-temperate forest, providing habitats for numerous individuals from over 100 species. The fungal community within a single stand differed markedly both across small distances and over the seasons. In order to sustain and enhance mycodiversity in managed stands. coarse wood should always be left during harvesting. This study also demonstrates the importance of molecular identification and ITS sequence databases for exploring fungal diversity in natural communities.

Air Microbiology↗

UNITE: a database providing web-based methods for the molecular identification of ectomycorrhizal fungi.

Identification of ectomycorrhizal (ECM) fungi is often achieved through comparisons of ribosomal DNA internal transcribed spacer (ITS) sequences with accessioned sequences deposited in public databases. A major problem encountered is that annotation of the sequences in these databases is not always complete or trustworthy. In order to overcome this deficiency, we report on UNITE, an open-access database. UNITE comprises well annotated fungal ITS sequences from well defined herbarium specimens that include full herbarium reference identification data, collector/source and ecological data. At present UNITE contains 758 ITS sequences from 455 species and 67 genera of ECM fungi. UNITE can be searched by taxon name, via sequence similarity using blastn, and via phylogenetic sequence identification using galaxie. Following implementation, galaxie performs a phylogenetic analysis of the query sequence after alignment either to pre-existing generic alignments, or to matches retrieved from a blast search on the UNITE data. It should be noted that the current version of UNITE is dedicated to the reliable identification of ECM fungi. The UNITE database is accessible through the URL http://unite.zbi.ee

DNA, Ribosomal Spacer↗

galaxieEST: addressing EST identity through automated phylogenetic analysis.

BACKGROUND: Research involving expressed sequence tags (ESTs) is intricately coupled to the existence of large, well-annotated sequence repositories. Comparatively complete and satisfactory annotated public sequence libraries are, however, available only for a limited range of organisms, rendering the absence of sequences and gene structure information a tangible problem for those working with taxa lacking an EST or genome sequencing project. Paralogous genes belonging to the same gene family but distinguished by derived characteristics are particularly prone to misidentification and erroneous annotation; high but incomplete levels of sequence similarity are typically difficult to interpret and have formed the basis of many unsubstantiated assumptions of orthology. In these cases, a phylogenetic study of the query sequence together with the most similar sequences in the database may be of great value to the identification process. In order to facilitate this laborious procedure, a project to employ automated phylogenetic analysis in the identification of ESTs was initiated. RESULTS: galaxieEST is an open source Perl-CGI script package designed to complement traditional similarity-based identification of EST sequences through employment of automated phylogenetic analysis. It uses a series of BLAST runs as a sieve to retrieve nucleotide and protein sequences for inclusion in neighbour joining and parsimony analyses; the output includes the BLAST output, the results of the phylogenetic analyses, and the corresponding multiple alignments. galaxieEST is available as an on-line web service for identification of fungal ESTs and for download / local installation for use with any organism group at http://galaxie.cgb.ki.se/galaxieEST.html. CONCLUSIONS: By addressing sequence relatedness in addition to similarity, galaxieEST provides an integrative view on EST origin and identity, which may prove particularly useful in cases where similarity searches return one or more pertinent, but not full, matches and additional information on the query EST is needed.

Computational Biology↗

galaxie--CGI scripts for sequence identification through automated phylogenetic analysis.

MOTIVATION: The prevalent use of similarity searches like BLAST to identify sequences and species implicitly assumes the reference database to be of extensive sequence sampling. This is often not the case, restraining the correctness of the outcome as a basis for sequence identification. Phylogenetic inference outperforms similarity searches in retrieving correct phylogenies and consequently sequence identities, and a project was initiated to design a freely available script package for sequence identification through automated Web-based phylogenetic analysis. RESULTS: Three CGI scripts were designed to facilitate qualified sequence identification from a Web interface. Query sequences are aligned to pre-made alignments or to alignments made by ClustalW with entries retrieved from a BLAST search. The subsequent phylogenetic analysis is based on the PHYLIP package for inferring neighbor-joining and parsimony trees. The scripts are highly configurable. AVAILABILITY: A service installation and a version for local use are found at http://andromeda.botany.gu.se/galaxiewelcome.html and http://galaxie.cgb.ki.se

Algorithms↗

High phylogenetic diversity among corticioid homobasidiomycetes.

Homobasidiomycetes display a variety of fruit body morphologies. Examples include gilled mushrooms, coral fungi, polypores and puffballs but also species with simple crust-like basidiomata, usually called corticioid fungi. The latter group has largely been neglected in recent studies of homobasidiomycete evolution. The major goal of the present study was to explore the impact that the addition of a wide selection of species with crust-like basidiomata would have on homobasidiomycete phylogeny. Two genes, 5.8S and 28S in the nuclear rDNA repeats, were sequenced and a data set with 178 taxa analysed using neighbour-joining and maximum parsimony methods. Support for clades was evaluated by bootstrap. Basal nodes generally received weak support and branching order for major clades remained largely unresolved. Twelve major groups were recovered and corticioid fungi make up a major or important constituent in most of them. Nine groups are strongly supported but support for euagarics and polyporoid clades is poor. Phlebioid fungi were in earlier studies merged with the polyporoid clade but are here identified as a separate clade. Athelia is allied with ectomycorrhizal genera, inter alia Piloderma and Amphinema, in a separate clade forming a sister group to the boletes. We conclude that corticioid fungi hold a considerable share of the phylogenetic diversity displayed by homobasidiomycetes, and should always be considered when phylogenetic studies of larger basidiomycetes are designed.

Basidiomycota↗