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Scheunert, Agnes ; Lautenschlager, Ulrich ; Ott, Tankred ; Oberprieler, Christoph

Nano‐Strainer: A workflow for the identification of single‐copy nuclear loci for plant systematic studies, using target capture kits and Oxford Nanopore long reads

Scheunert, Agnes , Lautenschlager, Ulrich , Ott, Tankred und Oberprieler, Christoph (2023) Nano‐Strainer: A workflow for the identification of single‐copy nuclear loci for plant systematic studies, using target capture kits and Oxford Nanopore long reads. Ecology and Evolution 13 (7).

Veröffentlichungsdatum dieses Volltextes: 20 Okt 2023 14:17
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.54888


Zusammenfassung

In modern plant systematics, target enrichment enables simultaneous analysis of hundreds of genes. However, when dealing with reticulate or polyploidization histories, few markers may suffice, but often are required to be single-copy, a condition that is not necessarily met with commercial capture kits. Also, large genome sizes can render target capture ineffective, so that amplicon sequencing ...

In modern plant systematics, target enrichment enables simultaneous analysis of hundreds of genes. However, when dealing with reticulate or polyploidization histories, few markers may suffice, but often are required to be single-copy, a condition that is not necessarily met with commercial capture kits. Also, large genome sizes can render target capture ineffective, so that amplicon sequencing would be preferable; however, knowledge about suitable loci is often missing. Here, we present a comprehensive workflow for the identification of putative single-copy nuclear markers in a genus of interest, by mining a small dataset from target capture using a few representative taxa. The proposed pipeline assesses sequence variability contained in the data from targeted loci and assigns reads to their respective genes, via a combined BLAST/clustering procedure. Cluster consensus sequences are then examined based on four pre-defined criteria presumably indicative for absence of paralogy. This is done by calculating four specialized indices; loci are ranked according to their performance in these indices, and top-scoring loci are considered putatively single- or low copy. The approach can be applied to any probe set. As it relies on long reads, the present contribution also provides template workflows for processing Nanopore-based target capture data. Obtained markers are further tested and then entered into amplicon sequencing. For the detection of possibly remaining paralogy in these data, which might occur in groups with rampant paralogy, we also employ the long-read assembly tool Canu. In diploid representatives of the young Compositae genus Leucanthemum, characterized by high levels of polyploidy, our approach resulted in successful amplification of 13 loci. Modifications to remove traces of paralogy were made in seven of these. A species tree from the markers correctly reproduced main relationships in the genus, however, at low resolution. The presented workflow has the potential to valuably support phylogenetic research, for example in polyploid plant groups.



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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftEcology and Evolution
Verlag:WILEY
Ort der Veröffentlichung:HOBOKEN
Band:13
Nummer des Zeitschriftenheftes oder des Kapitels:7
Datum18 Juli 2023
InstitutionenBiologie und Vorklinische Medizin > Institut für Pflanzenwissenschaften > Arbeitsgruppe Evolution und Systematik der Pflanzen (Prof. Dr. Christoph Oberprieler)
Identifikationsnummer
WertTyp
10.1002/ece3.10190DOI
Stichwörter / KeywordsLEUCANTHEMUM MILL. COMPOSITAE; PLURIFLORUM CLAN COMPOSITAE; SPECIES TREE RECONSTRUCTION; SEQUENCE; ANTHEMIDEAE; INFERENCE; ALIGNMENT; ASTERACEAE; ORTHOLOGY; CompCOS loci; Leucanthemum; polyploidization; probe kit; reticulation
Dewey-Dezimal-Klassifikation500 Naturwissenschaften und Mathematik > 580 Pflanzen (Botanik)
StatusVeröffentlicht
BegutachtetJa, diese Version wurde begutachtet
An der Universität Regensburg entstandenJa
URN der UB Regensburgurn:nbn:de:bvb:355-epub-548889
Dokumenten-ID54888

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