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Scheunert, Agnes ; Dorfner, Marco ; Lingl, Thomas ; Oberprieler, Christoph

Can we use it? On the utility of de novo and reference-based assembly of Nanopore data for plant plastome sequencing

Scheunert, Agnes, Dorfner, Marco, Lingl, Thomas und Oberprieler, Christoph (2020) Can we use it? On the utility of de novo and reference-based assembly of Nanopore data for plant plastome sequencing. PLOS ONE 15 (3), e0226234.

Veröffentlichungsdatum dieses Volltextes: 22 Apr 2020 15:17
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.43098


Zusammenfassung

The chloroplast genome harbors plenty of valuable information for phylogenetic research. Illumina short-read data is generally used for de novo assembly of whole plastomes. PacBio or Oxford Nanopore long reads are additionally employed in hybrid approaches to enable assembly across the highly similar inverted repeats of a chloroplast genome. Unlike for PacBio, plastome assemblies based solely on ...

The chloroplast genome harbors plenty of valuable information for phylogenetic research. Illumina short-read data is generally used for de novo assembly of whole plastomes. PacBio or Oxford Nanopore long reads are additionally employed in hybrid approaches to enable assembly across the highly similar inverted repeats of a chloroplast genome. Unlike for PacBio, plastome assemblies based solely on Nanopore reads are rarely found, due to their high error rate and non-random error profile. However, the actual quality decline connected to their use has rarely been quantified. Furthermore, no study has employed reference-based assembly using Nanopore reads, which is common with Illumina data. Using Leucanthemum Mill. as an example, we compared the sequence quality of seven chloroplast genome assemblies of the same species, using combinations of two sequencing platforms and three analysis pipelines. In addition, we assessed the factors which might influence Nanopore assembly quality during sequence generation and bioinformatic processing. The consensus sequence derived from de novo assembly of Nanopore data had a sequence identity of 99.59% compared to Illumina short-read de novo assembly. Most of the errors detected were indels (81.5%), and a large majority of them is part of homopolymer regions. The quality of reference-based assembly is heavily dependent upon the choice of a close-enough reference. When using a reference with 0.83% sequence divergence from the studied species, mapping of Nanopore reads results in a consensus comparable to that from Nanopore de novo assembly, and of only slightly inferior quality compared to a reference-based assembly with Illumina data. For optimal de novo assembly of Nanopore data, appropriate filtering of contaminants and chimeric sequences, as well as employing moderate read coverage, is essential. Based on these results, we conclude that Nanopore long reads are a suitable alternative to Illumina short reads in plastome phylogenomics. Few errors remain in the finalized assembly, which can be easily masked in phylogenetic analyses without loss in analytical accuracy. The easily applicable and cost-effective technology might warrant more attention by researchers dealing with plant chloroplast genomes.



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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftPLOS ONE
Verlag:PLOS
Ort der Veröffentlichung:SAN FRANCISCO
Band:15
Nummer des Zeitschriftenheftes oder des Kapitels:3
Seitenbereich:e0226234
Datum24 März 2020
InstitutionenBiologie und Vorklinische Medizin > Institut für Pflanzenwissenschaften > Arbeitsgruppe Evolution und Systematik der Pflanzen (Prof. Dr. Christoph Oberprieler)
Biologie und Vorklinische Medizin > Institut für Pflanzenwissenschaften > Arbeitsgruppe Evolution und Systematik der Pflanzen (Prof. Dr. Christoph Oberprieler)
Identifikationsnummer
WertTyp
10.1371/journal.pone.0226234DOI
Stichwörter / KeywordsCHLOROPLAST DNA; READ ALIGNMENT; GENOME; MITOCHONDRIAL; EVOLUTIONARY; ANGIOSPERMS; POPULATION; REPEAT; REGION; RATES;
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-430986
Dokumenten-ID43098

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