Title
Nanopore sequencing of long ribosomal DNA amplicons enables portable and simple biodiversity assessments with high phylogenetic resolution across broad taxonomic scale
Date Issued
01 May 2019
Access level
open access
Resource Type
journal article
Author(s)
Krehenwinkel H.
Pomerantz A.
Henderson J.B.
Kennedy S.R.
Lim J.Y.
Swamy V.
Graham N.
Patel N.H.
Gillespie R.G.
Prost S.
Publisher(s)
Oxford University Press
Abstract
Background: In light of the current biodiversity crisis, DNA barcoding is developing into an essential tool to quantify state shifts in global ecosystems. Current barcoding protocols often rely on short amplicon sequences, which yield accurate identification of biological entities in a community but provide limited phylogenetic resolution across broad taxonomic scales. However, the phylogenetic structure of communities is an essential component of biodiversity. Consequently, a barcoding approach is required that unites robust taxonomic assignment power and high phylogenetic utility. A possible solution is offered by sequencing long ribosomal DNA (rDNA) amplicons on the MinION platform (Oxford Nanopore Technologies). Findings: Using a dataset of various animal and plant species, with a focus on arthropods, we assemble a pipeline for long rDNA barcode analysis and introduce a new software (MiniBar) to demultiplex dual indexed Nanopore reads. We find excellent phylogenetic and taxonomic resolution offered by long rDNA sequences across broad taxonomic scales. We highlight the simplicity of our approach by field barcoding with a miniaturized, mobile laboratory in a remote rainforest. We also test the utility of long rDNA amplicons for analysis of community diversity through metabarcoding and find that they recover highly skewed diversity estimates. Conclusions: Sequencing dual indexed, long rDNA amplicons on the MinION platform is a straightforward, cost-effective, portable, and universal approach for eukaryote DNA barcoding. Although bulk community analyses using long-amplicon approaches may introduce biases, the long rDNA amplicons approach signifies a powerful tool for enabling the accurate recovery of taxonomic and phylogenetic diversity across biological communities.
Volume
8
Issue
5
Language
English
OCDE Knowledge area
Conservación de la Biodiversidad Bioquímica, Biología molecular Nano-procesos
Scopus EID
2-s2.0-85065222871
PubMed ID
Source
GigaScience
ISSN of the container
2047217X
Sponsor(s)
We thank Taylor Liu for help during laboratory work, and Tara Gallant for help during specimen collection. Hitomi Asahara graciously provided access to a laboratory facility and the necessary software for our MinION sequencing run. We thank the State of Hawaii Department of Land and Natural Resources and the Ser-vicio Nacional Forestal y de Fauna Silvestre, who provided collection permits and rainforest expeditions, and Gabriela Orihuela for providing assistance and support with fieldwork in Peru. We thank Anna Holmquist for providing the Psechrus sp. specimen. The specimen was collected with permits from the Indonesian Ministry of Research and Technology (KEMENRISTEK) and in collaboration with Pungki Lupiyaningdyah and Anang Achmadi from the Museum Zoologicum Bogoriense and funded by a National Science Foundation grant DEB 1457845.
Sources of information: Directorio de Producción Científica Scopus