Next-generation polyploid phylogenetics: rapid resolution of hybrid polyploid complexes using PacBio single-molecule sequencing.

dc.contributor.author

Rothfels, CJ

dc.contributor.author

Pryer, KM

dc.contributor.author

Li, F

dc.date.accessioned

2020-11-30T15:08:06Z

dc.date.available

2020-11-30T15:08:06Z

dc.date.issued

2017-01

dc.date.updated

2020-11-30T15:08:05Z

dc.description.abstract

Difficulties in generating nuclear data for polyploids have impeded phylogenetic study of these groups. We describe a high-throughput protocol and an associated bioinformatics pipeline (Pipeline for Untangling Reticulate Complexes (Purc)) that is able to generate these data quickly and conveniently, and demonstrate its efficacy on accessions from the fern family Cystopteridaceae. We conclude with a demonstration of the downstream utility of these data by inferring a multi-labeled species tree for a subset of our accessions. We amplified four c. 1-kb-long nuclear loci and sequenced them in a parallel-tagged amplicon sequencing approach using the PacBio platform. Purc infers the final sequences from the raw reads via an iterative approach that corrects PCR and sequencing errors and removes PCR-mediated recombinant sequences (chimeras). We generated data for all gene copies (homeologs, paralogs, and segregating alleles) present in each of three sets of 50 mostly polyploid accessions, for four loci, in three PacBio runs (one run per set). From the raw sequencing reads, Purc was able to accurately infer the underlying sequences. This approach makes it easy and economical to study the phylogenetics of polyploids, and, in conjunction with recent analytical advances, facilitates investigation of broad patterns of polyploid evolution.

dc.identifier.issn

1469-8137

dc.identifier.issn

1469-8137

dc.identifier.uri

https://hdl.handle.net/10161/21753

dc.language

eng

dc.publisher

Wiley

dc.relation.ispartof

The New phytologist

dc.relation.isversionof

10.1111/nph.14111

dc.subject

Sequence Analysis, DNA

dc.subject

Hybridization, Genetic

dc.subject

Computational Biology

dc.subject

Phylogeny

dc.subject

Recombination, Genetic

dc.subject

Base Sequence

dc.subject

Consensus Sequence

dc.subject

Polyploidy

dc.subject

Alleles

dc.subject

Databases, Genetic

dc.subject

Biological Evolution

dc.title

Next-generation polyploid phylogenetics: rapid resolution of hybrid polyploid complexes using PacBio single-molecule sequencing.

dc.type

Journal article

duke.contributor.orcid

Pryer, KM|0000-0002-9776-6736

pubs.begin-page

413

pubs.end-page

429

pubs.issue

1

pubs.organisational-group

Trinity College of Arts & Sciences

pubs.organisational-group

Biology

pubs.organisational-group

Duke Science & Society

pubs.organisational-group

Duke

pubs.organisational-group

Initiatives

pubs.organisational-group

Institutes and Provost's Academic Units

pubs.publication-status

Published

pubs.volume

213

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
Rothfels et al.NP.2107.pdf
Size:
823.33 KB
Format:
Adobe Portable Document Format
Description:
Published version