Abstract

The inference of evolutionary relationships in the species-rich family Orchidaceae has hitherto relied heavily on plastid DNA sequences and limited taxon sampling. Previous studies have provided a robust plastid phylogenetic framework, which was used to classify orchids and investigate the drivers of orchid diversification. However, the extent to which phylogenetic inference based on the plastid genome is congruent with the nuclear genome has been only poorly assessed. We inferred higher-level phylogenetic relationships of orchids based on likelihood and ASTRAL analyses of 294 low-copy nuclear genes sequenced using the Angiosperms353 universal probe set for 75 species (representing 69 genera, 16 tribes, 24 subtribes) and a concatenated analysis of 78 plastid genes for 264 species (117 genera, 18 tribes, 28 subtribes). We compared phylogenetic informativeness and support for the nuclear and plastid phylogenetic hypotheses. Phylogenetic inference using nuclear data sets provides well-supported orchid relationships that are highly congruent between analyses. Comparisons of nuclear gene trees and a plastid supermatrix tree showed that the trees are mostly congruent, but revealed instances of strongly supported phylogenetic incongruence in both shallow and deep time. The phylogenetic informativeness of individual Angiosperms353 genes is in general better than that of most plastid genes. Our study provides the first robust nuclear phylogenomic framework for Orchidaceae and an assessment of intragenomic nuclear discordance, plastid-nuclear tree incongruence, and phylogenetic informativeness across the family. Our results also demonstrate what has long been known but rarely thoroughly documented: nuclear and plastid phylogenetic trees can contain strongly supported discordances, and this incongruence must be reconciled prior to interpretation in evolutionary studies, such as taxonomy, biogeography, and character evolution.

Highlights

  • MethodsWe inferred higher-­level phylogenetic relationships of orchids based on likelihood and ASTRAL analyses of 294 low-­copy nuclear genes sequenced using the Angiosperms353 universal probe set for 75 species (representing 69 genera, 16 tribes, 24 subtribes) and a concatenated analysis of 78 plastid genes for 264 species (117 genera, 18 tribes, 28 subtribes)

  • PREMISE: The inference of evolutionary relationships in the species-­rich family Orchidaceae has hitherto relied heavily on plastid DNA sequences and limited taxon sampling

  • Our results demonstrate what has long been known but rarely thoroughly documented: nuclear and plastid phylogenetic trees can contain strongly supported discordances, and this incongruence must be reconciled prior to interpretation in evolutionary studies, such as taxonomy, biogeography, and character evolution

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Summary

Methods

We inferred higher-­level phylogenetic relationships of orchids based on likelihood and ASTRAL analyses of 294 low-­copy nuclear genes sequenced using the Angiosperms353 universal probe set for 75 species (representing 69 genera, 16 tribes, 24 subtribes) and a concatenated analysis of 78 plastid genes for 264 species (117 genera, 18 tribes, 28 subtribes). 14 species from non-o­ rchid monocot families were included as outgroups (Appendix S1). Generated nuclear DNA data were produced from 62 vouchered accessions stored in the DNA and tissue bank of the Royal Botanic Gardens, Kew (https://dnabank.science.kew.org/homepage.html). These DNA samples had been previously extracted from silica-­ dried leaves using a modified CTAB method (Doyle and Doyle, 1987). Nuclear short Illumina sequencing reads were data-m­ ined from the Sequence Read Archive (SRA) using the fastq-­dump software of the SRAtool-k­ it package (available at https://ncbi.github.io/sra-­tools/install_config.html) and the 1KP data repository (Wong et al, 2020; https://sites.google.com/a/ualberta.ca/onekp/) to expand our taxon sampling with 13 additional orchid species

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