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Native New Zealand forget-me-nots (Myosotis, Boraginaceae) comprise a Pleistocene species radiation with very low genetic divergence

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Abstract

Reconstruction of molecular phylogenies is an important step towards understanding the evolutionary history of island plant radiations. The New Zealand forget-me-nots (Myosotis, Boraginaceae) comprise a lineage of over 40 closely related but morphologically and ecologically diverse species whose evolutionary history and taxonomy are unclear. Myosotis is a high priority for systematic research in New Zealand because a high proportion of these species are threatened, and many have restricted geographic ranges and occupy very specific habitats. Here, we investigated the relationships and age of Southern Hemisphere forget-me-nots by performing phylogenetic, molecular dating, and other analyses of DNA sequence datasets from representatives of nearly all described species. To this end, we used both chloroplast (atpI–atpH + rps16–trnQ) and nuclear ribosomal (ITS + ETS) DNA sequences, as well as amplified fragment length polymorphisms (AFLPs). Our analyses showed that genus Myosotis likely arose in the Northern Hemisphere during the Miocene with the ancestor of the Southern Hemisphere lineage arising in the Pleistocene and radiating shortly thereafter. The Southern Hemisphere Myosotis species have very low levels of genetic divergence and their relationships are largely unresolved, likely due to a combination of recent radiation, hybridization, and incomplete lineage sorting. Our results are compared to those of similar studies on other New Zealand species radiations, and implications for ongoing and future Myosotis taxonomic and evolutionary research are discussed.

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Acknowledgments

We would like to thank Pat Brownsey and Ilse Breitwieser for their support of this project, staff from AK, CHR and WELT, the Department of Conservation (DOC) for permission and help to collect samples on DOC land, the many colleagues who helped with field work and sample collection (especially John Barkla, Andrea Brandon, Fiona Cameron, Hamish Carson, Bill Clarkson, Shannel Courtney, Pat Enright, Phil Garnock-Jones, David Glenny, Ant Kusabs, Alan Lee, Mike Lusk, Viv McGlynn, Brian Rance, Nick Singers, Jorge Santos, Barry Sneddon, Bec Stanley, Mike Thorsen, Steve Wagstaff, and Andreas Zeller), Dirk Albach, Dick Olmstead, and Matt Renner for sending Boraginaceae outgroup tissue and DNA, Peter Ritchie for use of the Molecular Ecology lab at Victoria University of Wellington to generate the DNA sequence and AFLP data, Simon Pfanzelt for key discussions and unwavering help running BEAST analyses, Phil Garnock-Jones for commenting on an earlier version of the manuscript, and two anonymous reviewers for their highly constructive criticisms during the review process. We acknowledge use of information contained in the New Zealand Fossil Record File. This research was supported by Core funding for Crown Research Institutes from the Ministry of Business, Innovation and Employment’s Science and Innovation Group.

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  1. Museum of New Zealand Te Papa Tongarewa, PO Box 467, Cable St, Wellington, 6140, New Zealand

    Heidi M. Meudt, Jessica M. Prebble & Carlos A. Lehnebach

  2. School of Biological Sciences, Victoria University of Wellington, Wellington, 6140, New Zealand

    Heidi M. Meudt & Carlos A. Lehnebach

  3. Institute of Fundamental Sciences, Massey University, Private Bag 11222, Palmerston North, 4442, New Zealand

    Jessica M. Prebble

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  1. Heidi M. Meudt
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Correspondence to Heidi M. Meudt.

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606_2014_1166_MOESM1_ESM.pdf

Online Resource 1 Table showing voucher details including GenBank accession numbers. ITS sequences downloaded from GenBank include a reference; all other sequences were newly sequenced for this study. Author names are given for species, subspecies and varieties not listed in Table 1 (PDF 150 kb)

606_2014_1166_MOESM2_ESM.pdf

Online Resource 2 RAxML phylogram of combined nuclear (ITS + ETS) DNA sequence dataset; scale units are substitutions per site. Individual names are genus + species epithet + Genbank number for the atpI–atpH sequence (or if no atpI–atpH sequence, then Genbank number for the ITS sequence). Numbers above branches are bootstrap values from 1000 replicates. L, C and M indicate the location of the calibration points based on Lithospermum, Cryptantha and New Zealand Myosotis fossils respectively (note these are shown here for illustrative purposes only as they were used in the BEAST analyses not in this RAxML anlaysis; see text for details). Voucher information for all individuals is given in Online Resource 1 (PDF 978 kb)

606_2014_1166_MOESM3_ESM.pdf

Online Resource 3 RAxML phylogram of combined chloroplast DNA (rps16-trnQ+atpI–atpH intergenic spacer regions) dataset; scale units are substitutions per site. Individual names are genus + species epithet + Genbank number for the atpI–atpH sequence (or if no atpI–atpH sequence, then Genbank number for the ITS sequence). Numbers above branches are bootstrap values from 1000 replicates. Voucher information for all individuals is given in Online Resource 1 (PDF 526 kb)

606_2014_1166_MOESM4_ESM.pdf

Online Resource 4 BEAST chronogram of combined nuclear (ITS + ETS) DNA sequence dataset using Lithospermum (L) and Cryptantha (C) fossil calibration points with uniform priors (the Myosotis fossil calibration point [M] was not used in this analysis; see text for details). The scale line is in millions of years before present. Individual names are genus + species epithet + Genbank number for the atpI–atpH sequence (or if no atpI–atpH sequence, then Genbank number for the ITS sequence). Dark circles signify branches with posterior probability values >0.95. Voucher information for all individuals is given in Online Resource 1 (PDF 49 kb)

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Meudt, H.M., Prebble, J.M. & Lehnebach, C.A. Native New Zealand forget-me-nots (Myosotis, Boraginaceae) comprise a Pleistocene species radiation with very low genetic divergence. Plant Syst Evol 301, 1455–1471 (2015). https://doi.org/10.1007/s00606-014-1166-x

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