Evolution of Tandem Repeats Is Mirroring Post-Polyploid Cladogenesis in Heliophila (Brassicaceae)
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fpls-11-607893 December 30, 2020 Time: 16:36 # 1 ORIGINAL RESEARCH published: 12 January 2021 doi: 10.3389/fpls.2020.607893 Evolution of Tandem Repeats Is Mirroring Post-polyploid Cladogenesis in Heliophila (Brassicaceae) Mert Dogan1,2, Milan Pouch1,2, Terezie Mandáková1,3, Petra Hloušková1, Xinyi Guo1, Pieter Winter4, Zuzana Chumová5, Adriaan Van Niekerk6, Klaus Mummenhoff7, Ihsan A. Al-Shehbaz8, Ladislav Mucina6,9 and Martin A. Lysak1,2* 1 CEITEC, Masaryk University, Brno, Czechia, 2 NCBR, Faculty of Science, Masaryk University, Brno, Czechia, 3 Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czechia, 4 South African National Biodiversity Institute (SANBI), Kirstenbosch, Cape Town, South Africa, 5 Institute of Botany, Czech Academy of Sciences, Prùhonice, Czechia, 6 Department of Geography & Environmental Studies, Stellenbosch University, Stellenbosch, South Africa, 7 Department of Biology, Botany, Osnabrück University, Osnabrück, Germany, 8 Missouri Botanical Garden, St. Louis, MO, United States, 9 Harry Butler Institute, Murdoch University, Perth, WA, Australia Edited by: Christoph Oberprieler, The unigeneric tribe Heliophileae encompassing more than 100 Heliophila species University of Regensburg, Germany is morphologically the most diverse Brassicaceae lineage. The tribe is endemic to Reviewed by: J. Chris Pires, southern Africa, confined chiefly to the southwestern South Africa, home of two University of Missouri, United States biodiversity hotspots (Cape Floristic Region and Succulent Karoo). The monospecific Ales Kovarik, Academy of Sciences of the Czech Chamira (C. circaeoides), the only crucifer species with persistent cotyledons, Republic (ASCR), Czechia is traditionally retrieved as the closest relative of Heliophileae. Our transcriptome *Correspondence: analysis revealed a whole-genome duplication (WGD) ∼26.15–29.20 million years ago, Martin A. Lysak presumably preceding the Chamira/Heliophila split. The WGD was then followed by [email protected]; [email protected] genome-wide diploidization, species radiations, and cladogenesis in Heliophila. The expanded phylogeny based on nuclear ribosomal DNA internal transcribed spacer Specialty section: (ITS) uncovered four major infrageneric clades (A–D) in Heliophila and corroborated This article was submitted to Plant Systematics and Evolution, the sister relationship between Chamira and Heliophila. Herein, we analyzed how the a section of the journal diploidization process impacted the evolution of repetitive sequences through low- Frontiers in Plant Science coverage whole-genome sequencing of 15 Heliophila species, representing the four Received: 18 September 2020 Accepted: 16 November 2020 clades, and Chamira. Despite the firmly established infrageneric cladogenesis and Published: 12 January 2021 different ecological life histories (four perennials vs. 11 annual species), repeatome Citation: analysis showed overall comparable evolution of genome sizes (288–484 Mb) Dogan M, Pouch M, and repeat content (25.04–38.90%) across Heliophila species and clades. Among Mandáková T, Hloušková P, Guo X, Winter P, Chumová Z, Van Niekerk A, Heliophila species, long terminal repeat (LTR) retrotransposons were the predominant Mummenhoff K, Al-Shehbaz IA, components of the analyzed genomes (11.51–22.42%), whereas tandem repeats Mucina L and Lysak MA (2021) Evolution of Tandem Repeats Is had lower abundances (1.03–12.10%). In Chamira, the tandem repeat content Mirroring Post-polyploid Cladogenesis (17.92%, 16 diverse tandem repeats) equals the abundance of LTR retrotransposons in Heliophila (Brassicaceae). (16.69%). Among the 108 tandem repeats identified in Heliophila, only 16 repeats Front. Plant Sci. 11:607893. doi: 10.3389/fpls.2020.607893 were found to be shared among two or more species; no tandem repeats Frontiers in Plant Science | www.frontiersin.org 1 January 2021 | Volume 11 | Article 607893 fpls-11-607893 December 30, 2020 Time: 16:36 # 2 Dogan et al. Repeatome Evolution in Heliophileae were shared by Chamira and Heliophila genomes. Six “relic” tandem repeats were shared between any two different Heliophila clades by a common descent. Four and six clade-specific repeats shared among clade A and C species, respectively, support the monophyly of these two clades. Three repeats shared by all clade A species corroborate the recent diversification of this clade revealed by plastome-based molecular dating. Phylogenetic analysis based on repeat sequence similarities separated the Heliophila species to three clades [A, C, and (BCD)], mirroring the post-polyploid cladogenesis in Heliophila inferred from rDNA ITS and plastome sequences. Keywords: repetitive DNA, repeatome, whole-genome duplication (WGD), rDNA ITS, plastome phylogeny, Cruciferae, Cape flora, South Africa INTRODUCTION Despite Heliophila species being a frequent and sometimes dominating element of some southern African plant Geographically and phylogenetically well-defined groups are communities, there is limited knowledge of the phylogeographic ideal study objects to analyze the evolution of diverse genomic origin of the genus, interspecies relationships, and genome parameters during long periods of isolation that prevented evolution of Heliophila species. Mummenhoff et al. (2005) gene flow with other species groups. Although Brassicaceae published a pioneering study, laying foundations for follow-up (mustard family, Cruciferae) occur on all continents, except phylogenomic analyses, demonstrating monophyly of the tribe for Antarctica, and several weedy and crop species have a Heliophileae with South Africa’s endemic Chamira circaeoides worldwide distribution, some crucifer clades are restricted to as the sister species to Heliophila, finding support for rapid (sub)continents or smaller geographic regions (Lysak and Koch, diversification against a background of aridification in the 2011; Al-Shehbaz, 2012). For instance, tribes of the CES clade Pliocene/Pleistocene, and showing massive parallel evolution (i.e., Cremolobeae, Eudemeae, and Schizopetaleae), as well as of fruit characters traditionally used in the classification Halimolobeae, Physarieae, and all but one Thelypodieae species, of Heliophileae. Further, ecological optimization analysis are endemic to the New World, while Microlepidieae occur only allowed preliminary insights into the ecogeographical evolution in Australia and New Zealand. In Africa, the family has a reduced in Heliophileae. species and generic diversity, with the largest endemic clade The last phylogenetic study of c. 57 Heliophila species confined to southern Africa (South Africa, Lesotho, eSwatini, based on internal transcribed spacer (ITS) sequences suggested and Namibia). The tribe Heliophileae includes some 104 species basal polytomy involving three clades, all sister to Chamira (compilation by I. A. Al-Shehbaz) concentrated chiefly in the (Mandáková et al., 2012). The latter authors showed that two winter-rainfall region of the southwestern South Africa, home ITS clades are dominated by two chromosome numbers (2n = 20 to two global biodiversity hotspots – Cape Floristic Region and and 2n = 22), whereas the third clade mainly contained shrubby Succulent Karoo. Heliophila ranks among the largest crucifer species with chromosome numbers known only for two species genera, such as Alyssum, Boechera, Cardamine, Draba, Erysimum, at that time. Chromosome numbers in 27 analyzed Heliophila Lepidium, and Physaria (Al-Shehbaz, 2012). The genus is often species ranged from 2n = 16 to 2n = c. 88, presumably due regarded as morphologically the most diverse Brassicaceae to polyploidy and dysploidal chromosomal rearrangements. lineage (Mummenhoff et al., 2005). Heliophila varies from Interestingly, comparative chromosome painting analyses, small ephemeral annual to perennial herbs (incl. one lianella), revealing the duplicated nature of Heliophila genomes, suggested subshrubs, and tall shrubs (e.g., Heliophila brachycarpa). The the existence of an allohexaploid ancestor preceding the species vary particularly in foliage (entire to variously dissected); divergence of Heliophila lineages (Mandáková et al., 2012). petal length (1.2–30 mm) and color (white, pink, mauve, purple, This was supported by an analysis of synonymous substitution blue, or yellow); number and presence vs. absence of petal and rates (Ks) of paralogous and orthologous genes in Heliophila cf. stamen appendages; presence vs. absence of paired glands at the longifolia (Mandáková et al., 2017). bases of pedicels and/or leaves; ovule number (1–80); fruit length The high species diversity (>100 species) and extraordinary (2–120 mm long), shape (linear, lanceolate, oblong, ovate, elliptic, ecomorphological variability of Heliophila impacted by ancient orbicular), constriction (moniliform or not), type (silique, and more recent whole-genome duplication (WGD) events and silicle, samara, schizocarp), and flattening (terete, quadrangular, following post-polyploid diploidization (PPD), confined to one latiseptate, angustiseptate); gynophore length (obsolete to 12 mm of the most remarkable biodiversity hotspots, make the genus an long); style length (0.3–20 mm long) and shape (linear, intricate but attractive phylogenomic model. In this study, based filiform, conical, clavate, ovoid, globose); seed length (0.6–9 mm on the previous results and by including a broader spectrum long), shape, and development of wing; and cotyledonary type of species, we aim at providing new insight to the WGD– (diplecolobal, spirolobal) (Marais,