A brainstorm on the systematics of Turnera (Turneraceae, Malpighiales) caused by insights from molecular phylogenetics and morphological evolution.

With 145 species, Turnera is the largest genus of Turneraceae (Malpighiales). Despite several morphotaxonomic and cytogenetic studies, our knowledge about the phylogenetic relationships in Turnera remains mainly based on morphological data. Here, we reconstruct the most comprehensive phylogeny of Turnera with molecular data to understand the morphological evolution within this group and to assess its circumscription and infrageneric classification. We analyzed two nuclear and six plastid markers and 112 taxa, including species and infraspecific taxa, 97 from Turnera, covering the 11 series of the genus. Bayesian inference, maximum parsimony and maximum likelihood analyses show that Turnera, as traditionally circumscribed, is not monophyletic. The genus is divided into two well-supported independent clades; one of them is sister to the genus Piriqueta and is here segregated as the new genus Oxossia. According to our reconstructions, Turnera probably evolved from an ancestor without extrafloral nectaries and with solitary, homostylous flowers with yellow petals. The emergences of extrafloral nectaries and distyly, both common in extant taxa, played an important role in the diversification of the genus. An updated infrageneric classification reflecting the relationships within Turnera is now possible based on morphological synapomorphies and is here designed for further studies.

First report of laticifers in lianas of Malpighiaceae and their phylogenetic implications.

PREMISE: Laticifers have evolved multiple times in angiosperms and have been interpreted as a key innovation involved in plant defense mechanisms. In Malpighiaceae, laticifers were previously known from a single lineage of trees and shrubs, the Galphimia clade, but with detailed anatomical analyses here, we show that their distribution is broader in the family, also encompassing large clades of lianas. METHODS: From 15 genera, 70 species of Malpighiaceae were surveyed through careful anatomical ontogenetic analysis of roots, stems, and leaves and detailed histochemical tests to elucidate the nature of laticifers and latex in the family. RESULTS: Articulated anastomosing laticifers were encountered in roots, stems, and leaves of two distantly related megadiverse genera of Malpighiaceae lianas: Stigmaphyllon (stigmaphylloid clade) and Tetrapterys s.s. (tetrapteroid clade). From the apex downward, in Stigmaphyllon the laticifers are derived from the procambium and from the cambium during its early activity and are present in the outermost part of the vascular cylinder of stems and leaves and in the pericycle of roots, whereas in Tetrapterys s.s. they are derived from the ground meristem, procambium, and cambium throughout the plant body and are present in the cortex and pith, either the pericycle in roots or the outermost part of the vascular system in stems and leaves, and the primary and secondary phloem. CONCLUSIONS: Laticifers seem to have evolved at least three times independently in Malpighiaceae, once in a lineage of trees and shrubs and twice in two distantly related megadiverse lianescent lineages. Laticifer evolution in Malpighiaceae is homoplastic and may be related to increases in species diversification.

Dispersal largely explains the Gondwanan distribution of the ancient tropical clusioid plant clade.

PREMISE OF THE STUDY: The clusioid clade (Malpighiales) has an ancient fossil record (∼90 Ma) and extant representatives exhibit a pantropical distribution represented on all former Gondwanan landmasses (Africa, Australia, India, Madagascar, and South America) except Antarctica. Several biogeographers have hypothesized that the clusioid distribution is an example of Gondwanan vicariance. Our aim is to test the hypothesis that the modern distribution of the clusioid clade is largely explained by Gondwanan fragmentation. METHODS: Using a four gene, 207-taxon data set we simultaneously estimated the phylogeny and divergence times of the clusioid clade using a Bayesian Markov chain Monte Carlo approach. Ancestral Area Reconstructions (AARs) were then conducted on a distribution of 1000 trees and summarized on a reduced phylogeny. KEY RESULTS: Divergence time estimates and AARs revealed only two or four cladogenic events that are potentially consistent with Gondwanan vicariance, depending on the placement of the ancient fossil Paleoclusia. In contrast, dispersal occurred on > 25% of the branches, indicating the current distribution of the clade likely reflects extensive recent dispersal during the Cenozoic (< 65 Ma), most of which occurred after the beginning of the Eocene (∼56 Ma). CONCLUSIONS: These results support growing evidence that suggests many traditionally recognized angiosperm clades (families and genera) are too young for their distributions to have been influenced strictly by Gondwanan fragmentation. Instead, it appears that corridors of dispersal may be the best explanation for numerous angiosperm clades with Gondwanan distributions.

Understanding the spectacular failure of DNA barcoding in willows (Salix): does this result from a trans-specific selective sweep?

Willows (Salix: Salicaceae) form a major ecological component of Holarctic floras and consequently are an obvious target for a DNA-based identification system. We surveyed two to seven plastid genome regions (~3.8 kb; ~3% of the genome) from 71 Salix species across all five subgenera, to assess their performance as DNA barcode markers. Although Salix has a relatively high level of interspecific hybridization, this may not sufficiently explain the near complete failure of barcoding that we observed: only one species had a unique barcode. We recovered 39 unique haplotypes, from more than 500 specimens, that could be partitioned into six major haplotype groups. A unique variant of group I (haplotype 1*) was shared by 53 species in three of five Salix subgenera. This unusual pattern of haplotype sharing across infrageneric taxa is suggestive of either a massive nonrandom coalescence failure (incomplete lineage sorting), or of repeated plastid capture events, possibly including a historical selective sweep of haplotype 1* across taxonomic sections. The former is unlikely as molecular dating indicates that haplotype 1* originated recently and is nested in the oldest major haplotype group in the genus. Further, we detected significant non-neutrality in the frequency spectrum of mutations in group I, but not outside group I, and demonstrated a striking absence of geographical (isolation by distance) effects in the haplotype distributions of this group. The most likely explanation for the patterns we observed involves recent repeated plastid capture events, aided by widespread hybridization and long-range seed dispersal, but primarily propelled by one or more trans-species selective sweeps.

Phylogenetics, ancestral state reconstruction, and a new infrafamilial classification of the pantropical Ochnaceae (Medusagynaceae, Ochnaceae s.str., Quiinaceae) based on five DNA regions.

Ochnaceae s.str. (Malpighiales) are a pantropical family of about 500 species and 27 genera of almost exclusively woody plants. Infrafamilial classification and relationships have been controversial partially due to the lack of a robust phylogenetic framework. Including all genera except Indosinia and Perissocarpa and DNA sequence data for five DNA regions (ITS, matK, ndhF, rbcL, trnL-F), we provide for the first time a nearly complete molecular phylogenetic analysis of Ochnaceae s.l. resolving most of the phylogenetic backbone of the family. Based on this, we present a new classification of Ochnaceae s.l., with Medusagynoideae and Quiinoideae included as subfamilies and the former subfamilies Ochnoideae and Sauvagesioideae recognized at the rank of tribe. Our data support a monophyletic Ochneae, but Sauvagesieae in the traditional circumscription is paraphyletic because Testulea emerges as sister to the rest of Ochnoideae, and the next clade shows Luxemburgia+Philacra as sister group to the remaining Ochnoideae. To avoid paraphyly, we classify Luxemburgieae and Testuleeae as new tribes. The African genus Lophira, which has switched between subfamilies (here tribes) in past classifications, emerges as sister to all other Ochneae. Thus, endosperm-free seeds and ovules with partly to completely united integuments (resulting in an apparently single integument) are characters that unite all members of that tribe. The relationships within its largest clade, Ochnineae (former Ochneae), are poorly resolved, but former Ochninae (Brackenridgea, Ochna) are polyphyletic. Within Sauvagesieae, the genus Sauvagesia in its broad circumscription is polyphyletic as Sauvagesia serrata is sister to a clade of Adenarake, Sauvagesia spp., and three other genera. Within Quiinoideae, in contrast to former phylogenetic hypotheses, Lacunaria and Touroulia form a clade that is sister to Quiina. Bayesian ancestral state reconstructions showed that zygomorphic flowers with adaptations to buzz-pollination (poricidal anthers), a syncarpous gynoecium (a near-apocarpous gynoecium evolved independently in Quiinoideae and Ochninae), numerous ovules, septicidal capsules, and winged seeds with endosperm are the ancestral condition in Ochnoideae. Although in some lineages poricidal anthers were lost secondarily, the evolution of poricidal superstructures secured the maintenance of buzz-pollination in some of these genera, indicating a strong selective pressure on keeping that specialized pollination system.

Glochidionyangchunense (Phyllanthaceae), a new species with discoid flowers from Guangdong Province, China.

Phyllanthodendron can be readily morphologically distinguished from Glochidion, but recent molecular evidence showed that Phyllanthodendron is paraphyletic due to Glochidion being nested within it. In this study, a new species of the former Phyllanthodendron is described and illustrated as Glochidionyangchunense Z.Q. Song & Gang Yao from the limestone areas of South China. This is a peculiar new species and morphologically distinguished by its discoid flowers, T-shaped disc segments, and glabrous flowering branches. A key to Glochidionyangchunense and related species in China is provided here.

The genome sequence of black poplar, Populus nigra subsp. betulifolia L., 1753 (Salicaceae).

We present a genome assembly from an individual Populus nigra subsp. betulifola (black poplar; Tracheophyta; Malpighiales; Salicaceae). The genome sequence is 413.2 megabases in span. Most of the assembly (99.73%) is scaffolded into 19 chromosomal pseudomolecules. Mitochondrial and plastid genomes were also assembled. Three mitochondrial assemblies have lengths of 281.85, 335.57 and 186.15 kilobases, and the plastid genome has a length of 156.37 kilobases.

Seven new species of Rinorea (Violaceae) from the Neotropics.

Over the course of revising the genus Rinorea (Violaceae) from Colombia, field observations and herbarium studies revealed seven new species. Several of the new species described here belong to species complexes that required examination of herbarium material from across the Neotropics. Each of the new species described here have oppositely arranged leaves and belong to Rinoreasect.Pubiflorae, a section restricted to the Neotropics. Two new species are segregated from the R.ovalifolia species complex: Rinoreachiribiquetensis from Chiribiquete National Park in the Colombian Amazon and Rinoreastevensii from the Orinoco River near the border of Colombia and Venezuela. Two new species are segregated from the Rinoreahirsuta species complex: Rinoreagaleanoae-bernalii and Rinoreacogolloi, both from the eastern slopes of the Andean Central Cordillera along the mid-Magdalena River Valley in Colombia. From the widely distributed R.pubiflora species complex, we segregated one new species, Rinoreacallejasii, from southeast Panama and the Chocó in Colombia. In addition to these five new taxa segregated from widely distributed species complexes, we discovered two previously unknown species with affinities to other Neotropical Rinorea. Rinoreaaymardii is described from the Alto Orinoco-Casiquiare Biosphere Reserve in Venezuela and most closely resembles R.melanodonta from Colombia. Rinoreabetancurii is segregated from R.macrocarpa and occurs in the Amazonian Regions of Colombia, Brazil, Peru and Venezuela. In this study, we provide descriptions, illustrations and distribution maps of the new species and make preliminary assessments of the risk of extinction using the IUCN Red List Categories and Criteria. We also furnish an identification key to the species of Rinoreasect.Pubiflorae in Colombia.

The genome sequence of the Annual Mercury, Mercurialis annua L., 1753 (Euphorbiaceae).

We present a genome assembly from a diploid female Mercurialis annua (the Annual Mercury; Tracheophyta; Magnoliopsida; Malpighiales; Euphorbiaceae). The genome sequence is 453.2 megabases in span. Most of the assembly is scaffolded into 8 chromosomal pseudomolecules, including the X chromosome. The organelle genomes have also been assembled, and the mitochondrial genome is 435.28 kilobases in length, while the plastid genome is 169.65 kilobases in length.

A taxonomic revision of GarciniasectionGarcinia (Clusiaceae) in Thailand.

GarciniasectionGarcinia (Clusiaceae) is revised for Thailand with three species and one variety, i.e., two native species: G.celebica and G.exigua, and one cultivated species: G.mangostanavar.mangostana. Detailed morphological descriptions, illustrations, and an identification key to the species are presented, along with notes on distributions, habitats and ecology, phenology, conservation assessments, etymology, vernacular names, uses, and specimens examined. The section is recognized by its terminal inflorescences of simple cymes, or sometimes a solitary flower; flowers with 4 sepals and 4 petals; male flowers often with a pistillode, and stamens united into a single 4-lobed or 4-angled bundle, and with 2-thecous anthers; usually multilocular ovaries and stigmas with distinct or weak lobes and smooth or rough; and fruits with a smooth surface. Three associated synonyms of G.celebica: G.ferrea, G.basacensis, and G.hombroniana, are lectotypified here in a second-step. In Thailand, Garciniacelebica is found in a very wide variety of habitats, at elevations of 0-1,500 m amsl., and is known to be naturally distributed in all floristic regions. G.exigua is found in dry evergreen forest on limestone hills and in littoral dry evergreen forest on limestone hills, at elevations of 50-100 m amsl. in Krabi Province, the peninsular region. G.mangostanavar.mangostana is found only in cultivation. Garciniaexigua has a conservation status of Vulnerable [VU B2ab(iii)] and the other two species have a conservation status of Least Concern [LC]. The fleshy pulp surrounding the seeds of two species, G.celebica and G.mangostanavar.mangostana is edible and has a sweet-sour taste.

A taxonomic revision of Rhizophora L. (Rhizophoraceae) in Thailand.

A taxonomic revision of Rhizophora L. (Rhizophoraceae) in Thailand is presented. Two species, R. apiculata Blume and R. mucronata Poir., are enumerated with updated morphological descriptions, illustrations and a taxonomic identification key, together with notes on distributions, habitats and ecology, phenology, conservation assessments, etymology, vernacular names, uses, and specimens examined. Three names in Rhizophora, are lectotypified: R. apiculata and two associated synonyms of R. mucronata (i.e., R. latifolia Miq. and R. macrorrhiza Griff.). R. longissima Blanco, a synonym of R. mucronata, is neotypified. All two Rhizophora species have a conservation assessment of Least Concern (LC). Based on the morphological identification, these two species can be distinguished from one another by the shape and width of the leaf laminae and the length of a terminal stiff point of the leaf laminae; the type and position of the inflorescences and the number of flowers per inflorescence; the character and color of the bracteoles; the presence or absence of the flower pedicels; the shape of the mature flower buds; the shape, color, and texture of the sepals; the shape, character, and the presence or absence of hairs of the petals; the number of stamens per flower; the size of the fruits; the color and size of the hypocotyls; the color and diameter of the cotyledonous cylindrical tubes; and the color of the colleters and exudate. The thick cuticles, sunken stomata, large hypodermal cells, and cork warts are adaptive anatomical features of leaves in Rhizophora that live in the mangrove environments. The pollen grains of Thai Rhizophora species are tricolporate, prolate spheroidal or oblate spheroidal shapes, small-sized, and reticulate exine sculpturing.

The complete mitochondrial genome of the biodiesel plant Jatropha curcas L.

Jatropha curcas (Linnaeus, 1753) is a plant species in the order Malpighiales and the family Euphorbiaceae and is native to the tropical regions of America, such as Mexico and Argentina. Currently, this plant species inhabits tropical and subtropical regions of the world. Jatropha has been widely used as a biofuel plant to produce high-quality diesel engine fuel. In this study, the complete mitochondrial genome sequence of J. curcas was assembled into 561,839 bp circular nucleotides with a GC content of 44.6%. The mitochondrial genome of J. curcas comprises 33 known protein-coding genes, 22 tRNA genes, three rRNA genes, one ncRNA gene, and 85 open reading frame genes. Phylogenetic analysis showed this species is closely related to the castor bean (Ricinus communis).

Seed morphology of Hypericum (Hypericaceae) in China and its taxonomic significance.

The seed morphology of 40 taxa within the genus Hypericum (Hypericaceae) from China, representing 9 sections of the genus, was examined using both Light and Scanning Electron Microscopy to evaluate the taxonomic relevance of macro- and micro-morphological features. Details articulating variation in seed size, color, shape, appendages, and seed coat ornamentation are described, illustrated, and compared, and their taxonomic importance is discussed. Seeds were generally brown in color and cylindric-ellipsoid to prolonged cylindric in shape. Seed size displayed wide variation, ranging from 0.37-1.91 mm in length and 0.12-0.75 mm in width. Seed appendages were observed as a characteristic morphological feature. Seed surface ornamentation has high phenotypic plasticity, and four types (reticulate, foveolate, papillose, and ribbed) can be recognized. In general, seed color and shape have limited taxonomic significance. However, some other features represent informative characters that can be used efficiently in distinguishing the studied taxa at the section and/or species levels. The findings illustrate that considerable taxonomic knowledge can be obtained by investigating the seed features of Hypericum, and the use of Scanning Electron Microscopy can reveal inconspicuous morphological affinities among species and play a role in taxonomic and systematic studies of the genus Hypericum. RESEARCH HIGHLIGHTS: Macro- and micro-morphological features of seeds of 40 Hypericum taxa from China were examined using Light and Scanning Electron Microscopy, providing the first broad study regarding seed morphology for Hypericum from China. Details and variations of seed size, shape, color, surface ornamentation, and appendages are fully presented. Seed features and their variation have important taxonomic significance at the section and/or species levels within Hypericum.

A taxonomic revision of Garcinia section Xanthochymus (Clusiaceae) in Thailand.

Garcinia section Xanthochymus (Clusiaceae) is revised for Thailand with four native species, i.e., G. dulcis, G. nervosa, G. prainiana, and G. xanthochymus. All species are described with updated morphological descriptions, illustrations, and an identification key, together with notes on distributions, distribution maps, habitats and ecology, phenology, conservation assessments, etymology, vernacular names, uses, and specimens examined. Four taxa, G. andamanica, G. andamanica var. pubescens, G. cambodgiensis and G. vilersiana, are synonymized under G. dulcis, and two taxa, G. nervosa var. pubescens and G. spectabilis, are newly synonymized under G. nervosa. Nine names are lectotypified: G. dulcis and its associated synonyms (G. cambodgiensis and G. vilersiana), G. nervosa and its associated synonyms (G. andersonii, G. nervosa var. pubescens, and G. spectabilis), G. prainiana, and G. xanthochymus. All species have a conservation assessment of Least Concern (LC). The fruits of all species are edible and have a sour or sweet-sour taste.

Deciphering the Heteropterys pannosa species complex (Malpighiaceae).

We describe three new species of Malpighiaceae that are endemic to central Brazil and related to the Heteropterys pannosa complex, a group of xylopodiferous, unbranched subshrubs with fruit in mericarps that have a strongly reduced or no dorsal wing. Heteropterys tocantinensis is more common in eastern Tocantins State and on the border with Bahia State, and there are a few records from Mato Grosso State. Heteropterys veadeirensis is restricted to northern Goiás State and H. walteri has a wider distribution, occurring in some municipalities in northern Goiás and southern Tocantins. Additionally, we also provide detailed redescriptions of H. pannosa and H. rosmarinifolia, the two previously known species in this complex. All species are considered Endangered (EN) based on IUCN criteria, especially due to the low area of occupancy. Illustrations, distribution maps, and information about phenology and habitat are also provided for all taxa.

Untargeted Metabolomics Sheds Light on the Diversity of Major Classes of Secondary Metabolites in the Malpighiaceae Botanical Family.

Natural products produced by plants are one of the most investigated natural sources, which substantially contributed to the development of the natural products field. Even though these compounds are widely explored, the literature still lacks comprehensive investigations aiming to explore the evolution of secondary metabolites produced by plants, especially if classical methodologies are employed. The development of sensitive hyphenated techniques and computational tools for data processing has enabled the study of large datasets, being valuable assets for chemosystematic studies. Here, we describe a strategy for chemotaxonomic investigations using the Malpighiaceae botanical family as a model. Our workflow was based on MS/MS untargeted metabolomics, spectral searches, and recently described in silico classification tools, which were mapped into the latest molecular phylogeny accepted for this family. The metabolomic analysis revealed that different ionization modes and extraction protocols significantly impacted the chemical profiles, influencing the chemotaxonomic results. Spectral searches within public databases revealed several clades or genera-specific molecular families, being potential chemical markers for these taxa, while the in silico classification tools were able to expand the Malpighiaceae chemical space. The classes putatively annotated were used for ancestral character reconstructions, which recovered several classes of metabolites as homoplasies (i.e., non-exclusive) or synapomorphies (i.e., exclusive) for all sampled clades and genera. Our workflow combines several approaches to perform a comprehensive evolutionary chemical study. We expect it to be used on further chemotaxonomic investigations to expand chemical knowledge and reveal biological insights for compounds classes in different biological groups.

Clarifying the nomenclatural history of Tovomitopsis, a Brazilian endemic genus of Clusiaceae.

Tovomitopsis Planch. & Triana is a Brazilian Atlantic Forest endemic genus composed of two species: T.paniculata (Spreng.) Planch. & Triana and T.saldanhae Engl. An investigation was conducted to clarify the nomenclatural history of Tovomitopsis. We report the results of this investigation, provide an updated description of the genus, and propose lectotypes for T.paniculata and its synonyms: Tovomitafoliosa C.Presl and Tovomitapaniculata Cambess. We also propose lectotypes for T.saldanhae and for the new synonym Clusiaangustifolia Engl.

Heteropterys rosmarinifolia, a new species of Malpighiaceae with verticillate leaves from savannas grasslands of central Brazil.

A new species of Heteropterys belonging to the Parabanisteria informal group is described for the savannas grasslands of the Serra do Tombador Natural Reserve, municipality of Cavalcante, State of Goiás, Brazil. Heteropterys rosmarinifolia is easily distinguished from the remaining Brazilian species of the Parabanisteria informal group due to its stems unbranched, leaves associated with the inflorescence verticillate, blades linear to very narrowly oblong, strongly conduplicate, ascending to upright, and sparsely sericeous to glabrous at age. We present a complete morphological description for the new species, photographic plates, comments on its distribution, ecology, and taxonomy, besides an identification key to the species of Heteropterys from the Parabanisteria group from the State of Goiás, Brazil.

Characterization and Dynamics of Intracellular Gene Transfer in Plastid Genomes of Viola (Violaceae) and Order Malpighiales.

Functional gene transfer from organelles to the nucleus, known as intracellular gene transfer (IGT), is an ongoing process in flowering plants. The complete plastid genomes (plastomes) of two Ulleung island endemic violets, Viola ulleungdoensis and V. woosanensis, were characterized, revealing a lack of the plastid-encoded infA, rpl32, and rps16 genes. In addition, functional replacement of the three plastid-encoded genes in the nucleus was confirmed within the genus Viola and the order Malpighiales. Three strategies for the acquisition of a novel transit peptide for successful IGT were identified in the genus Viola. Nuclear INFA acquired a novel transit peptide with very low identity between these proteins, whereas the nuclear RPL32 gene acquired an existing transit peptide via fusion with the nuclear-encoded plastid-targeted SOD gene (Cu-Zn superoxide dismutase superfamily) as one exon, and translated both proteins in the cytosol using alternative mRNA splicing. Nuclear RPS16 contains an internal transit peptide without an N-terminal extension. Gene loss or pseudogenization of the plastid-borne rpl32 and rps16 loci was inferred to occur in the common ancestor of the genus Viola based on an infrageneric phylogenetic framework in Korea. Although infA was lost in the common ancestor of the order Malpighiales, the rpl32 and rps16 genes were lost multiple times independently within the order. Our current study sheds additional light on plastid genome composition and IGT mechanisms in the violet genus and in the order Malpighiales.

Resolving Recalcitrant Clades in the Pantropical Ochnaceae: Insights From Comparative Phylogenomics of Plastome and Nuclear Genomic Data Derived From Targeted Sequencing.

Plastid DNA sequence data have been traditionally widely used in plant phylogenetics because of the high copy number of plastids, their uniparental inheritance, and the blend of coding and non-coding regions with divergent substitution rates that allow the reconstruction of phylogenetic relationships at different taxonomic ranks. In the present study, we evaluate the utility of the plastome for the reconstruction of phylogenetic relationships in the pantropical plant family Ochnaceae (Malpighiales). We used the off-target sequence read fraction of a targeted sequencing study (targeting nuclear loci only) to recover more than 100 kb of the plastid genome from the majority of the more than 200 species of Ochnaceae and all but two genera using de novo and reference-based assembly strategies. Most of the recalcitrant nodes in the family's backbone were resolved by our plastome-based phylogenetic inference, corroborating the most recent classification system of Ochnaceae and findings from a phylogenomic study based on nuclear loci. Nonetheless, the phylogenetic relationships within the major clades of tribe Ochnineae, which comprise about two thirds of the family's species diversity, received mostly low support. Generally, the phylogenetic resolution was lowest at the infrageneric level. Overall there was little phylogenetic conflict compared to a recent analysis of nuclear loci. Effects of taxon sampling were invoked as the most likely reason for some of the few well-supported discords. Our study demonstrates the utility of the off-target fraction of a target enrichment study for assembling near-complete plastid genomes for a large proportion of samples.