Re-evaluating monotypic Eleutherostylis from New Guinea and the Moluccas and its inclusion in Grewia (Malvaceae, Grewioideae).

Morphological and molecular phylogenetic evidence indicate that Eleutherostylis Burret (Malvaceae, Grewioideae), a monotypic genus described from New Guinea, is best considered a synonym of Grewia L., a species-rich genus widespread across the Paleotropics and found in Africa, Arabia, Asia, Australia and the western Pacific. A new combination, based on E.renistipulata Burret, G.renistipulata (Burret) Dorr, comb. nov., is proposed. Original material of the basionym could not be located and a neotype is designated. A lectotype is designated for G.morotaiensis Kosterm., a synonym of G.renistipulata.

A new, disjunct species of Bahiana (Euphorbiaceae, Acalyphoideae): Phytogeographic connections between the seasonally dry tropical forests of Peru and Brazil, and a review of spinescence in the family.

Bahiana is expanded from 1 to 2 species with the description of B.occidentalis K. Wurdack, sp. nov. as a new endemic of the seasonally dry tropical forests (SDTFs) of Peru. The disjunct distribution of Bahiana with populations of B.occidentalis on opposite sides of the Andes in northwestern Peru (Tumbes, San Martín) and B.pyriformis in eastern Brazil (Bahia) adds to the phytogeographic links among the widely scattered New World SDTFs. Although B.occidentalis remains imperfectly known due to the lack of flowering collections, molecular phylogenetic results from four loci (plastid matK, rbcL, and trnL-F; and nuclear ITS) unite the two species as does gross vegetative morphology, notably their spinose stipules, and androecial structure. Spinescence in Euphorbiaceae was surveyed and found on vegetative organs in 25 genera, which mostly have modified sharp branch tips. Among New World taxa, spines that originate from stipule modifications only occur in Bahiana and Acidocroton, while the intrastipular spines of Philyra are of uncertain homologies.

Monadelpha (Euphorbiaceae, Plukenetieae), a new genus of Tragiinae from the Amazon rainforest of Venezuela and Brazil.

Monadelpha L.J.Gillespie & Card.-McTeag., gen. nov., is described as a new member of Euphorbiaceae tribe Plukenetieae subtribe Tragiinae, to accommodate Tragia guayanensis, a species known from western Amazonas, Venezuela and, newly reported here, from Amazonas, Brazil. The genus is unique in the subtribe for having 5-colpate pollen and staminate flowers with filaments entirely connate into an elongate, cylindrical staminal column terminated by a tight cluster of anthers. Phylogenetic analyses based on nuclear rDNA ITS and sampling 156 accessions across the diversity of Tragiinae (all 12 genera and 77 of ~195 species) also support Monadelpha as a distinct lineage that is separate from Tragia. A revised key to the genera of Tragiinae in South America and Central America is provided.

Pseudoflowers produced by Fusarium xyrophilum on yellow-eyed grass (Xyris spp.) in Guyana: A novel floral mimicry system?

Pseudoflower formation is arguably the rarest outcome of a plant-fungus interaction. Here we report on a novel putative floral mimicry system in which the pseudoflowers are composed entirely of fungal tissues in contrast to modified leaves documented in previous mimicry systems. Pseudoflowers on two perennial Xyris species (yellow-eyed grass, X. setigera and X. surinamensis) collected from savannas in Guyana were produced by Fusarium xyrophilum, a novel Fusarium species. These pseudoflowers mimic Xyris flowers in gross morphology and are ultraviolet reflective. Axenic cultures of F. xyrophilum produced two pigments that had fluorescence emission maxima in light ranges that trichromatic insects are sensitive to and volatiles known to attract insect pollinators. One of the volatiles emitted by F. xyrophilum cultures (i.e., 2-ethylhexanol) was also detected in the head space of X. laxifolia var. iridifolia flowers, a perennial species native to the New World. Results of microscopic and PCR analyses, combined with examination of gross morphology of the pseudoflowers, provide evidence that the fungus had established a systemic infection in both Xyris species, sterilized them and formed fungal pseudoflowers containing both mating type idiomorphs. Fusarium xyrophilum cultures also produced the auxin indole-3-acetic acid (IAA) and the cytokinin isopentenyl adenosine (iPR). Field observations revealed that pseudoflowers and Xyris flowers were both visited by bees. Together, the results suggest that F. xyrophilum pseudoflowers are a novel floral mimicry system that attracts insect pollinators, via visual and olfactory cues, into vectoring its conidia, which might facilitate outcrossing of this putatively heterothallic fungus and infection of previously uninfected plants.

Fusarium xyrophilum, sp. nov., a member of the Fusarium fujikuroi species complex recovered from pseudoflowers on yellow-eyed grass (Xyris spp.) from Guyana.

We report on the discovery and characterization of a novel Fusarium species that produced yellow-orange pseudoflowers on Xyris spp. (yellow-eyed grass; Xyridaceae) growing in the savannas of the Pakaraima Mountains of western Guyana. The petaloid fungal structures produced on infected plants mimic host flowers in gross morphology. Molecular phylogenetic analyses of full-length RPB1 (RNA polymerase largest subunit), RPB2 (RNA polymerase second largest subunit), and TEF1 (elongation factor 1-α) DNA sequences mined from genome sequences resolved the fungus, described herein as F. xyrophilum, sp. nov., as sister to F. pseudocircinatum within the African clade of the F. fujikuroi species complex. Results of a polymerase chain reaction (PCR) assay for mating type idiomorph revealed that single-conidial isolates of F. xyrophilum had only one of the MAT idiomorphs (MAT1-1 or MAT1-2), which suggests that the fungus may have a heterothallic sexual reproductive mode. BLASTn searches of whole-genome sequence of three strains of F. xyrophilum indicated that it has the genetic potential to produce secondary metabolites, including phytohormones, pigments, and mycotoxins. However, a polyketide-derived pigment, 8-O-methylbostrycoidin, was the only metabolite detected in cracked maize kernel cultures. When grown on carnation leaf agar, F. xyrophilum is phenotypically distinct from other described Fusarium species in that it produces aseptate microconidia on erect indeterminate synnemata that are up to 2 mm tall and it does not produce multiseptate macroconidia.

Systematics and relationships of Tryssophyton (Melastomataceae), with a second species from the Pakaraima Mountains of Guyana.

The systematics of Tryssophyton, herbs endemic to the Pakaraima Mountains of western Guyana, is reviewed and Tryssophyton quadrifolius K.Wurdack & Michelang., sp. nov. from the summit of Kamakusa Mountain is described as the second species in the genus. The new species is distinguished from its closest relative, Tryssophyton merumense, by striking vegetative differences, including number of leaves per stem and leaf architecture. A phylogenetic analysis of sequence data from three plastid loci and Melastomataceae-wide taxon sampling is presented. The two species of Tryssophyton are recovered as monophyletic and associated with mostly Old World tribe Sonerileae. Fruit, seed and leaf morphology are described for the first time, biogeography is discussed and both species are illustrated.

Insights on the systematics and morphology of Humiriaceae (Malpighiales): androecial and extrafloral nectary variation, two new combinations, and a new Sacoglottis from Guyana.

Humiriaceae have had little recent comparative morphological study except for their distinctive fruits. We surveyed the diversity of stamen structures in the family with consideration of dehiscence patterns and the evolutionary transitions between tetra- and disporangiate anthers. Novel interpretations of floral morphology support new combinations (Duckesialiesneri K.Wurdack & C.E.Zartman, comb. nov. and Vantaneaspiritu-sancti K.Wurdack & C.E.Zartman, comb. nov.) for two species formerly in Humiriastrum. We investigated all eleven species of Sacoglottis for diagnostic features that may contribute to better species delimitations, and describe Sacoglottisperryi K.Wurdack & C.E.Zartman, sp. nov. as an endemic of the Pakaraima Mountains in western Guyana. Finally, our survey across Humiriaceae for extrafloral nectaries (EFNs) revealed their presence on leaves of all extant species as adaxial basilaminar and/or abaxial embedded glands, in addition to the frequent occurrence of marginal glandular setae. The significance of inter-generic variation in gland position and anther morphology within the family are discussed.

Seed size evolution and biogeography of Plukenetia (Euphorbiaceae), a pantropical genus with traditionally cultivated oilseed species.

BACKGROUND: Plukenetia is a small pantropical genus of lianas and vines with variably sized edible oil-rich seeds that presents an ideal system to investigate neotropical and pantropical diversification patterns and seed size evolution. We assessed the biogeography and seed evolution of Plukenetia through phylogenetic analyses of a 5069 character molecular dataset comprising five nuclear and two plastid markers for 86 terminals in subtribe Plukenetiinae (representing 20 of ~ 23 Plukenetia species). Two nuclear genes, KEA1 and TEB, were used for phylogenetic reconstruction for the first time. Our goals were: (1) produce a robust, time-dependent evolutionary framework for Plukenetia using BEAST; (2) reconstruct its biogeographical history with ancestral range estimation in BIOGEOBEARS; (3) define seed size categories; (4) identify patterns of seed size evolution using ancestral state estimation; and (5) conduct regression analyses with putative drivers of seed size using the threshold model. RESULTS: Plukenetia was resolved into two major groups, which we refer to as the pinnately- and palmately-veined clades. Our analyses suggest Plukenetia originated in the Amazon or Atlantic Forest of Brazil during the Oligocene (28.7 Mya) and migrated/dispersed between those regions and Central America/Mexico throughout the Miocene. Trans-oceanic dispersals explain the pantropical distribution of Plukenetia, including from the Amazon to Africa in the Early Miocene (17.4 Mya), followed by Africa to Madagascar and Africa to Southeast Asia in the Late Miocene (9.4 Mya) and Pliocene (4.5 Mya), respectively. We infer a single origin of large seeds in the ancestor of Plukenetia. Seed size fits a Brownian motion model of trait evolution and is moderately to strongly associated with plant size, fruit type/dispersal syndrome, and seedling ecology. Biome shifts were not drivers of seed size, although there was a weak association with a transition to fire prone semi-arid savannas. CONCLUSIONS: The major relationships among the species of Plukenetia are now well-resolved. Our biogeographical analyses support growing evidence that many pantropical distributions developed by periodic trans-oceanic dispersals throughout the Miocene and Pliocene. Selection on a combination of traits contributed to seed size variation, while movement between forest edge/light gap and canopy niches likely contributed to the seed size extremes in Plukenetia.

A new disjunct species of Eriolaena (Malvaceae, Dombeyoideae) from Continental Africa.

Eriolaenarulkensii Dorr, sp. nov. is described and illustrated. This attractive shrub is endemic to coastal Mozambique. The new species has apically winged seeds, which place it in a group of Malvaceae (Dombeyoideae) that is found in Asia and Madagascar and which had not previously been found in continental Africa.

A new large-flowered species of Andeimalva (Malvaceae, Malvoideae) from Peru.

Andeimalvaperuviana Dorr & C.Romero, sp. nov., the third Peruvian endemic in a small genus of five species, is described and illustrated from a single collection made at high elevation on the eastern slopes of the Andes. Molecular phylogenetic analyses of nuclear ribosomal ITS sequence data resolve a group of northern species of Andeimalva found in Bolivia and Peru from the morphologically very different southern A.chilensis. The new species bears the largest flowers of any Andeimalva and is compared with Bolivian A.mandonii. A revised key to the genus is presented.

A new disjunct Dendrothrix (Euphorbiaceae, tribe Hippomaneae): a Guiana Shield element in sub-Andean cordilleras of Ecuador and Peru.

Dendrothrix condorensis K.Wurdack, sp. nov. from the sub-Andean cordilleras of Ecuador and Peru is described and illustrated. The new species is geographically widely separated from its likely closest relative, D. yutajensis, which is endemic to the Guiana Shield region of southern Venezuela and adjacent Brazil, and notably differs in leaf morphology. Vegetative (i.e., epidermal micropapillae, trichomes) and reproductive (i.e., cymule glands, flowers, pollen) micromorphological features were examined with SEM. Rare tristaminate flowers were documented among the typical bistaminate ones. Seeds and diagnostic features among the four species of Dendrothrix are compared.

Incadendron : a new genus of Euphorbiaceae tribe Hippomaneae from the sub-Andean cordilleras of Ecuador and Peru.

Incadendron esseri K.Wurdack & Farfan, gen. & sp. nov., from the wet sub-Andean cordilleras of Ecuador (Cordillera del Cóndor) and Peru (Cusco, Oxapampa) is described and illustrated. This recently discovered large canopy tree with a narrow elevational range presents an unusual combination of rare morphological characters in Hippomaneae including mucilage-secreting sheathing stipules, conduplicate ptyxis, and large, woody fruits. The broader significance of these characters in Hippomaneae is discussed. The morphology and anatomy of Incadendron were investigated, highlighting its fruit similarities with Guiana Shield endemic Senefelderopsis, and the systematics value of ptyxis variation, which remains poorly studied for the family.

A new tiny-leaved species of Raveniopsis (Rutaceae) from the Pakaraima Mountains of Guyana.

Raveniopsis microphyllus K.Wurdack, sp. nov., a new species known only from a single peak in the Pakaraima Mtns. of Guyana, is described and illustrated. This white-flowered shrub adds to the many narrow-endemic Guiana Shield species in the genus, and is unique in bearing small, trifoliate, sclerophyllous leaves. Leaf anatomy and surface micromorphology of the new species were examined to document its montane adaptations. The multiple trichome types of the leaves and flowers of new species were characterized, and the systematics value of the considerable foliar trichome variation in Raveniopsis is discussed.

Evolutionary bursts in Euphorbia (Euphorbiaceae) are linked with photosynthetic pathway.

The mid-Cenozoic decline of atmospheric CO2 levels that promoted global climate change was critical to shaping contemporary arid ecosystems. Within angiosperms, two CO2 -concentrating mechanisms (CCMs)-crassulacean acid metabolism (CAM) and C4 -evolved from the C3 photosynthetic pathway, enabling more efficient whole-plant function in such environments. Many angiosperm clades with CCMs are thought to have diversified rapidly due to Miocene aridification, but links between this climate change, CCM evolution, and increased net diversification rates (r) remain to be further understood. Euphorbia (∼2000 species) includes a diversity of CAM-using stem succulents, plus a single species-rich C4 subclade. We used ancestral state reconstructions with a dated molecular phylogeny to reveal that CCMs independently evolved 17-22 times in Euphorbia, principally from the Miocene onwards. Analyses assessing among-lineage variation in r identified eight Euphorbia subclades with significantly increased r, six of which have a close temporal relationship with a lineage-corresponding CCM origin. Our trait-dependent diversification analysis indicated that r of Euphorbia CCM lineages is approximately threefold greater than C3 lineages. Overall, these results suggest that CCM evolution in Euphorbia was likely an adaptive strategy that enabled the occupation of increased arid niche space accompanying Miocene expansion of arid ecosystems. These opportunities evidently facilitated recent, replicated bursts of diversification in Euphorbia.

Phylogenomics and a posteriori data partitioning resolve the Cretaceous angiosperm radiation Malpighiales.

The angiosperm order Malpighiales includes ~16,000 species and constitutes up to 40% of the understory tree diversity in tropical rain forests. Despite remarkable progress in angiosperm systematics during the last 20 y, relationships within Malpighiales remain poorly resolved, possibly owing to its rapid rise during the mid-Cretaceous. Using phylogenomic approaches, including analyses of 82 plastid genes from 58 species, we identified 12 additional clades in Malpighiales and substantially increased resolution along the backbone. This greatly improved phylogeny revealed a dynamic history of shifts in net diversification rates across Malpighiales, with bursts of diversification noted in the Barbados cherries (Malpighiaceae), cocas (Erythroxylaceae), and passion flowers (Passifloraceae). We found that commonly used a priori approaches for partitioning concatenated data in maximum likelihood analyses, by gene or by codon position, performed poorly relative to the use of partitions identified a posteriori using a Bayesian mixture model. We also found better branch support in trees inferred from a taxon-rich, data-sparse matrix, which deeply sampled only the phylogenetically critical placeholders, than in trees inferred from a taxon-sparse matrix with little missing data. Although this matrix has more missing data, our a posteriori partitioning strategy reduced the possibility of producing multiple distinct but equally optimal topologies and increased phylogenetic decisiveness, compared with the strategy of partitioning by gene. These approaches are likely to help improve phylogenetic resolution in other poorly resolved major clades of angiosperms and to be more broadly useful in studies across the Tree of Life.

Horizontal transfer of expressed genes in a parasitic flowering plant.

BACKGROUND: Recent studies have shown that plant genomes have potentially undergone rampant horizontal gene transfer (HGT). In plant parasitic systems HGT appears to be facilitated by the intimate physical association between the parasite and its host. HGT in these systems has been invoked when a DNA sequence obtained from a parasite is placed phylogenetically very near to its host rather than with its closest relatives. Studies of HGT in parasitic plants have relied largely on the fortuitous discovery of gene phylogenies that indicate HGT, and no broad systematic search for HGT has been undertaken in parasitic systems where it is most expected to occur. RESULTS: We analyzed the transcriptomes of the holoparasite Rafflesia cantleyi Solms-Laubach and its obligate host Tetrastigma rafflesiae Miq. using phylogenomic approaches. Our analyses show that several dozen actively transcribed genes, most of which appear to be encoded in the nuclear genome, are likely of host origin. We also find that hundreds of vertically inherited genes (VGT) in this parasitic plant exhibit codon usage properties that are more similar to its host than to its closest relatives. CONCLUSIONS: Our results establish for the first time a substantive number of HGTs in a plant host-parasite system. The elevated rate of unidirectional host-to- parasite gene transfer raises the possibility that HGTs may provide a fitness benefit to Rafflesia for maintaining these genes. Finally, a similar convergence in codon usage of VGTs has been shown in microbes with high HGT rates, which may help to explain the increase of HGTs in these parasitic plants.

Phylogenetics and the evolution of major structural characters in the giant genus Euphorbia L. (Euphorbiaceae).

Euphorbia is among the largest genera of angiosperms, with about 2000 species that are renowned for their remarkably diverse growth forms. To clarify phylogenetic relationships in the genus, we used maximum likelihood, bayesian, and parsimony analyses of DNA sequence data from 10 markers representing all three plant genomes, averaging more than 16kbp for each accession. Taxon sampling included 176 representatives from Euphorbioideae (including 161 of Euphorbia). Analyses of these data robustly resolve a backbone topology of four major, subgeneric clades--Esula, Rhizanthium, Euphorbia, and Chamaesyce--that are successively sister lineages. Ancestral state reconstructions of six reproductive and growth form characters indicate that the earliest Euphorbia species were likely woody, non-succulent plants with helically arranged leaves and 5-glanded cyathia in terminal inflorescences. The highly modified growth forms and reproductive features in Euphorbia have independent origins within the subgeneric clades. Examples of extreme parallelism in trait evolution include at least 14 origins of xeromorphic growth forms and at least 13 origins of seed caruncles. The evolution of growth form and inflorescence position are significantly correlated, and a pathway of evolutionary transitions is supported that has implications for the evolution of Euphorbia xerophytes of large stature. Such xerophytes total more than 400 species and are dominants of vegetation types throughout much of arid Africa and Madagascar.

Angiosperm phylogeny: 17 genes, 640 taxa.

PREMISE OF THE STUDY: Recent analyses employing up to five genes have provided numerous insights into angiosperm phylogeny, but many relationships have remained unresolved or poorly supported. In the hope of improving our understanding of angiosperm phylogeny, we expanded sampling of taxa and genes beyond previous analyses. METHODS: We conducted two primary analyses based on 640 species representing 330 families. The first included 25260 aligned base pairs (bp) from 17 genes (representing all three plant genomes, i.e., nucleus, plastid, and mitochondrion). The second included 19846 aligned bp from 13 genes (representing only the nucleus and plastid). KEY RESULTS: Many important questions of deep-level relationships in the nonmonocot angiosperms have now been resolved with strong support. Amborellaceae, Nymphaeales, and Austrobaileyales are successive sisters to the remaining angiosperms (Mesangiospermae), which are resolved into Chloranthales + Magnoliidae as sister to Monocotyledoneae + [Ceratophyllaceae + Eudicotyledoneae]. Eudicotyledoneae contains a basal grade subtending Gunneridae. Within Gunneridae, Gunnerales are sister to the remainder (Pentapetalae), which comprises (1) Superrosidae, consisting of Rosidae (including Vitaceae) and Saxifragales; and (2) Superasteridae, comprising Berberidopsidales, Santalales, Caryophyllales, Asteridae, and, based on this study, Dilleniaceae (although other recent analyses disagree with this placement). Within the major subclades of Pentapetalae, most deep-level relationships are resolved with strong support. CONCLUSIONS: Our analyses confirm that with large amounts of sequence data, most deep-level relationships within the angiosperms can be resolved. We anticipate that this well-resolved angiosperm tree will be of broad utility for many areas of biology, including physiology, ecology, paleobiology, and genomics.

Malpighiales phylogenetics: Gaining ground on one of the most recalcitrant clades in the angiosperm tree of life.

The eudicot order Malpighiales contains ∼16000 species and is the most poorly resolved large rosid clade. To clarify phylogenetic relationships in the order, we used maximum likelihood, Bayesian, and parsimony analyses of DNA sequence data from 13 gene regions, totaling 15604 bp, and representing all three genomic compartments (i.e., plastid: atpB, matK, ndhF, and rbcL; mitochondrial: ccmB, cob, matR, nad1B-C, nad6, and rps3; and nuclear: 18S rDNA, PHYC, and newly developed low-copy EMB2765). Our sampling of 190 taxa includes representatives from all families of Malpighiales. These data provide greatly increased support for the recent additions of Aneulophus, Bhesa, Centroplacus, Ploiarium, and Rafflesiaceae to Malpighiales; sister relations of Phyllanthaceae + Picrodendraceae, monophyly of Hypericaceae, and polyphyly of Clusiaceae. Oxalidales + Huaceae, followed by Celastrales are successive sisters to Malpighiales. Parasitic Rafflesiaceae, which produce the world's largest flowers, are confirmed as embedded within a paraphyletic Euphorbiaceae. Novel findings show a well-supported placement of Ctenolophonaceae with Erythroxylaceae + Rhizophoraceae, sister-group relationships of Bhesa + Centroplacus, and the exclusion of Medusandra from Malpighiales. New taxonomic circumscriptions include the addition of Bhesa to Centroplacaceae, Medusandra to Peridiscaceae (Saxifragales), Calophyllaceae applied to Clusiaceae subfamily Kielmeyeroideae, Peraceae applied to Euphorbiaceae subfamily Peroideae, and Huaceae included in Oxalidales.

The complete nucleotide sequence of the cassava (Manihot esculenta) chloroplast genome and the evolution of atpF in Malpighiales: RNA editing and multiple losses of a group II intron.

The complete sequence of the chloroplast genome of cassava (Manihot esculenta, Euphorbiaceae) has been determined. The genome is 161,453 bp in length and includes a pair of inverted repeats (IR) of 26,954 bp. The genome includes 128 genes; 96 are single copy and 16 are duplicated in the IR. There are four rRNA genes and 30 distinct tRNAs, seven of which are duplicated in the IR. The infA gene is absent; expansion of IRb has duplicated 62 amino acids at the 3' end of rps19 and a number of coding regions have large insertions or deletions, including insertions within the 23S rRNA gene. There are 17 intron-containing genes in cassava, 15 of which have a single intron while two (clpP, ycf3) have two introns. The usually conserved atpF group II intron is absent and this is the first report of its loss from land plant chloroplast genomes. The phylogenetic distribution of the atpF intron loss was determined by a PCR survey of 251 taxa representing 34 families of Malpighiales and 16 taxa from closely related rosids. The atpF intron is not only missing in cassava but also from closely related Euphorbiaceae and other Malpighiales, suggesting that there have been at least seven independent losses. In cassava and all other sequenced Malphigiales, atpF gene sequences showed a strong association between C-to-T substitutions at nucleotide position 92 and the loss of the intron, suggesting that recombination between an edited mRNA and the atpF gene may be a possible mechanism for the intron loss.