Crown Node Research Articles

Genome-Wide Gene Birth-Death Dynamics Are Associated with Diet Breadth Variation in Lepidoptera.

Comparative analyses of gene birth-death dynamics have the potential to reveal gene families that played an important role in the evolution of morphological, behavioral, or physiological variation. Here, we used whole genomes of 30 species of butterflies and moths to identify gene birth-death dynamics among the Lepidoptera that are associated with specialist or generalist feeding strategies. Our work advances this field using a uniform set of annotated proteins for all genomes, investigating associations while correcting for phylogeny, and assessing all gene families rather than a priori subsets. We discovered that the sizes of several important gene families (e.g. those associated with pesticide resistance, xenobiotic detoxification, and/or protein digestion) are significantly correlated with diet breadth. We also found 22 gene families showing significant shifts in gene birth-death dynamics at the butterfly (Papilionoidea) crown node, the most notable of which was a family of pheromone receptors that underwent a contraction potentially linked with a shift to visual-based mate recognition. Our findings highlight the importance of uniform annotations, phylogenetic corrections, and unbiased gene family analyses in generating a list of candidate genes that warrant further exploration.

Molecular phylogeny and spatio‐temporal diversification of the Millettioid/Phaseoloid clade (Fabaceae: Papilionoideae)

AbstractThe Millettioid/Phaseoloid (or the Millettioid) clade is a major lineage of the subfamily Papilionoideae (Fabaceae) that is poorly understood in terms of its diversification and biogeographic history. To fill this gap, we generated a time‐calibrated phylogeny for 749 species representing c. 80% of the genera of this clade using nrDNA ITS, plastid matK, and plastome sequence (including 38 newly sequenced plastomes). Using this phylogenetic framework, we explored the clade's temporal diversification and reconstructed its ancestral areas and dispersal events. Our phylogenetic analyses support the monophyly of the Millettioid/Phaseoloid clade and four of its tribal lineages (Abreae, Desmodieae, Indigofereae, and Psoraleeae), while two tribal lineages sensu lato millettioids and phaseoloids are polyphyletic. The fossil‐calibrated dating analysis showed a nearly simultaneous divergence between the stem node (c. 62 Ma) and the crown node (c. 61 Ma) of the Millettioid/Phaseoloid clade in the Paleocene. The biogeographic analysis suggested that the clade originated in Africa and dispersed to Asia, Europe, Australia, and the Americas at different periods in the Cenozoic. We found evidence for shifts in diversification rates across the phylogeny of the Millettioid/Phaseoloid clade throughout the Cenozoic, with a rapid increase in net diversification rates since c. 10 Ma. Possible explanations for the present‐day species richness and distribution of the Millettioid/Phaseoloid clade include boreotropical migration, frequent intra‐ and intercontinental long‐distance dispersals throughout the Cenozoic, and elevated speciation rates following the Mid‐Miocene Climatic Optimum. Together, these results provide novel insights into major diversification patterns of the Millettioid/Phaseoloid clade, setting the stage for future evolutionary research on this important legume clade.

Diversification and dispersal in the Americas revealed by new phylogenies of the wrens and allies (Passeriformes: Certhioidea)

Abstract The passerine superfamily Certhioidea lacks a complete phylogeny despite decades of recognition as a clade and extensive systematic work within all its constituent families. Here, we inferred a near-complete species-level phylogeny of Certhioidea from a molecular supermatrix, including the first comprehensive sampling of the wrens (Troglodytidae), and used this phylogeny to infer its biogeographic and diversification histories. We also inferred an expanded phylogeny including nearly 100 putative phylospecies previously documented in the literature, and we found that including this diversity had notable impacts on the inferred evolutionary history of Certhioidea. This phylospecies-level tree documented a few instances of species paraphyly, some previously described in the literature and some novel. We found that Certhioidea originated largely in Eurasia and dispersed into North America five times in the last 20 million years, including at the origin of the “New World certhioids,” wrens and gnatcatchers, a clade herein named Orthourae. After this initial dispersal event, both wrens and gnatcatchers diversified extensively across the hemisphere, with both lineages repeatedly crossing between continents. However, we detected no notable impact of the formation of the Isthmus of Panama on the frequency of dispersal events between North and South America. The inclusion of phylospecies altered this biogeographic inference in some portions of the tree but overall was largely consistent. With species-level sampling, we found that diversification rates within Certhioidea were largely constant through time with a detectable deceleration toward the present. By contrast, phylospecies-level sampling recovered a different diversification history with a significant rate increase at the crown node of Orthourae after dispersing into the Americas and increased speciation rates particularly within the genera Polioptila and Henicorhina. This largely resolved phylogeny for Certhioidea has yielded important insights into the evolutionary history of this group and provides a framework for future comparative work on this fascinating clade.

Comparative Chloroplast Genomics of 21 Species in Zingiberales with Implications for Their Phylogenetic Relationships and Molecular Dating.

Zingiberales includes eight families and more than 2600 species, with many species having important economic and ecological value. However, the backbone phylogenetic relationships of Zingiberales still remain controversial, as demonstrated in previous studies, and molecular dating based on chloroplast genomes has not been comprehensively studied for the whole order. Herein, 22 complete chloroplast genomes from 21 species in Zingiberales were sequenced, assembled, and analyzed. These 22 genomes displayed typical quadripartite structures, which ranged from 161,303 bp to 163,979 bp in length and contained 111-112 different genes. The genome structures, gene contents, simple sequence repeats, long repeats, and codon usage were highly conserved, with slight differences among these genomes. Further comparative analysis of the 111 complete chloroplast genomes of Zingiberales, including 22 newly sequenced ones and the remaining ones from the national center for biotechnology information (NCBI) database, identified three highly divergent regions comprising ccsA, psaC, and psaC-ndhE. Maximum likelihood and Bayesian inference phylogenetic analyses based on chloroplast genome sequences found identical topological structures and identified a strongly supported backbone of phylogenetic relationships. Cannaceae was sister to Marantaceae, forming a clade that was collectively sister to the clade of (Costaceae, Zingiberaceae) with strong support (bootstrap (BS) = 100%, and posterior probability (PP) = 0.99-1.0); Heliconiaceae was sister to the clade of (Lowiaceae, Strelitziaceae), then collectively sister to Musaceae with strong support (BS = 94-100%, and PP = 0.93-1.0); the clade of ((Cannaceae, Marantaceae), (Costaceae, Zingiberaceae)) was sister to the clade of (Musaceae, (Heliconiaceae, (Lowiaceae, Strelitziaceae))) with robust support (BS = 100%, and PP = 1.0). The results of divergence time estimation of Zingiberales indicated that the crown node of Zingiberales occurred approximately 85.0 Mya (95% highest posterior density (HPD) = 81.6-89.3 million years ago (Mya)), with major family-level lineages becoming from 46.8 to 80.5 Mya. These findings proved that chloroplast genomes could contribute to the study of phylogenetic relationships and molecular dating in Zingiberales, as well as provide potential molecular markers for further taxonomic and phylogenetic studies of Zingiberales.

Biogeography of the Sunda Shelf revisited: Insights from Macaranga section Pruinosae (Euphorbiaceae)

The Southeast Asian region of Sundaland is among the world’s major biodiversity hotspots. The region’s biodiversity coupled with its complex and dynamic geographic and climatic histories makes it an ideal region to study the various factors that determine the diversification and distribution patterns of tropical biota. Here we investigate the biogeographic patterns in the partly myrmecophyticMacarangasectionPruinosaeto reveal some of the factors that play a role in determining the distribution of biota in Sundaland. We used single nucleotide polymorphisms (SNP) data derived from GBS, a next generation sequencing technique, in maximum likelihood and cluster analyses to determine phylogenetic relationships and population structures within this taxonomic section. Bayesian inference based on secondary calibration points and ancestral area reconstruction analyses were performed to infer spatial–temporal origins of the major lineages in the section. The results from these analyses were further substantiated using nuclear microsatellite data obtained from a broader sample set of two widely distributed species within the section:Macaranga giganteaandMacaranga pruinosa. Phylogenetic and cluster analyses reveal four well-defined, discrete species groups within sectionPruinosae, all of which but one originated in Borneo with the crown node dated at 3.58 mya. Biogeographic patterns within the species reveal a biogeographic barrier between east and west Sundaland besides bringing to light the role played by various geological factors, especially the Crocker Range, on Borneo. Patterns also reveal a biogeographic barrier between the Bangka/Belitung islands and Sumatra for ant-free, swamp-adapted species. This study provides evidence that geographic barriers, edaphic conditions, and ecological adaptations are tightly linked and that their mutual interaction determines the diversification and distribution of species.

Molecular and morphological evidence support the recognition of three genera within Radulaceae (Porellales: Marchantiophyta)

The discovery of new fossil bryophytes allows refined estimates of divergence times when the fossils have unambiguous synapomorphies enabling their confident assignment to nodes within molecular phylogenies. We use two fossil Radula species from Cretaceous age Burmese amber to estimate divergence times for Radula. One of these fossils, R. cretacea which has synapomorphies of subg. Odontoradula, has been used previously; the other, with relative synapomorphies of subg. Amentuloradula, has not. In combination these two fossils, when used to constrain the crown node age of their respective subgenera, result in median age estimates for the Radula crown node of 263 million years, under our preferred time-calibration scenario where subgeneric crown-node fossil assignments are coupled with a secondary maximum age constraint on the Porellales crown node. We explore other time-calibration scenarios, including deeper fossil assignments, and conclude that, under all, Radula as currently circumscribed is an outlier among land plant genera on the basis of its age. While this violates no established norms regarding how old genera should be, the absolute age, the relative ages of other families within Porellales, and the morphological distinctiveness of the two serial sister lineages, subg. Cladoradula, with seven species, and subg. Dactyloradula with one species, motivates our proposal to elevate these two subgenera to generic rank, resulting in three genera within the family Radulaceae. We provide diagnoses for genera, new combinations for species, and an emended circumscription for Radula, that reflect this change.

Out of Madagascar, repeatedly: The phylogenetics and biogeography of Dombeyoideae (Malvaceae s.l.)

Dispersals have been shown to be critical to the evolution of the long isolated but megadiverse flora of Madagascar and the surrounding islands of the western Indian Ocean, but we are just beginning to understand the directionality of these dispersals. With more than half of its species occurring in the western Indian Ocean region (WIOR), the paleotropical subfamily Dombeyoideae provides a particularly useful case study through which to better understand the biogeography of the WIOR, and yet its biogeography is poorly understood. Here we sampled six molecular markers from all 20 genera in the Dombeyoideae to reconstruct the most complete phylogeny to date for the subfamily. From this, divergence times, calibrated with three fossils (two dombeyoid, one malvoid), and ancestral range estimations were hypothesized. Biogeographic stochastic mapping (BSM) analyses on the maximum clade credibility tree were completed and compared to BSM analyses on 1,000 trees randomly sampled from the posterior distribution of trees resulting from the dating analysis. We found the Dombeyoideae crown node diverged ca. 53 million years ago out of a broad ancestral range involving all three major areas of its distribution: Madagascar, Africa, and Asia. The majority of diversification and dispersals in the subfamily occurred within the last ca. 10 million years, mostly from the Pliocene onwards. There were roughly five dispersals from Madagascar to Africa (and only one in reverse), at least six from Madagascar to surrounding islands of the WIOR (Mascarenes and Comoros), and one dispersal from Madagascar to Asia (and ca. 1 in reverse). Other long-distance dispersals included one from Africa to St. Helena and one from Africa to Australasia, both from within the most widespread clade, the Cheirolaena & allies clade, and one dispersal from Asia to Africa. Critically, the Dombeyoideae provide strong evidence for considering the island of Madagascar as a source for the colonization of continents, as well as the surrounding islands of the WIOR. Furthermore, narrow sympatry was a key process in the evolution of the subfamily, particularly in Madagascar and the Mascarenes.

Re-description of the early Triassic diapsid Palacrodon from the lower Fremouw formation of Antarctica.

The rapid radiation and dispersal of crown reptiles following the end-Permian mass extinction characterizes the earliest phase of the Mesozoic. Phylogenetically, this early radiation is difficult to interpret, with polytomies near the crown node, long ghost lineages, and enigmatic origins for crown group clades. Better understanding of poorly known taxa from this time can aid in our understanding of this radiation and Permo-Triassic ecology. Here, we describe an Early Triassic specimen of the diapsid Palacrodon from the Fremouw Formation of Antarctica. While Palacrodon is known throughout the Triassic and exhibits a cosmopolitan geographic range, little is known of its evolutionary relationships. We recover Palacrodon outside of crown reptiles (Sauria) but more crownward than Youngina capensis and other late Permian diapsids. Furthermore, Palacrodon possesses anatomical features that add clarity to the evolution of the stapes within the reptilian lineage, as well as incipient adaptations for arboreality and herbivory during the earliest phases of the Permo-Triassic recovery.

Evolutionary history of the Australasian Scirtinae (Scirtidae; Coleoptera) inferred from ultraconserved elements

The Scirtidae Fleming, 1821 has been identified as one of the earliest diverging groups of Polyphagan beetles and is particularly speciose in Australia. However, very little is known about the origin of the Australian scirtids and there is a need for a robust, well-supported phylogeny to guide the genus and species descriptions and understand the relationships among taxa. In this study we carried out a phylogenetic analysis of the Australian Scirtinae Fleming, 1821, using DNA sequence data from ultraconserved elements (UCEs) and included representative taxa from New Zealand, New Caledonia, South America, South Africa and Eurasia in the analysis. Bayesian analyses of a concatenated dataset from 79 taxa recovered four major Southern Hemisphere groupings and two Australian–Eurasian groupings. The Veronatus group mainly consisted of genera from New Zealand, with the three Australian representatives only distantly related to each other. Relaxed molecular clock analyses, using the estimated age of the crown node of the Polyphaga for calibration, support a Gondwanan history for four of the groups of Australian Scirtinae and a northern origin for two groups. Our results highlight the value of commercially available UCEs for resolving the phylogenetic history of ancient groups of Coleoptera.

Phylogenomic discordance suggests polytomies along the backbone of the large genus Solanum.

PremiseEvolutionary studies require solid phylogenetic frameworks, but increased volumes of phylogenomic data have revealed incongruent topologies among gene trees in many organisms both between and within genomes. Some of these incongruences indicate polytomies that may remain impossible to resolve. Here we investigate the degree of gene‐tree discordance in Solanum, one of the largest flowering plant genera that includes the cultivated potato, tomato, and eggplant, as well as 24 minor crop plants.MethodsA densely sampled species‐level phylogeny of Solanum is built using unpublished and publicly available Sanger sequences comprising 60% of all accepted species (742 spp.) and nine regions (ITS, waxy, and seven plastid markers). The robustness of this topology is tested by examining a full plastome dataset with 140 species and a nuclear target‐capture dataset with 39 species of Solanum (Angiosperms353 probe set).ResultsWhile the taxonomic framework of Solanum remained stable, gene tree conflicts and discordance between phylogenetic trees generated from the target‐capture and plastome datasets were observed. The latter correspond to regions with short internodal branches, and network analysis and polytomy tests suggest the backbone is composed of three polytomies found at different evolutionary depths. The strongest area of discordance, near the crown node of Solanum, could potentially represent a hard polytomy.ConclusionsWe argue that incomplete lineage sorting due to rapid diversification is the most likely cause for these polytomies, and that embracing the uncertainty that underlies them is crucial to understand the evolution of large and rapidly radiating lineages.

The performance of outgroup-free rooting under evolutionary radiations

Tree rooting implies a temporal dimension to phylogenies. Only after defining the position of the root node is that the ancestral-descendant relationship between branches can be fully deduced. Rooting has been usually carried out by employing evolutionarily close outgroup lineages, which is a drawback when these lineages are unavailable or unknown. Alternatively, outgroup-free rooting methods were proposed, which rely on the constancy of evolutionary rates to varying degrees. In this work we analyzed the performance of two of these methods, the midpoint rooting (MPR) and the minimal ancestor deviation (MAD), in rooting topologies evolved under challenging scenarios of fast evolutionary radiations derived from empirical data, characterized by short internal branches near the crown node. Considering all branch length combinations investigated, both methods exhibited average success rates below 50%, although MAD slightly outperformed MPR. Moreover, tree balance significantly impacted the relative performance of the methods. We found that, in four-taxa unrooted trees, the outcome of whether both methodologies will correctly root the tree can be roughly predicted by two simple dimensionless metrics: the coefficient of variation of the external branch lengths, and the ratio between the internal branch length to the total sum of branch lengths, which were employed to devise a general linear model that allowed calculating the probability of correct placing the root node for any four-taxa tree. We predicted that the performance of both outgroup-free rooting methods on loci representing the placental mammal radiation ranged between 50% and 75%.

Combining molecular and geographical data to infer the phylogeny of Lamiales and its dispersal patterns in and out of the tropics

Lamiales is one of the most intractable orders of flowering plants, with several changes in family composition, and circumscription throughout history. The order is worldwide distributed, occurring in tropical forests and frozen habitats. In this study, a comprehensive phylogeny of Lamiales was reconstructed using DNA sequences. The tree was used to infer dispersal patterns, focusing on the tropics and extratropics. Molecular and species geographic data available from public repositories were combined to address both objectives. A total of 6,910 species, and 842 genera of Lamiales were sampled using the Python tool PyPHLAWD. The tree was inferred using RAxML, and recovered a monophyletic Lamiales. All 26 families were recovered as monophyletic with high support. The families Bignoniaceae, and Plantaginaceae are remarkable examples. The first emerged as monophyletic and included tribe Jacarandeae, while the later emerged as monophyletic in its sensu lato and included both the tribes Angelonieae, and Gratioleae. Distribution points for all species were retrieved from GBIF. After filtering, 1,136,425 records were retained. Species were coded as present in extratropical or tropical environments. The in and out of the tropics dispersal patterns were inferred using a maximum likelihood approach that identifies hidden rate changes. The model recovered higher rates of transition from extratropics to tropics, estimating two rates of state transitions. When ancestral states are considered, more discrete transitions from extratropics to tropics were observed. The extratropical state was also inferred for the crown node of Lamiales and old nested nodes, revealing a rare pattern of transitions to the tropics throughout the upper Cretaceous and Tertiary. A significant phylogenetic signal was recovered for the in and out of the tropics dispersal patterns, showing that state transitions are not frequent enough to erase the effect of tree structure on the data.

Phylogeny and historical biogeography of the pantropical genus Parkia (Leguminosae, Caesalpinioideae, mimosoid clade)

Parkia R.Br. (Leguminosae, Caesalpinioideae, mimosoid clade) is a pantropical genus with approximately 35 recognized species in three taxonomic sections (Parkia, Platyparkia and Sphaeroparkia), distributed widely in tropical forests and savannas in South and Central America, Africa-Madagascar and the Indo-Pacific region. In this study, phylogenetic analyses (Maximum Likelihood and Bayesian Inference), ancestral area and habitat estimations were performed using chloroplast (matK, trnL, psbA-trnH and rps16-trnQ) and nuclear (ITS/18S/26S) DNA sequences for the purpose of testing the monophyly of Parkia and inferring the geographic origin of the genus and times of divergence of the various lineages. This enabled investigation of factors that may have influenced its diversification in both hemispheres. Our results support the monophyly of the genus. A fossil-calibrated Bayesian analysis dated the Parkia crown node to the Miocene (at c. 18.85 Ma). Biogeographic analysis reconstructed an origin in the lowlands rainforests (terra firme) in Amazonia with subsequent radiation in the Neotropical region from the Miocene onwards, with dispersion events as far as Central America, and the Atlantic Forest and the cerrado of Brazil. A single dispersion from the Neotropics to the Paleotropics is hypothesised, with subsequent smaller radiations in Africa-Madagascar and the Indo-Pacific (crown ages 3.79 and 5.15 Ma respectively). Factors that may have influenced the radiation and speciation of Parkia include the elevation of the Andes (especially in the Miocene), and more recently the closing of the Panama gap in Neotropics, the climatic fluctuations of the Pleistocene influenced the diversification of species on both continents. The elevation of the Sunda Shelf in Indo-Pacific region during the last glacial maximum (LGM) appears to be the main driving force for speciation in that region. In Africa, the low number of species may be related to extinction processes.

Phylogeny, biogeography and systematics of Dysphanieae (Amaranthaceae)

AbstractAfter a rather turbulent taxonomic history, Dysphanieae (Chenopodioideae, Amaranthaceae) were established to contain five genera, four of which are monospecific (Cycloloma, Neomonolepis, Suckleya, Teloxys) and geographically restricted, and the fifth genus, Dysphania, having a nearly worldwide distribution and comprising ca. 50 species. This study investigates the phylogeny, biogeography and taxonomy of Dysphanieae. We studied specimens from 32 herbaria to infer morphological differences and distribution areas of the species and sampled 121 accessions representing 39 accepted species of the tribe for molecular phylogenetic analyses. The molecular phylogeny tested generic relationships of the tribe and infrageneric relationships of Dysphania on the basis of two plastid DNA markers (atpB‐rbcL spacer, rpl16 intron) and two nuclear ribosomal markers (ETS, ITS) and was also used for an ancestral area reconstruction with BioGeoBEARS. Three of the monospecific genera (Neomonolepis, Suckleya, Teloxys) form a basal grade and appear to be relictual lineages of the tribe, while Cycloloma is nested within Dysphania. The ancestral area reconstruction favors a widespread ancestry for Dysphanieae, and the relictual lineages in Asia (Teloxys) and North America (Neomonolepis, Suckleya) might be explained by a wide distribution across Beringia during the Late Oligocene/Early Miocene. Dysphania likely originated in North America; however, the simultaneous diversification into three major clades, an Asian/African, an American and an Australian/African clade, indicates a widespread ancestor at the crown node of Dysphania. Our taxonomic revision results in four accepted genera in Dysphanieae, Dysphania, Neomonolepis, Suckleya and Teloxys. The sectional subdivision for Dysphania is revised. We subdivide the genus into five sections, D. sect. Adenois (13 spp.), D. sect. Botryoides (10 spp.), D. sect. Dysphania (17 spp.), D. sect. Incisa (2 spp.) and D. sect. Margaritaria (4 spp.); three strongly deviating species remain unplaced and need further attention.

Assessing the sensitivity of divergence time estimates to locus sampling, calibration points, and model priors in a RAD‐seq phylogeny ofCarexsectionSchoenoxiphium

AbstractRestriction site‐associated DNA sequencing (RAD‐seq) and related methods have become relatively common approaches to resolve species‐level phylogeny. It is not clear, however, whether RAD‐seq data matrices are well suited to relaxed clock inference of divergence times, given the size of the matrices and the abundance of missing data. We investigated the sensitivity of Bayesian relaxed clock estimates of divergence times to alternative analytical decisions on an empirical RAD‐seq phylogenetic matrix. We explored the relative contribution of secondary calibration strategies, amount of missing data, and the data partition analyzed to overall variance in divergence times inferred using BEAST MCMC analyses ofCarexsectionSchoenoxiphium(Cyperaceae)—a recent radiation for which we have nearly complete species sampling of RAD‐seq data. The crown node forSchoenoxiphiumwas estimated to be 15.22 (9.56–21.18) Ma using a single calibration point and low missing data, 11.93 (8.07–16.03) Ma using multiple calibration points and low missing data, and 8.34 (5.41–11.22) using multiple calibrations but high missing data. We found that using matrices with more than half of the individuals with missing data inferred younger mean ages for all nodes. Moreover, we have found that our molecular clock estimates are sensitive to the positions of the calibration(s) in our phylogenetic tree (using matrices with low missing data), especially when only a single calibration was applied to estimate divergence times. These results argue for sensitivity analyses and caution in interpreting divergence time estimates from RAD‐seq data.

A phylogenomic approach resolves the backbone of Prunus (Rosaceae) and identifies signals of hybridization and allopolyploidy

The genus Prunus, which contains 250–400 species, has ample genomic resources for the economically important taxa in the group including cherries, peaches, and almonds. However, the backbone of Prunus, specifically the position of the racemose group relative to the solitary and corymbose groups, remains phylogenetically uncertain. Surprisingly, phylogenomic analyses to resolve relationships in the genus are lacking. Here, we assemble transcriptomes from 17 Prunus species representing four subgenera, and use existing transcriptome assemblies, to resolve key relationships in the genus using a phylogenomic approach. From the transcriptomes, we constructed 21-taxon datasets of putatively single-copy nuclear genes with 591 and 379 genes, depending on taxon-occupancy filtering. Plastome sequences were obtained or assembled for all species present in the nuclear data set.The backbone of Prunus was resolved consistently in the nuclear and chloroplast phylogenies, but we found substantial cytonuclear discord within subgenera. Our nuclear phylogeny recovered a monophyletic racemose group, contrasting with previous studies finding paraphyly that suggests repeated allopolyploidy early in the evolutionary history of the genus. However, we detected multiple species with histories consistent with hybridization and allopolyploidy, including a deep hybridization event involving subgenus Amygdalus and the Armeniaca clade in subgenus Prunus. Analyses of gene tree conflict revealed substantial discord at several nodes, including the crown node of the racemose group. Alternative gene tree topologies that conflicted with the species tree were consistent with a paraphyletic racemose group, highlighting the complex reticulated evolutionary history of this group.

<p><strong><em>Leveillula guilanensis</em> (Erysiphales, Ascomycota): a new record from Europe with notes on the biogeography of the genus</strong></p>

Powdery mildews are the subject of intense research due to their negative impact on agriculture or horticulture. The genetic diversity of these ascomycete fungi has only been surveyed in the last decades to investigate phylogenetic relationships and species boundaries, to propose new taxa, and to assess their biogeographic history. However, many of the resulting studies are revealing only a partial picture because of the uneven species sampling across the globe. Here, we studied three Leveillula collections occurring on wild plant species in the Iberian Peninsula, a region which lacked any previous knowledge on the genus’ genetic diversity. Our study provides the first record of L. guilanensis in Europe based on specimens from two distant localities in the Iberian Peninsula; the species was so far known only from Iran. It grew on living Chondrilla juncea and was parasitized by the hyperparasite fungus Ampelomyces quisqualis. The morphological and phylogenetic assignment of another collection to L. lanuginosa is reported as well. Furthermore, we inferred a chronogram based on an Erysiphales nrITS substitution rate that dated the crown node of Leveillula back to 22.48 million years ago. Clades including the Iberian Peninsula samples originated and diversified through the Neogene (Miocene to Pleistocene). These results suggest an overall older time frame for the evolution of Leveillula compared to previous works. All in all, the present study provides additional avenues for discussing the evolutionary and biogeographic history of this genus of powdery mildews.

Presence of Air Cavities in Pods of an Endemic Legume Crotalaria pusilla Dc. (Leguminosae, Papilionoideae)

The legume lineage evolved approximately 60 Mya in the region to the north of the Tethys Sea, a shallow stretch of salt water that separated the two main groups of land masses 1 . Since then legumes have adopted various mechanisms for dispersal and colonization, among which one of the probable methods is the diaspore (a dispersal unit consisting of a seed with additional tissues that assist dispersal). The papilionoid crown node is dated 58.6 Mya with further divergence into genistoid and dalbergoid, the former dated at 56.4 Mya (ref. 1). Since their divergence, both the lineages have adopted radically different approaches to life from their common ancestors. One of the approaches is seed dispersal, which plays a central role in ecology dynamics, thereby understanding and predicting the population and community change 2 . Understanding the convergent evolution of seed dispersal is a central question in evolutionary ecology, because dispersal traits are linked to plant survival strategies and are a reflection of the reproductive assurance theory 3 .

Major lineages of Loasaceae subfam. Loasoideae diversified during the Andean uplift

The Loasoideae is the largest clade in the Loasaceae. This subfamily is widespread throughout the Neotropics and centered in the Andes, presenting an excellent opportunity to study diversification across much of temperate and mid to high-elevation areas of South America. Despite that, no studies have addressed the historical biogeography of the Loasoideae to date, leaving an important knowledge gap in this plant group. Here, we used four plastid markers (i.e., trnL–trnF, matK, trnS–trnG, and rps16) and sequenced 170 accessions (134 ingroup taxa) to infer the phylogeny of Loasoideae. We then used this phylogeny as basis to estimate divergence times using an uncorrelated relaxed molecular clock approach and seven fossils as primary calibration points. We employed the Dispersal-Extinction-Cladogenesis (DEC) approach to reconstruct the ancestral ranges of the subfamily. Our results indicate that stem Loasoideae diverged from its sister group in the Late Cretaceous to Early Paleocene (ca. 83–62 Ma). The crown node of the whole clade goes back to the Middle Paleocene to Middle Eocene (ca. 60–45 Ma), corresponding to the earliest diversification events of the extant groups, prior to most of the Andean orogeny and roughly coinciding with the Paleocene-Eocene Thermal Maximum. On the other hand, the crown nodes of most genera appear to have originated in the Oligocene and Miocene (median ages: 28–10 Ma). The diversification of some extant lineages appears to have happened in parallel to Andean uplift pulses that seem to have had an effect on the orogeny and concomitant establishment of new habitats and latitudinal corridors. The most likely ancestral areas retrieved for crown Loasoideae, are the tropical Andes and Pacific arid coast. Most of the extant clades have remained restricted to their ancestral areas. Transoceanic Long Distance Dispersal appears to have been involved in the arrival of Loasoid ancestors to South America, and in the distribution of the small clades Kissenia in Africa and Plakothira on the Marquesas Archipelago. The results presented here suggest that the historical biogeography of the continental scale radiation of Loasoideae, follows the sequence and timing of the development of temperate and mid to high-elevation habitats across South America during the Tertiary.

Fossil palm fruits from India indicate a Cretaceous origin of Arecaceae tribe Borasseae

The fossil record of palms (Arecaceae) is essential for understanding the deep evolutionary and geographical history of the family. We studied palm fruit fossils from the c. 67–64-Myr-old Deccan Intertrappean Beds of India to infer the systematic relationships of the fossils and their relevance to palm evolution. Using X-ray micro-computed tomography, physical sectioning techniques and a total-evidence phylogenetic analysis, we show that these fossils represent a crown group member of subtribe Hyphaeninae (tribe Borasseae, subfamily Coryphoideae) allied with the extant genera Satranala and Bismarckia, now endemic to Madagascar. These fossils, synonymized here as Hyphaeneocarpon indicum, provide evidence for the existence of crown group Hyphaeninae during the late Maastrichtian–early Danian. This pre-dates prior age estimates for the Hyphaeninae crown node by nearly 40 Myr and implies an earlier divergence of Borasseae. The presence of Hyphaeneocarpon in India shows that Borasseae have persisted in the Indian Ocean region for > 64 Myr. This study illustrates the utility of palm fruit characters for placing fossils in a phylogenetic context and has important implications for understanding the evolution and diversification of Borasseae and the palaeobiogeography of palms.