An evolutionary disruption of the buzz pollination syndrome in neotropical montane plants.

PREMISE: Under pollinator limitations, specialized pollination syndromes may evolve toward contrasting responses: a generalized syndrome with increased pollinator attraction, pollinator reward, and pollen transfer capacity; or the selfing syndrome with increased self-pollen deposition, but reduced pollinator attraction and pollen transfer capacity. The buzz-pollination syndrome is specialized to explore female vibrating bees as pollinators. However, vibrating bees become less-active pollinators at montane areas of the Atlantic Forest (AF) domain. This study investigated whether the specialized buzz-pollination syndrome would evolve toward an alternative floral syndrome in montane areas of the AF domain, considering a generalized and the selfing syndromes as alternative responses. METHODS: We utilized a lineage within the buzz-pollinated Miconia as study system, contrasting floral traits between montane AF-endemic and non-endemic species. We measured and validated floral traits that were proxies for pollinator attraction, reward access, pollen transfer capacity, and self-pollen deposition. We inferred the evolution of floral trait via phylogenetic comparative methods. RESULTS: AF-endemic species have selectively evolved greater reward access and more frequently had generalist pollination. Nonetheless, AF-endemic species also have selectively evolved toward lower pollen transfer capacity and greater self pollination. These patterns indicated a complex evolutionary process that has jointly favored a generalized and the selfing syndromes. CONCLUSIONS: The buzz pollination syndrome can undergo an evolutionary disruption in montane areas of the AF domain. This floral syndrome is likely more labile than often assumed, allowing buzz-pollinated plants to reproduce in environments where vibrating bees are less-reliable pollinators.

The sequential direct and indirect effects of mountain uplift, climatic niche and floral trait evolution on diversification dynamics in an Andean plant clade.

Why and how organismal lineages radiate is commonly studied through either assessing abiotic factors (biogeography, geomorphological processes, climate) or biotic factors (traits, interactions). Despite increasing awareness that both abiotic and biotic processes may have important joint effects on diversification dynamics, few attempts have been made to quantify the relative importance and timing of these factors, and their potentially interlinked direct and indirect effects, on lineage diversification. We here combine assessments of historical biogeography, geomorphology, climatic niche, vegetative and floral trait evolution to test whether these factors jointly, or in isolation, explain diversification dynamics of a Neotropical plant clade (Merianieae, Melastomataceae). After estimating ancestral areas and the changes in niche and trait disparity over time, we employ Phylogenetic Path Analyses as a synthesis tool to test eleven hypotheses on the individual direct and indirect effects of these factors on diversification rates. We find strongest support for interlinked effects of colonization of the uplifting Andes during the mid-Miocene and rapid abiotic climatic niche evolution in explaining a burst in diversification rate in Merianieae. Within Andean habitats, later increases in floral disparity allowed for the exploitation of wider pollination niches (i.e., shifts from bee to vertebrate pollinators), but did not affect diversification rates. Our approach of including both vegetative and floral trait evolution, rare in assessments of plant diversification in general, highlights that the evolution of woody habit and larger flowers preceded the colonization of the Andes, but was likely critical in enabling the rapid radiation in montane environments. Overall, and in concert with the idea that ecological opportunity is a key element of evolutionary radiations, our results suggest that a combination of rapid niche evolution and trait shifts were critical for the exploitation of newly available niche space in the Andes in the mid-Miocene. Further, our results emphasize the importance of incorporating both abiotic and biotic factors into the same analytical framework if we aim to quantify the relative and interlinked effects of these processes on diversification.

An updated and extended version of the Melastomataceae probe set for target capture.

Premise: A probe set was previously designed to target 384 nuclear loci in the Melastomataceae family; however, when trying to use it, we encountered several practical and conceptual problems, such as the presence of sequences in reverse complement, intronic regions with stop codons, and other issues. This raised concerns regarding the use of this probe set for sequence recovery in Melastomataceae. Methods: In order to correct these issues, we cleaned the Melastomataceae probe set, extended it with additional sequences, and compared its performance with the original version. Results: The final probe set targets 396 putative nuclear loci represented by 6009 template sequences. The probe set has been made available, along with details on the cleaning process, for reproducibility. We show that the new probe set performs better than the original version in terms of sequence recovery. Discussion: This updated, extended, and cleaned probe set will improve the availability of phylogenomic resources across the Melastomataceae family. It is fully compatible with sequence recovery and extraction pipelines. The cleaning process can also be applied to any plant-targeting probe set that would need to be cleaned or updated if new genomic resources for the targeted taxa become available.

Out of chaos: Phylogenomics of Asian Sonerileae.

Sonerileae is a diverse Melastomataceae lineage comprising ca. 1000 species in 44 genera, with >70% of genera and species distributed in Asia. Asian Sonerileae are taxonomically intractable with obscure generic circumscriptions. The backbone phylogeny of this group remains poorly resolved, possibly due to complexity caused by rapid species radiation in early and middle Miocene, which hampers further systematic study. Here, we used genome resequencing data to reconstruct the phylogeny of Asian Sonerileae. Three parallel datasets, viz. single-copy ortholog (SCO), genomic SNPs, and whole plastome, were assembled from genome resequencing data of 205 species for this purpose. Based on these genome-scale data, we provided the first well resolved phylogeny of Asian Sonerileae, with 34 major clades identified and 74% of the interclade relationships consistently resolved by both SCO and genomic data. Meanwhile, widespread phylogenetic discordance was detected among SCO gene trees as well as species trees reconstructed using different tree estimation methods (concatenation/site-based coalescent method/summary method) or different datasets (SCO/genomic/plastome). We explored sources of discordance using multiple approaches and found that the observed discordance in Asian Sonerileae was mainly caused by a combination of biased distribution of missing data, random noise from uninformative genes, incomplete lineage sorting, and hybridization/introgression. Exploration of these sources can enable us to generate hypotheses for future testing, which is the first step towards understanding the evolution of Asian Sonerileae. We also detected high levels of homoplasy for some characters traditionally used in taxonomy, which explains current chaotic generic delimitations. The backbone phylogeny of Asian Sonerileae revealed in this study offers a solid basis for future taxonomic revision at the generic level.

Beneath a hairy problem: Phylogeny, morphology, and biogeography circumscribe the new Miconia supersection Discolores (Melastomataceae: Miconieae).

Miconia is among the largest plant genera in the Neotropics and a taxonomically complex lineage. Indeed, molecular phylogenetic data shows that none of its traditionally accepted sections are monophyletic, preventing taxonomic advances within the genus. Miconia is the largest plant genus in the Brazilian Atlantic Forest, including three main lineages, the Leandra s.s. clade (ca. 215 spp.), the Miconia sect. Chaenanthera (24 spp.), and the Miconia discolor clade (estimated 77 spp.). Out of these lineages, the Miconia discolor clade is the only currently lacking phylogenetic data, complicating its taxonomy. In this study, we reconstruct the phylogeny of the Miconia discolor clade, using three plastid (atpF-H, psbK-I, and psaI-accD) and two nuclear (ETS and ITS) markers. We sampled 60 out of the 77 species of the group, representing 78% of its diversity. Taxa were selected considering their distribution, morphology, and previous phylogenetic knowledge. We used the newly reconstructed phylogeny to better understand phylogenetic relationships among Atlantic Forest species and morphologically similar taxa, and to propose a new infrageneric classification for the Miconia discolor clade: the Miconia supersection Discolores. We further studied the evolution of seven morphological characters using a Maximum Likelihood approach, and estimated the ancestral range distribution of various lineages in order to understand the biogeographic history of this clade. We found that dichasial inflorescences represent the ancestral condition within Miconia, subsequently giving rise to scorpioid and glomerulate inflorescences in the studied group. We describe Miconia supersect. Discolores, originated in the Amazon region, which is recognized by a dense layer of branched tricomes covering young branches and non-dichasial inflorescences, including three main lineages: (i) Miconia sect. Albicantes, characterized by persistent bracts and arachnoid indument on the abaxial surface of leaves, mainly distributed in the Amazon basin; (ii) Miconia sect. Discolores, characterized by caducous calyx lobes and glomerulate inflorescences, centered in the Atlantic Forest; and (iii) Miconia sect. Multispicatae, characterized by leaves not completely covered with indument, and capitate stigma, mainly distributed in the Atlantic Forest. All three sections and the supersection originated in the Neogene, between the Late Miocene and the Early Pliocene.

A multidisciplinary framework for biodiversity prediction in the Brazilian Atlantic Forest hotspot / Integrando disciplinas para a predição da biodiversidade da Floresta Atlântica no Brasil

Abstract: We briefly describe selected results from our thematic project focused on the biodiversity of the Atlantic Forest ("AF BIOTA"), which was jointly funded by FAPESP's BIOTA Program, the U.S. National Science Foundation Dimensions of Biodiversity Program, and the National Aeronautics and Space Administration (NASA). As one of the five most important hotspots of biodiversity in the world, the Atlantic Forest (AF) holds less than 16% of its vegetation cover, yet, amongst the hotspots, it still harbors one of the highest numbers of species, including endemics. By gathering specialists across multiple disciplines (biology, geology, engineering), we aimed to understand how this megabiodiversity was built through time, informing biodiversity science and conservation. Among the results, we trained 18 Master's and 26 Ph.D. students, published more than 400 peer-reviewed papers that improved our knowledge about the forest's biologic and climatic diversity and dynamics through time, developed new analytical methods, produced outreach videos and articles, and provided data to help define biodiversity conservation policies.

Resumo: Descrevemos de forma resumida resultados selecionados do nosso projeto temático com foco na biodiversidade da Floresta Atlântica ("AF BIOTA"), que foi financiado pelo BIOTA FAPESP e pelo programa "Dimensions of Biodiversity" da "U.S. National Science Foundation" e "National Aeronautics and Space Administration" (NASA). Devido à sua megabiodiversidade (que inclui várias espécies endêmicas), e por restar menos de 16% da vegetação original, a Floresta Atlântica (FA) é uma das cinco áreas mais importantes para a biodiversidade do planeta ("biodiversity hotspot"). Reunimos especialistas de diversas disciplinas (biologia, geologia, engenharia) visando compreender como essa megabiodiversidade evoluiu ao longo do tempo e fornecer informações científicas para a sua conservação. Dentre os resultados obtidos, nós formamos 18 mestres e 26 doutores, publicamos mais de 400 artigos científicos que aumentaram o conhecimento sobre a diversidade biológica e climática da FA e sua dinâmica ao longo do tempo, desenvolvemos novos métodos analíticos, produzimos material de divulgação científica e fornecemos dados para desenvolver políticas públicas de conservação da biodiversidade.

A nuclear phylogenomic study of the angiosperm order Myrtales, exploring the potential and limitations of the universal Angiosperms353 probe set.

PREMISE: To further advance the understanding of the species-rich, economically and ecologically important angiosperm order Myrtales in the rosid clade, comprising nine families, approximately 400 genera and almost 14,000 species occurring on all continents (except Antarctica), we tested the Angiosperms353 probe kit. METHODS: We combined high-throughput sequencing and target enrichment with the Angiosperms353 probe kit to evaluate a sample of 485 species across 305 genera (76% of all genera in the order). RESULTS: Results provide the most comprehensive phylogenetic hypothesis for the order to date. Relationships at all ranks, such as the relationship of the early-diverging families, often reflect previous studies, but gene conflict is evident, and relationships previously found to be uncertain often remain so. Technical considerations for processing HTS data are also discussed. CONCLUSIONS: High-throughput sequencing and the Angiosperms353 probe kit are powerful tools for phylogenomic analysis, but better understanding of the genetic data available is required to identify genes and gene trees that account for likely incomplete lineage sorting and/or hybridization events.

Low bee visitation rates explain pollinator shifts to vertebrates in tropical mountains.

Evolutionary shifts from bee to vertebrate pollination are common in tropical mountains. Reduction in bee pollination efficiency under adverse montane weather conditions was proposed to drive these shifts. Although pollinator shifts are central to the evolution and diversification of angiosperms, we lack experimental evidence of the ecological processes underlying such shifts. Here, we combine phylogenetic and distributional data for 138 species of the Neotropical plant tribe Merianieae (Melastomataceae) with pollinator observations of 11 and field pollination experiments of six species to test whether the mountain environment may indeed drive such shifts. We demonstrate that shifts from bee to vertebrate pollination coincided with occurrence at high elevations. We show that vertebrates were highly efficient pollinators even under the harsh environmental conditions of tropical mountains, whereas bee pollination efficiency was lowered significantly through reductions in flower visitation rates. Furthermore, we show that pollinator shifts in Merianieae coincided with the final phases of the Andean uplift and were contingent on adaptive floral trait changes to alternative rewards and mechanisms facilitating pollen dispersal. Our results provide evidence that abiotic environmental conditions (i.e. mountain climate) may indeed reduce the efficiency of a plant clade's ancestral pollinator group and correlate with shifts to more efficient new pollinators.

The Herbarium 2021 Half-Earth Challenge Dataset and Machine Learning Competition.

Herbarium sheets present a unique view of the world's botanical history, evolution, and biodiversity. This makes them an all-important data source for botanical research. With the increased digitization of herbaria worldwide and advances in the domain of fine-grained visual classification which can facilitate automatic identification of herbarium specimen images, there are many opportunities for supporting and expanding research in this field. However, existing datasets are either too small, or not diverse enough, in terms of represented taxa, geographic distribution, and imaging protocols. Furthermore, aggregating datasets is difficult as taxa are recognized under a multitude of names and must be aligned to a common reference. We introduce the Herbarium 2021 Half-Earth dataset: the largest and most diverse dataset of herbarium specimen images, to date, for automatic taxon recognition. We also present the results of the Herbarium 2021 Half-Earth challenge, a competition that was part of the Eighth Workshop on Fine-Grained Visual Categorization (FGVC8) and hosted by Kaggle to encourage the development of models to automatically identify taxa from herbarium sheet images.

An algorithm competition for automatic species identification from herbarium specimens.

PREMISE: Plant biodiversity is threatened, yet many species remain undescribed. It is estimated that >50% of undescribed species have already been collected and are awaiting discovery in herbaria. Robust automatic species identification algorithms using machine learning could accelerate species discovery. METHODS: To encourage the development of an automatic species identification algorithm, we submitted our Herbarium 2019 data set to the Fine-Grained Visual Categorization sub-competition (FGVC6) hosted on the Kaggle platform. We chose to focus on the flowering plant family Melastomataceae because we have a large collection of imaged herbarium specimens (46,469 specimens representing 683 species) and taxonomic expertise in the family. As is common for herbarium collections, some species in this data set are represented by few specimens and others by many. RESULTS: In less than three months, the FGVC6 Herbarium 2019 Challenge drew 22 teams who entered 254 models for Melastomataceae species identification. The four best algorithms identified species with >88% accuracy. DISCUSSION: The FGVC competitions provide a unique opportunity for computer vision and machine learning experts to address difficult species-recognition problems. The Herbarium 2019 Challenge brought together a novel combination of collections resources, taxonomic expertise, and collaboration between botanists and computer scientists.

A two-tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae.

PREMISE: Putatively single-copy nuclear (SCN) loci, which are identified using genomic resources of closely related species, are ideal for phylogenomic inference. However, suitable genomic resources are not available for many clades, including Melastomataceae. We introduce a versatile approach to identify SCN loci for clades with few genomic resources and use it to develop probes for target enrichment in the distantly related Memecylon and Tibouchina (Melastomataceae). METHODS: We present a two-tiered pipeline. First, we identified putatively SCN loci using MarkerMiner and transcriptomes from distantly related species in Melastomataceae. Published loci and genes of functional significance were then added (384 total loci). Second, using HybPiper, we retrieved 689 homologous template sequences for these loci using genome-skimming data from within the focal clades. RESULTS: We sequenced 193 loci common to Memecylon and Tibouchina. Probes designed from 56 template sequences successfully targeted sequences in both clades. Probes designed from genome-skimming data within a focal clade were more successful than probes designed from other sources. DISCUSSION: Our pipeline successfully identified and targeted SCN loci in Memecylon and Tibouchina, enabling phylogenomic studies in both clades and potentially across Melastomataceae. This pipeline could be easily applied to other clades with few genomic resources.

Flower morphology is correlated with distribution and phylogeny in Bertolonia (Melastomataceae), an herbaceous genus endemic to the Atlantic Forest.

Several centers of endemism have been proposed for Melastomataceae, particularly in Amazonia and Atlantic Forest. Despite the high degree of human-caused degradation in the last 500 years, the Atlantic Forest still presents some of the largest levels of diversity and endemism across all angiosperms. With several recently described species in the last decade, the knowledge on Bertolonia's distribution and morphological characterization has changed, with most new species found in northern Atlantic Forest and with different flower color patterns than the species from southern Atlantic Forest. We first tested the monophyly of the genus sampling over 85% of its species to generate a reliable phylogenetic hypothesis. Afterwards, we used Bertolonia as a model group to study distribution patterns and morphological evolution of lineages in the Atlantic Forest. Bertolonia is particularly interesting to address such questions because it is endemic to this domain, with species distributed either in the southern, central or northern portions of the Atlantic Forest. The second step of our work aimed to respond (1) Do closely related species endemic to Atlantic Forest tend to have similar distributions and share similar morphological traits? and (2) Are floral traits more conserved within clades than vegetative characters? We hypothesize that both (1) and (2) are true due to the low dispersal ability and consequent microendemic distribution of most species in the genus. Our results confirm the veracity of the proposed hypotheses. Three major groups were recovered in our analysis: marmorata, formosa and nymphaeifolia clades. Most species that occur in northern Atlantic Forest were recovered in the marmorata clade. Most species distributed in central Atlantic Forest were recovered in the formosa clade, and the ones from the southern Atlantic Forest in the nymphaeifolia clade. A similar pattern was recovered with reproductive characters reconstructed across the phylogeny. Generally, species with pink petals and cream-colored anthers with an extrorse pore occur in northern Atlantic Forest, and species with white petals and yellow anthers with an introrse or apical pore tend to occur in southern Atlantic Forest. Some vegetative characters also have evolutionary congruence and are restricted to one or few lineages, while other characters, such as the texture of the leaf blade surface, have a strong taxonomic value and are useful to identify species, but are not homologous. Our analysis also indicates that the division between southern and northern Atlantic Forest could also be related to ancient events, not only linked with recent phylogeographic patterns. Moreover, we suggest that the orientation of the anther pore in Bertolonia could be related with diversity of species and stability of its populations. In summary, we corroborate, based on the evolutionary history of Bertolonia, that closely related species endemic to Atlantic Forest tend to have similar distributions and share similar morphological floral traits.

Miconia lucenae (Melastomataceae), a new species from montane Atlantic Forest in Espírito Santo, Brazil.

We describe Miconia lucenae R.Goldenb. & Michelang., a new species from the montane Atlantic Forest in Santa Teresa in the state of Espírito Santo. Our analysis, based on three plastid spacers (atpF-atpH, psbK-psbl and trnS-trnG), one plastid gene (ndhF, not available for M. lucenae), and two nuclear ribosomal loci (nrITS and nrETS), showed that it belongs to a small clade with Miconia paradoxa (Mart. ex DC.) Triana (Minas Gerais) and M. michelangeliana R.Goldenb. & L.Kollmann (Espírito Santo). The three species in the "Paradoxa clade" can be recognized by the plants with glabrous or glabrescent branches and leaves, white petals and yellow stamens, these with the connectives not prolonged below the thecae, ventrally unappendaged, dorsally unappendaged or with a minute tooth, the latter bilobed or not, glabrous ovary, and the fruits with a persistent calyx. Miconia lucenae can be recognized, among the species in this clade, by the shrubby plants with terete young branches, short inflorescences, usually with red axes, and the 2-bracteolate, sessile, 4-merous flowers, with a ciliolate inner portion of the sepals, lanceolate petals, and 4-celled ovaries. This species can be considered as endangered according to IUCN criteria.

Modularity increases rate of floral evolution and adaptive success for functionally specialized pollination systems.

Angiosperm flowers have diversified in adaptation to pollinators, but are also shaped by developmental and genetic histories. The relative importance of these factors in structuring floral diversity remains unknown. We assess the effects of development, function and evolutionary history by testing competing hypotheses on floral modularity and shape evolution in Merianieae (Melastomataceae). Merianieae are characterized by different pollinator selection regimes and a developmental constraint: tubular anthers adapted to specialized buzz-pollination. Our analyses of tomography-based 3-dimensional flower models show that pollinators selected for functional modules across developmental units and that patterns of floral modularity changed during pollinator shifts. Further, we show that modularity was crucial for Merianieae to overcome the constraint of their tubular anthers through increased rates of evolution in other flower parts. We conclude that modularity may be key to the adaptive success of functionally specialized pollination systems by making flowers flexible (evolvable) for adaptation to changing selection regimes.

Erratum: Author Correction: Modularity increases rate of floral evolution and adaptive success for functionally specialized pollination systems.

[This corrects the article DOI: 10.1038/s42003-019-0697-7.].

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.

Real-time PCR detection of a 16S rRNA single mutation of Helicobacter pylori isolates associated with reduced susceptibility and resistance to tetracycline in the gastroesophageal mucosa of individual hosts.

The molecular mechanism of Helicobacter pylori resistance to tetracycline involves mutations in the primary binding site of the ribosome. A resistance or reduced susceptibility to tetracycline could be the result of single, double or triple mutations in the 16S rRNA gene of H. pylori. We investigated if the genotype was correlated to tetracycline resistance as determined phenotypically in vitro for 96 H. pylori isolates in the gastroesophageal mucosa of Venezuelan individual hosts. E-test for antimicrobial susceptibility test and real-time PCR for the detection of 16S rRNA gene mutations were performed in 96 H. pylori isolates (48 obtained from antrum, and 48 from oesophagus) from eight dyspeptic patients. In the gastric mucosa, 38 isolates were identified sensitive and 10 resistant to tetracycline by E-test, whereas 44 sensitive and 4 resistant isolates were found in the oesophagus. Real-time PCR detection of the 16S rRNA gene exhibited mutants with a single base-pair substitution (AGA926GGA) in six antrum isolates and seven oesophagus isolates, whereas only three harboured a low level of tetracycline resistance in vitro. Our results indicate that real-time PCR detection of 16S rRNA is a reliable method to classify among tetracycline-resistant genotypes and useful in patients who have experienced a first-line treatment failure with triple therapy.

Imaging the unimaginable: Medical imaging in the realm of photography.

There is an almost innate urge in human beings to represent reality in a visual form. From rock art in the Paleolithic to images of galaxies, the quotidian and the extraordinary have been visually represented through the ages. Medical and scientific disciplines are no exception. Accurate representation of the human body structures and anatomy based on cadaver dissections was almost not possible up to the Renaissance due to ethical, social, and religious beliefs and objections. The works of Leonardo da Vinci (1452-1519) and others and, later, Andreas Vesalius (1514-1564), who produced De Humanis Corporis Fabrica, are considered landmarks in the history of medicine. During the following centuries medical and scientific illustration relied upon the expertise of physician-artists and scientist-artists until a new paradigm appeared in the realm of scientific (medical) illustration: the invention of photography in the 19th century. Two of the medical disciplines most rapidly influenced by photography were dermatology and pathology, both macro- and microscopic. Physicians rapidly started to use photographs as a tool for consultation, documentation, and education, and large collections of images were amassed by individuals and institutions for these purposes. Photographic images are produced by visible light impressing a light-sensitive material such as a silver halide plate, and nowadays a silicon chip. But photons are reflected by nontransparent objects, including the human skin. Developments in science and technology allowed the use of other types of radiation to reveal internal structures in the human body and, most interestingly, noninvasively. Thus today much of the medical diagnosis and treatment is guided by the so-called medical imaging with the use of these techniques, that is, medical photography, endoscopy, x-ray radiography, computer-aided tomography, magnetic resonance imaging, ultrasonography, thermography, and nuclear medicine functional imaging techniques as positron emission tomography (PET) and single-photon emission computed tomography (SPECT). Some of these techniques are being applied at the microscopic level to study cell structure and even functional changes in real time. All these advancements in science and technology applied to medicine and other disciplines pose the question as to what extent physicians are trading their capabilities as clinicians. Ethics issues add to the complexity of this new era governed by constant changes in scientific paradigms.

Beyond buzz-pollination - departures from an adaptive plateau lead to new pollination syndromes.

Pollination syndromes describe recurring adaptation to selection imposed by distinct pollinators. We tested for pollination syndromes in Merianieae (Melastomataceae), which contain bee- (buzz-), hummingbird-, flowerpiercer-, passerine-, bat- and rodent-pollinated species. Further, we explored trait changes correlated with the repeated shifts away from buzz-pollination, which represents an 'adaptive plateau' in Melastomataceae. We used random forest analyses to identify key traits associated with the different pollinators of 19 Merianieae species and estimated the pollination syndromes of 42 more species. We employed morphospace analyses to compare the morphological diversity (disparity) among syndromes. We identified three pollination syndromes ('buzz-bee', 'mixed-vertebrate' and 'passerine'), characterized by different pollen expulsion mechanisms and reward types, but not by traditional syndrome characters. Further, we found that 'efficiency' rather than 'attraction' traits were important for syndrome circumscription. Contrary to syndrome theory, our study supports the pooling of different pollinators (hummingbirds, bats, rodents and flowerpiercers) into the 'mixed-vertebrate' syndrome, and we found that disparity was highest in the 'buzz-bee' syndrome. We conclude that the highly adaptive buzz-pollination system may have prevented shifts towards classical pollination syndromes, but provided the starting point for the evolution of a novel set of distinct syndromes, all having retained multifunctional stamens that provide pollen expulsion, reward and attraction.

The norpurpureine alkaloid from Annona purpurea inhibits human platelet activation in vitro.

BACKGROUND: The leaves of Annona purpurea have yielded several alkaloids with anti-aggregation activities against rabbit platelets. This is promising in the search for agents that might act against platelets and reduce the incidence of cardiovascular diseases. Since significant differences in platelet function have been reported between human and animal platelets, a study focusing on the effect of A. purpurea extracts against human platelet activation is necessary. METHODS: The compounds in an A. purpurea ethanolic extract underwent bio-guided fractionation and were used for in vitro human platelet aggregation assays to isolate the compounds with anti-platelet activity. The bioactive compounds were identified by spectroscopic analysis. Additional platelet studies were performed to characterize their action as inhibitors of human platelet activation. RESULTS: The benzylisoquinoline alkaloid norpurpureine was identified as the major anti-platelet compound. The IC50 for norpurpureine was 80 µM against platelets when stimulated with adenosine 5'-diphosphate (ADP), collagen and thrombin. It was pharmacologically effective from 20 to 220 µM. Norpurpureine (220 µM) exhibited its in vitro effectiveness in samples from 30 healthy human donors who did not take any drugs during the 2 weeks prior to the collection. Norpurpureine also gradually inhibited granule secretion and adhesion of activated platelets to immobilized fibrinogen. At the intra-platelet level, norpurpureine prevented agonist-stimulated calcium mobilization and cAMP reduction. Structure-activity relationship analysis indicates that the lack of a methyl group at the nitrogen seems to be key in the ability of the compound to interact with its molecular target. CONCLUSION: Norpurpureine displays a promising in vitro pharmacological profile as an inhibitor of human platelet activation. Its molecular target could be a common effector between Ca2+ and cAMP signaling, such as the PLC-PKC-Ca2+ pathway and PDEs. This needs further evaluation at the protein isoform level.