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Premise: Pollinator sharing of co-flowering plants may result in interspecific pollen receipt with a fitness cost. However, the underlying factors that determine the effects of heterospecific pollen (HP) are not fully understood. Moreover, the cost of stigma closure induced by HP may be more severe for plants with special touch-sensitive stigmas than for plants with non-touch-sensitive stigmas. Very few studies have assessed HP effects on stigma behavior., Methods: We conducted hand-pollination experiments with 10 HP donors to estimate HP effects on stigma behavior and stigmatic pollen germination in Campsis radicans (Bignoniaceae) at low and high pollen loads. We assessed the role of phylogenetic distance between donor and recipient, pollen size, and pollen aperture number in mediating HP effects. Additionally, we observed pollen tube growth to determine the conspecific pollen-tube-growth advantage., Results: Stigma behavior differed significantly with HP of different species. Pollen load increased, while pollen size decreased, the percentage of permanent closure and stigmatic germination of HP. Stigmatic HP germination increased with increasing aperture number. However, HP effects did not depend on phylogenetic distance. In addition, conspecific pollen had a pollen-tube-growth advantage over HP., Conclusions: Our results provide a good basis for understanding the stigma-pollen recognition process of plant taxa with touch-sensitive stigmas. We concluded that certain flowering traits drive the HP effects on the post-pollination period. To better understand the impact of pollinator sharing and interspecific pollen transfer on plant evolution, we highlight the importance of evaluating more factors that determine HP effects at the community level., (© 2022 Botanical Society of America.)

Main Conclusion: Bignoniaceae species have conserved chloroplast structure, with hotspots of nucleotide diversity. Several genes are under positive selection, and can be targets for evolutionary studies. Bignoniaceae is one of the most species-rich family of woody plants in Neotropical seasonally dry forests. Here we report the assembly of Handroanthus impetiginosus chloroplast genome and evolutionary comparative analyses of ten Bignoniaceae species representing the genera for which whole-genome chloroplast sequences were available. The chloroplast genome of H. impetiginosus is 159,462 bp in size and has a similar structure compared to the other nine species. The total number of genes was slightly variable amongst the Bignoniaceae, ranging from 124 in H. impetiginosus to 144 in Anemopaegma acutifolium. The inverted repeat (IR) size was variable, ranging from 24,657 bp (Tecomaria capensis) to 40,481 bp (A. acutifolium), due to the contraction and retraction at its boundaries. However, gene boundaries were very similar among the ten species. We found 98 forward and palindromic dispersed repeats, and 85 simple sequence repeats (SSRs). In general, chloroplast sequences were highly conserved, with few nucleotide diversity hotspots in the genes accD, clpP, rpoA, ycf1, ycf2. The phylogenetic analysis based on 77 coding genes was highly consistent with Angiosperm Phylogeny Group (APG) IV. Our results also indicate that most genes are under negative selection or neutral evolution. We found no evidence of branch-site selection, implying that H. impetiginosus is not evolving faster than the other species analyzed, notwithstanding we found site positive selection signal in several genes. These genes can provide targets for evolutionary studies in Bignoniaceae and Lamiales species.

The mechanisms and processes underlying patterns of species distributions have intrigued ecologists and biogeographers for a long time. The Neotropics is the most species-rich region in the World, representing an excellent model for studying the drivers of diversification. In this study, we used a phylogenomic approach to infer relationships and examine the role of major geological and climatic events in shaping biogeographic patterns within Amphilophium (Bignonieae, Bignoniaceae), a genus of Neotropical lianas. Even though Amphilophium is broadly distributed across the Neotropics, it is centered in Amazonia and the Atlantic rainforest. We generated nearly-complete plastome sequences for 32 species of Amphilophium, representing 70% of the species diversity in the genus. The final dataset included 78 plastid-coding regions and was analyzed under Maximum Likelihood and Bayesian approaches to reconstruct the phylogeny of Amphilophium. We also used this dataset to estimate divergence times using a Bayesian relaxed-clock approach. We further inferred ancestral ranges, migration events, and shifts in diversification rates using a branch-specific diversification model and the Dispersal-Extinction-Cladogenesis (DEC) model implemented in a Bayesian phylogenetic framework. Overall, we obtained a well-resolved and strongly supported phylogeny for Amphilophium, with five main clades that are well characterized by morphological features. Amphilophium originated in the Early Oligocene, and started to diversify in the Late Oligocene. The first diversification event involved a split between Amazonian and Atlantic forest clades. These two clades showed very different diversification scenarios. Divergence within the Atlantic forest clade began in the Mid-Oligocene, while the Amazonian clade underwent rapid diversification starting in the Late Miocene. In-situ speciation characterized the Amazonian clade, whereas allopatric speciation driven by migration events into other Neotropical biomes were mostly inferred within the Atlantic forest clade. The diversification of Amphilophium in the Neotropics was triggered by major geological events and changes in landscape that occurred during the Late Paleogene and Neogene, with little influence of the climatic changes of the Pleistocene ice ages. The divergence times and range inferences support the role of the Western Amazonian "megawetlands" and the formation of the South American "dry diagonal" as key climatic and geological barriers that separated the Atlantic forest from the Amazonian lowlands. Timing of migration events agrees with a Mid-Miocene closure of the Central American Seaway., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Combining high-throughput sequencing data with amplicon sequences allows the reconstruction of robust phylogenies based on comprehensive sampling of characters and taxa. Here, we combine Next Generation Sequencing (NGS) and Sanger sequencing data to infer the phylogeny of the "Adenocalymma-Neojobertia" clade (Bignonieae, Bignoniaceae), a diverse lineage of Neotropical plants, using Maximum Likelihood and Bayesian approaches. We used NGS to obtain complete or nearly-complete plastomes of members of this clade, leading to a final dataset with 54 individuals, representing 44 members of ingroup and 10 outgroups. In addition, we obtained Sanger sequences of two plastid markers (ndhF and rpl32-trnL) for 44 individuals (43 ingroup and 1 outgroup) and the nuclear PepC for 64 individuals (63 ingroup and 1 outgroup). Our final dataset includes 87 individuals of members of the "Adenocalymma-Neojobertia" clade, representing 66 species (ca. 90% of the diversity), plus 11 outgroups. Plastid and nuclear datasets recovered congruent topologies and were combined. The combined analysis recovered a monophyletic "Adenocalymma-Neojobertia" clade and a paraphyletic Adenocalymma that also contained a monophyletic Neojobertia plus Pleonotoma albiflora. Relationships are strongly supported in all analyses, with most lineages within the "Adenocalymma-Neojobertia" clade receiving maximum posterior probabilities. Ancestral character state reconstructions using Bayesian approaches identified six morphological synapomorphies of clades namely, prophyll type, petiole and petiolule articulation, tendril ramification, inflorescence ramification, calyx shape, and fruit wings. Other characters such as habit, calyx cupular trichomes, corolla color, and corolla shape evolved multiple times. These characters are putatively related with the clade diversification and can be further explored in diversification studies., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Chromosome numbers and heterochromatin banding pattern variability have been shown to be useful for taxonomic and evolutionary studies of different plant taxa. Bignonieae is the largest tribe of Bignoniaceae, composed mostly by woody climber species whose taxonomies are quite complicated. We reviewed and added new data concerning chromosome numbers in Bignonieae and performed the first analyses of heterochromatin banding patterns in that tribe based on the fluorochromes chromomycin A3 (CMA) and 4'-6-diamidino-2-phenylindole (DAPI). We confirmed the predominant diploid number 2n = 40, as well as variations reported in the literature (dysploidy in Mansoa [2n = 38] and polyploidy in Dolichandra ungis-cati [2n = 80] and Pyrostegia venusta [2n = 80]). We also found a new cytotype for the genus Anemopaegma (Anemopaegma citrinum, 2n = 60) and provide the first chromosome counts for five species (Adenocalymma divaricatum, Amphilophium scabriusculum, Fridericia limae, F. subverticillata, and Xylophragma myrianthum). Heterochromatin analyses revealed only GC-rich regions, with six different arrangements of those bands. The A-type (one large and distal telomeric band) were the most common, although the presence and combinations of the other types appear to be the most promising for taxonomic studies.

Premise of the Study: Bignoniaceae is an important component of neotropical forests and a model for evolutionary and biogeographical studies. A previous combination of molecular markers and morphological traits improved the phylogeny of the group. Here we demonstrate the value of next-generation sequencing (NGS) to assemble the chloroplast genome of eight Anemopaegma species and solve taxonomic problems., Methods: Three NGS platforms were used to sequence total DNA of Anemopaegma species. After genome assembly and annotation, we compared chloroplast genomes within Anemopaegma, with other Lamiales species, and the evolutionary rates of protein-coding genes using Tanaecium tetragonolobum as the outgroup. Phylogenetic analyses of Anemopaegma with different data sets were performed., Key Results: Chloroplast genomes of Anemopaegma species ranged from 167,413 bp in A. foetidum to 168,987 bp in A. acutifolium ("typical" form). They exhibited a characteristic quadripartite structure with a large single-copy region (75,070-75,761 bp), a small single-copy region (12,766-12,817 bp) and a pair of inverted repeat regions (IRs) (39,480-40,481) encoding an identical set of 112 genes. An inversion of a fragment with ca. 8 kb, located in the IRs and containing the genes trnI-AAU, ycf2, and trnL-CAA, was observed in these chloroplast genomes when compared with those of other Lamiales., Conclusions: Anemopaegma species have the largest genomes within the Lamiales possibly due to the large amount of repetitive sequences and IR expansion. Variation was higher in coding regions than in noncoding regions, and some genes were identified as markers for differentiation between species. The use of the entire chloroplast genome gave better phylogenetic resolution of the taxonomic groups. We found that two forms of A. acutifolium result from different maternal lineages., (© 2017 Botanical Society of America.)

Numerous plant species worldwide including Palicourea marcgravii (Rubiaceae) and Tanaecium bilabiatum (Bignoniaceae) in Brazil cause acute cardiac failure (sudden death) and are known to contain monofluoroacetate (MFA). Other Bignoniaceae species including Fridericia japurensis (Arrabidaea japurensis) are reported to cause sudden death in livestock in the Brazilian state of Roraima and are suspected to contain MFA due to the similarity of clinical signs. In this study herbarium specimens of Fridericia japurensis and field collections suspected to be F. japurensis were analyzed for MFA, and plant material from the field collections was dosed to rabbits. No MFA was detected in the herbarium specimens authoritatively identified as F. japurensis; however, MFA was detected in the field collections, which were identified as T. bilabiatum. Rabbits dosed orally with T. bilabiatum died acutely. Voucher toxic specimens initially described as F. japurensis were incorrectly identified, and the correct botanical name for this plant is T. bilabiatum (Arrabidaea bilabiata). Based on this study we conclude that there are no data to support the toxicity of F. japurensis and that the plant previously reported under this name as causing acute cardiac failure in cattle in Roraima is T. bilabiatum. This research highlights the importance of voucher specimens as part of any toxic plant investigation and corrects the literature regarding these toxic plants., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study presents the most complete generic phylogenetic framework to date for the tribe Coleeae (Bignoniaceae), which is endemic to Madagascar and the other smaller islands in the western part of the Indian Ocean. The study is based on plastid and nuclear DNA regions and includes 47 species representing the five currently recognized genera (including all the species occurring in the western Indian Ocean region). Bayesian and maximum likelihood analyses supported (i) the monophyly of the tribe, (ii) the monophyly of Phylloctenium, Phyllarthron and Rhodocolea and (iii) the paraphyly of Colea due to the inclusion of species of Ophiocolea. The latter genus was also recovered paraphyletic due to the inclusion of two species of Colea (C. decora and C. labatii). The taxonomic implications of the mutual paraphyly of these two genera are discussed in light of morphological evidence, and it is concluded that the two genera should be merged, and the necessary new nomenclatural combinations are provided. The phylogenetic framework shows Phylloctenium, which is endemic to Madagascar and restricted to dry ecosystems, as basal and sister to the rest of the tribe, suggesting Madagascar to be the centre of origin of this clade. The remaining genera are diversified mostly in humid ecosystems, with evidence of multiple dispersals to the neighboring islands, including at least two to the Comoros, one to Mauritius and one to the Seychelles. Finally, we hypothesize that the ecological success of this tribe might have been triggered by a shift of fruit-dispersal mode from wind to lemur., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The origin of Neotropical biodiversity represents a key question in evolutionary biology. Despite the attempts to decipher the role of ecological and historical factors to present-day distribution patterns, robust phylogenetic studies of Neotropical clades are still needed before a comprehensive picture of the origin of Neotropical biodiversity can be achieved. Tynanthus Miers (Bignonieae, Bignoniaceae) is a well-circumscribed genus of Neotropical lianas with species that are narrowly distributed, except from a few taxa. The genus is characterized by a clove odor, small bilabiate flowers with the two upper lobes almost fused, and fruits with raised margins, all of which represent morphological synapomorphies for this clade. Other distinctive characters are the thecae reflexed forward, the densely pubescent ovaries and the poorly-developed nectariferous disk. The circumscription of the genus has remained constant over the years, despite the problematic limits of most genera of tribe Bignonieae. In this study, we reconstruct the phylogeny of Tynanthus based on two plastid (ndhF and rpl32-trnL) and one nuclear (pepC) markers and use this phylogenetic framework to investigate the biogeographical history of the genus. Our phylogenetic hypothesis provides further support for the monophyly of Tynanthus, and strongly supports a series of infra-generic clades. Most species are reconstructed as monophyletic, while T. cognatus and T. polyanthus are paraphyletic. Biogeographic reconstructions suggest that Tynanthus originated between 9.4 and 21.5Mya, most likely at approximately 15.3Mya. The MRCA of the genus was likely broadly distributed through lowland Amazonia, Western South America and Central America and diversified in the Neotropics during the Miocene. Closely related species are generally distributed within the same biogeographic area, suggesting that niche conservatism has played an important role in the diversification history of the group., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The ecological restoration strategies for highly threatened ecosystems such as the tropical dry forest, depend on the knowledge of limiting factors of biological processes for the different species. Some of these include aspects such as germination and seed longevity of typical species present in those forests. In this study, we evaluated the effect of light and temperature on seed germination of two Fabaceae (Samanea saman and Jacaranda caucana) and two Bignoniaceae (Pithecellobium dulce and Tabebuia rosea) species having potential use in restoration, and we analyzed the seed storage behavior of these species for a three months period. To study the light effect, four levels of light quality on seeds were used (photoperiod of 12 hours of white light, darkness and light enriched in red and far-red, both for an hour each day), and we combined them with three levels of alternated temperatures (20/25, 20/30 and 25/30*C-16/8h). For the storage behavior, two levels of seed moisture content particular for each species were used (low: 3.5-6.1% and high: 8.3-13.8%), with three storage temperatures (20, 5 and -20 degrees C) and two storage times (one and three months). The criterion for germination was radicle emergence which was measured in four replicates per treatment, and was expressed as percentage of germination (PG). There were significant differences in germination of Samanea saman and Jacaranda caucana among light and temperature treatments, with the lowest value in darkness treatments, whereas germination of Pithecellobium dulce and Tabebuia rosea did not differ between treatments (PG>90%). The most suitable temperature regime to promote germination in all species was 25/30 degrees C. These four species showed an orthodox seed storage behavior. We concluded that seeds of R dulce, J. caucana and T. rosea did not have an apparent influence of all light conditions tested in their germination response, which might confer advantages in colonization and establishment processes, while S. saman did not germinate well in darkness. We suggest the use of seeds of P dulce, J. caucana and T rosea in ecological restoration processes, due to their tolerance and germination under a wide range of temperature and light conditions. Futhermore, seeds of S. saman might be used in open areas such as forest gaps.

Background: Glaciations were recurrent throughout the Quaternary and potentially shaped species genetic structure worldwide by affecting population dynamics. Here, we implemented a multi-model inference approach to recover the distribution dynamics and demographic history of a Neotropical savanna tree, Tabebuia aurea (Bignoniaceae). Exploring different algorithms and paleoclimatic simulations, we used ecological niche modelling to generate alternative hypotheses of potential demographic changes through the last glacial cycle and estimated genetic parameters using coalescent modelling., Results: Comparing predictions from demographic hypotheses with genetic parameters of modern populations, our findings revealed a likely scenario of population decline, with spatial displacement towards Northeast Brazil from the last glacial maximum to the mid-Holocene. Subsequently, populations expanded in response to the return of the climatically suitable conditions in Central-West Brazil. Nevertheless, a wide historical refugium across Central Brazil likely maintained large populations connected throughout time. The expected genetic signatures from such predicted distribution dynamics are also corroborated by spatial genetic structure observed in modern populations., Conclusion: By exploring uncertainties inherent in multiple working hypotheses, we have shown that multi-model inference is a fruitful and efficient approach to recover the nature, timing and geographical context of the Tabebuia aurea population dynamic in response to the Quaternary climate changes.

The influence of ecological traits to the distribution and abundance of species is a prevalent issue in biodiversity science. Most studies of plant community assembly have focused on traits related to abiotic aspects or direct interactions among plants, with less attention paid to ignore indirect interactions, as those mediated by pollinators. Here, we assessed the influence of phylogeny, habitat, and floral morphology on ecological community structure in a clade of Neotropical lianas (tribe Bignonieae, Bignoniaceae). Our investigation was guided by the long-standing hypothesis that habitat specialization has promoted speciation in Bignonieae, while competition for shared pollinators influences species co-occurrence within communities. We analyzed a geo-referenced database for 94 local communities occurring across the Neotropics. The effect of floral morphological traits and abiotic variables on species co-occurrence was investigated, taking into account phylogenetic relationships. Habitat filtering seems to be the main process driving community assembly in Bignonieae, with environmental conditions limiting species distributions. Differing specialization to abiotic conditions might have evolved recently, in contrast to the general pattern of phylogenetic clustering found in communities of other diverse regions. We find no evidence that competition for pollinators affects species co-occurrence; instead, pollinator occurrence seems to have acted as an "environmental filter" in some habitats.

Birds play crucial role on the pollination of many plants. However, little is known about the interactions between nectarivorous neotropical birds and exotic Angiosperms. S. nilotica is an exotic African plant widely used in Brazilian urban landscaping. However, it has been poorly studied in relation to its interactions with Neotropical birds. In this way, we studied the feeding nectar strategies and the interspecific antagonistic behaviours among nectarivorous Neotropical birds to verify the bird contributions to the S. nilotica pollination. The study was conducted from May 2008 to April 2011, but only in months of S. nilotica flowering (April to May). From 148 hours of sampling we identified 16 species feeding nectar on S. nilotica: 13 hummingbirds (Trochilidae), Aratinga aurea (Psittacidae), Tangara palmarum (Thraupidae) and Coereba flaveola (Coerebidae). Eupetomena macroura was the most frequent (96.88%), followed by Chlorostilbon lucidus (78.13%) and Coereba flaveola (59.38%). Most birds obtained nectar by punching at the base of the corolla, except for A. aurea that obtained the nectar by the upper opening of the petals in 100% of its visits, Heliomaster furcifer (95.65%), F. fusca (95%) and A. nigricollis (70.27%). Despite E. macroura also obtains nectar only by punching at the base of the corolla, it showed the highest level of legitimate visits. Antagonistic events were more frequent in E. macroura (58.65%), Florisuga fusca (11.04%) and Amazilia fimbriata (10.87%), being E. macroura dominant in all events. These results showed E. macroura plays an important role on this plant being the most important bird as a potential pollinator. Moreover, other birds contribute partially to the S. nilotica pollination. Most probably it is a result of recent Neotropical bird interactions with this African plant.

Phenotypic integration is essential to the understanding of organismal evolution as a whole. In this study, a phylogenetic framework is used to assess phenotypic integration among the floral parts of a group of Neotropical lianas. Flowers consist of plant reproductive organs (carpels and stamens), usually surrounded by attractive whorls (petals and sepals). Thus, flower parts might be involved in different functions and developmental constraints, leading to conflicting selective forces. We found that Bignonieae flowers have very similar patterns of variance/covariance among traits and that such patterns are uncorrelated with the phylogenetic relationships between species. However, in spite of pattern stasis, our results also indicate that diversification of floral morphology in this group has occurred throughout the evolution of magnitudes of correlation among traits. Thus, we suggest that stabilizing selection has played an important role in phenotypic integration, resulting in the long-term stasis of covariance patterns underlying flower diversification during the ca. 50 Myr of evolution of Bignonieae. This is the first report of long-term stasis in the phenotypic integration of angiosperms, suggesting that patterns of floral morphology can be recognizable as specific attributes of distinct botanical families., (© 2013 The Authors. Journal of Evolutionary Biology © 2013 European Society For Evolutionary Biology.)

Much effort has been devoted to understanding the function of extrafloral nectaries (EFNs) for ant-plant-herbivore interactions. However, the pattern of evolution of such structures throughout the history of plant lineages remains unexplored. In this study, we used empirical knowledge on plant defences mediated by ants as a theoretical framework to test specific hypotheses about the adaptive role of EFNs during plant evolution. Emphasis was given to different processes (neutral or adaptive) and factors (habitat change and trade-offs with new trichomes) that may have affected the evolution of ant-plant associations. We measured seven EFN quantitative traits in all 105 species included in a well-supported phylogeny of the tribe Bignonieae (Bignoniaceae) and collected field data on ant-EFN interactions in 32 species. We identified a positive association between ant visitation (a surrogate of ant guarding) and the abundance of EFNs in vegetative plant parts and rejected the hypothesis of phylogenetic conservatism of EFNs, with most traits presenting K-values < 1. Modelling the evolution of EFN traits using maximum likelihood approaches further suggested adaptive evolution, with static-optimum models showing a better fit than purely drift models. In addition, the abundance of EFNs was associated with habitat shifts (with a decrease in the abundance of EFNs from forest to savannas), and a potential trade-off was detected between the abundance of EFNs and estipitate glandular trichomes (i.e. trichomes with sticky secretion). These evolutionary associations suggest divergent selection between species as well as explains K-values < 1. Experimental studies with multiple lineages of forest and savanna taxa may improve our understanding of the role of nectaries in plants. Overall, our results suggest that the evolution of EFNs was likely associated with the adaptive process which probably played an important role in the diversification of this plant group., (© 2012 The Authors. Journal of Evolutionary Biology © 2012 European Society For Evolutionary Biology.)

Aims: To evaluate the antiviral activity of Bignoniaceae species occurring in the state of Minas Gerais, Brazil., Methods and Results: Ethanol extracts of different anatomical parts of bignoniaceous plant species have been evaluated in vitro against human herpesvirus type 1 (HSV-1), vaccinia virus (VACV) and murine encephalomyocarditis virus (EMCV) by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A total of 34 extracts from 18 plant species selected according to ethnopharmacological and taxonomic criteria were screened. Fifteen of the 34 extracts (44.1%) have disclosed antiviral activity against one or more of the viruses assayed with EC(50) values in the range of 23.2 ± 2.5-422.7 ± 10.9 μg ml(-1)., Conclusions: Twelve of the 34 extracts (35.3%) might be considered promising sources of antiviral natural products, as they have shown EC50 ≤ 100 μg ml(-1). The present screening discloses the high potential of the Bignoniaceae family as source of antiviral agents., Significance and Impact of the Study: Active extracts were identified and deserve bioguided studies for the isolation of antiviral compounds and studies on mechanism of action., (© 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.)

Traditional medicine provides strong guidance for scientific experiments involving plant products used by the Brazilian people. The species "cipó-cravo" (Tynnanthus fasciculatus) is a plant commonly used either to combat indigestion and stomach aches, or as a general stimulant and aphrodisiac. In this study, the effects of "cipó-cravo" infusion were investigated within the testicular parenchyma of adult Wistar rats. Rats were divided into 3 groups: a control (distilled water) and two treated groups, which received the plant infusion (100 and 200mg/animal/day). The 200mg dose promoted a significant increase of the testicular parenchyma weight and of the volume and total length of the seminiferous tubules, as well as in total daily sperm production and sperm production per gram of testis.

For over a century, Catuaba has been used in Brazilian folk medicine as an aphrodisiac even though the identity of the plant material employed is often uncertain. The species recommended by the Brazilian Pharmacopeia is Anemopaegma arvense (Bignoniaceae), but many other plants, regionally known as Catuaba, are commercialised. Frequently, the quality control of such a complex system is based on chemical markers that do not supply a general idea of the system. With the advent of the metabolomics approach, a global analysis of samples becomes possible. It appears that (1)H-NMR is the most useful method for such application, since it can be used as a wide-spectrum chemical analysis technique. Unfortunately, the generated spectra is complex so a possible approach is to look at the metabolite profile as a whole using multivariate methods, for example, by application of principal component analysis (PCA). In the present paper, we describe for the first time a proton high-resolution magic angle spinning nuclear magnetic resonance ((1)H-HR-MAS NMR) method coupled with PCA for the metabolomic analysis of some commercial Catuaba samples, which provided a reduction in the time required for such analysis. A comparative study of HPLC, HR-MAS and liquid-NMR techniques is also reported.

Of the roughly 12,000 known plant species in Madagascar only 3% are found in the IUCN (World Conservation Union) Red List of Threatened Species. We assigned preliminary IUCN categories of threat to the species of a comparatively well-known tribe, Coleeae (Bignoniaceae), which comprises an endemic, species-rich radiation in Madagascar Because the IUCN Red List Categories and Criteria 3.1 discourage the use of the data-deficient category, we developed a novel method for differentiating between range-limited species and poorly sampled species. We used the Missouri Botanical Garden (MBG) gazetteer to determine where other collection efforts had taken place. We drew buffers around each Coleeae locality and determined how many times the surrounding area bad been visited since the last sighting of the specimens by intersecting the buffers with all known botanical localities from the MBG gazetteer We determined that at least 54% of the Coleeae species are threatened with extinction. Assignments of species to this category were often due to predicted future decline within their current area of occupancy and their lack of inclusion within the protected-area network (only 42% of species are known to occur in protected areas). Three species were presumed extinct, and an additional 12 have not been seen in decades. Among the species threatened with extinction, we "rescued" six of them from the data-deficient category by considering both the sample dates and localities of places where they occurred in relation to additional collections that took place in the immediate area. Due to their recent discovery, 15 species remained in the data-deficient category If Coleeae is representative of the Malagasy flora, or at least of other endemic-radiated plant groups, then species loss in Madagascar may be even more extreme than is realized.

A lignan glycoside [(+)-cycloolivil 4'-O-beta-d-glucopyranoside], a phenolic glycoside [3,4-dimethoxyphenyl 1-O-beta-d-xylopyranosyl-(1-->6)-beta-d-glucopyranoside] and a iridoid glycoside (stereospermoside) were isolated from the leaves and branches of Stereospermum cylindricum, together with (+)-cycloolivil, (+)-cycloolivil 6-O-beta-d-glucopyranoside, (-)-olivil, (-)-olivil 4-O-beta-d-glucopyranoside, (-)-olivil 4'-O-beta-d-glucopyranoside, vanilloloside, decaffeoyl-verbascoside, isoverbascoside, 3,4,5-trimethoxyphenyl 1-O-beta-d-xylopyranosyl-(1-->6)-beta-d-glucopyranoside, ajugol, verminoside, and specioside. The structure elucidations were based on spectroscopic evidence.