Unravelling the genetic differentiation among varieties of the Neotropical savanna tree Hancornia speciosa Gomes.

Background and Aims: Spatial distribution of species genetic diversity is often driven by geographical distance (isolation by distance) or environmental conditions (isolation by environment), especially under climate change scenarios such as Quaternary glaciations. Here, we used coalescent analyses coupled with ecological niche modelling (ENM), spatially explicit quantile regression analyses and the multiple matrix regression with randomization (MMRR) approach to unravel the patterns of genetic differentiation in the widely distributed Neotropical savanna tree, Hancornia speciosa (Apocynaceae). Due to its high morphological differentiation, the species was originally classified into six botanical varieties by Monachino, and has recently been recognized as only two varieties by Flora do Brasil 2020. Thus, H. speciosa is a good biological model for learning about evolution of phenotypic plasticity under genetic and ecological effects, and predicting their responses to changing environmental conditions. Methods: We sampled 28 populations (777 individuals) of Monachino's four varieties of H. speciosa and used seven microsatellite loci to genotype them. Key Results: Bayesian clustering showed five distinct genetic groups (K = 5) with high admixture among Monachino's varieties, mainly among populations in the central area of the species geographical range. Genetic differentiation among Monachino's varieties was lower than the genetic differentiation among populations within varieties, with higher within-population inbreeding. A high historical connectivity among populations of the central Cerrado shown by coalescent analyses may explain the high admixture among varieties. In addition, areas of higher climatic suitability also presented higher genetic diversity in such a way that the wide historical refugium across central Brazil might have promoted the long-term connectivity among populations. Yet, FST was significantly related to geographic distances, but not to environmental distances, and coalescent analyses and ENM predicted a demographical scenario of quasi-stability through time. Conclusions: Our findings show that demographical history and isolation by distance, but not isolation by environment, drove genetic differentiation of populations. Finally, the genetic clusters do not support the two recently recognized botanical varieties of H. speciosa, but partially support Monachino's classification at least for the four sampled varieties, similar to morphological variation.

Floral contrivances and specialised pollination mechanism strongly influence mixed mating in Wrightia tomentosa (Apocynaceae).

Reproductive success of a plant species is largely influenced by the outcome of mating pattern in a population. It is believed that a significantly larger proportion of animal-pollinated plants have evolved a mixed-mating strategy, the extent of which may vary among species. It is thus pertinent to investigate the key contributors to mating success, especially to identify the reproductive constraints in depauperate populations of threatened plant species. We examined the contribution of floral architecture, pollination mechanism and breeding system on the extent of outcrossing rate in a near-threatened tree species, Wrightia tomentosa. The breeding system was ascertained from controlled pollination experiments. In order to determine outcrossing rate, 60 open-pollinated progeny were analysed using an AFLP markers. Although the trees are self-compatible, herkogamy and compartmentalisation of pollen and nectar in different chambers of the floral tube effectively prevent spontaneous autogamy. Pollination is achieved through specialised interaction with moths. Differential foraging behaviour of settling moths and hawkmoths leads to different proportions of geitonogamous and xenogamous pollen on the stigma. However, most open-pollinated progeny were the result of xenogamy (outcrossing rate, tm = 0.68). The study shows that floral contrivances and pollination system have a strong influence on mating pattern. The differential foraging behaviour of the pollinators causes deposition of a mixture of self- and cross-pollen to produce a mixed brood. Inbreeding depression and geitonogamy appear to play a significant role in sustaining mixed mating in this species.

A novel function for globulin in sequestering plant hormone: Crystal structure of Wrightia tinctoria 11S globulin in complex with auxin.

Auxin levels are tightly regulated within the plant cell, and its storage in the isolated cavity of proteins is a measure adopted by cells to maintain the availability of auxin. We report the first crystal structure of Wrightia tinctoria 11S globulin (WTG) in complex with Indole-3-acetic acid (IAA), an auxin, at 1.7 Å resolution. WTG hexamers assemble as a result of the stacking interaction between the hydrophobic surfaces of two trimers, leaving space for the binding of charged ligands. The bound auxin is stabilized by non-covalent interactions, contributed by four chains in each cavity. The presence of bound ligand was confirmed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and high-resolution mass spectrometry (HRMS). Here, we hypothesize that the cleavage of globulins by endopeptidases leads to the movement of the hydrophilic loop region from the surface to the periphery, leaving space for the binding of auxin, and promotes hexamer formation. As the process of germination proceeds, there is a change in the pH, which induces the dissociation of the hexamer and the release of auxin. The compact hexameric assembly ensures the long-term, stable storage of the hormone. This suggests a role for globulin as a novel player in auxin homeostasis.

Pollinia transfer on moth legs in Hoya carnosa (Apocynaceae).

PREMISE OF THE STUDY: The genus Hoya (Apocynaceae; Asclepiadoideae) is characterized by a set of complex floral characters unique among the asclepiads, but their role in pollination is poorly understood. Here, we report a new mechanism of asclepiad pollination in the wax plant Hoya carnosa: the pollinaria are transferred on the legs of medium or large settling moths. METHODS: Floral visitors and their behavior were observed on Amami-Oshima Island during 2013-2015, and the efficacy of different pollinators was determined by counting the pollinarium loads on different flower visitors. The floral characters were studied to establish the process of pollination in relation to pollinator behavior on the flower. KEY RESULTS: Hoya carnosa was visited by various settling moths at night, but pollinia attachment was confirmed predominantly on the legs of the large moth Erebus ephesperis (Noctuidae) and less frequently on the legs of the medium-sized moths Bastilla arcuata (Crambidae) and Cleora injectaria (Geometridae). The moths walked around and searched for nectar on the inflorescence, and the corpusculum became clipped to the arolia (adhesive pads on moth tarsi) as they stepped on the pollinaria between the staminal corona. The downward spherical inflorescence of aggregated flowers with flat, velvety petals and a slippery corona provides restricted footholds for the visitors, which efficiently leads pollinator legs to the pollinaria. CONCLUSIONS: Although the pollination system of Hoya is largely unknown, pollination by insect legs may be a major pollination system in this genus because these basic floral characters are shared among many species.

Ceropegia sandersonii Mimics Attacked Honeybees to Attract Kleptoparasitic Flies for Pollination.

Four to six percent of plants, distributed over different angiosperm families, entice pollinators by deception [1]. In these systems, chemical mimicry is often used as an efficient way to exploit the olfactory preferences of animals for the purpose of attracting them as pollinators [2,3]. Here, we report a very specific type of chemical mimicry of a food source. Ceropegia sandersonii (Apocynaceae), a deceptive South African plant with pitfall flowers, mimics attacked honeybees. We identified kleptoparasitic Desmometopa flies (Milichiidae) as the main pollinators of C. sandersonii. These flies are well known to feed on honeybees that are eaten by spiders, which we thus predicted as the model chemically mimicked by the plant. Indeed, we found that the floral scent of C. sandersonii is comparable to volatiles released from honeybees when under simulated attack. Moreover, many of these shared compounds elicited physiological responses in antennae of pollinating Desmometopa flies. A mixture of four compounds-geraniol, 2-heptanone, 2-nonanol, and (E)-2-octen-1-yl acetate-was highly attractive to the flies. We conclude that C. sandersonii is specialized on kleptoparasitic fly pollinators by deploying volatiles linked to the flies' food source, i.e., attacked and/or freshly killed honeybees. The blend of compounds emitted by C. sandersonii is unusual among flowering plants and lures kleptoparasitic flies into the trap flowers. This study describes a new example of how a plant can achieve pollination through chemical mimicry of the food sources of adult carnivorous animals.

Long-term persisting hybrid swarm and geographic difference in hybridization pattern: genetic consequences of secondary contact between two Vincetoxicum species (Apocynaceae-Asclepiadoideae).

BACKGROUND: During glacial periods, glacial advances caused temperate plant extirpation or retreat into localized warmer areas, and subsequent postglacial glacial retreats resulted in range expansions, which facilitated secondary contact of previously allopatric isolated lineages. The evolutionary outcomes of secondary contact, including hybrid zones, dynamic hybrid swarm, and resultant hybrid speciation, depends on the strengths of reproductive barriers that have arisen through epistatic and pleiotropic effects during allopatric isolation. The aim of this study was to demonstrate refugia isolation and subsequent secondary contact between two perennial Asclepioid species and to assess the genetic consequences of the secondary contact. We modeled the range shift of two ecologically distinct Vincetoxicum species using the species distribution model (SDM) and assessed the genetic consequences of secondary contact by combining morphological and genetic approaches. We performed morphometric analysis (592 individuals) and examined 10 nuclear microsatellites (671 individuals) in V. atratum, V. japonicum, and putative hybrid populations. RESULTS: Multivariate analysis, model-based Bayesian analysis, and non-model-based discriminant analysis of principal components confirmed the hybridization between V. atratum and V. japonicum. High pollen fertility and a lack of linkage disequilibrium suggested that the hybrid populations may be self-sustaining and have persisted since V. atratum and V. japonicum came into contact during the post-glacial period. Moreover, our findings show that the pattern of hybridization between V. atratum and V. japonicum is unidirectional and differs among populations. Geographically-isolated hybrid populations exist as genetically distinct hybrid swarms that consist of V. atratum-like genotypes, V. japonicum-like genotypes, or admixed genotypes. In addition, Bayesian-based clustering analysis and coalescent-based estimates of long-term gene flow showed patterns of introgressive hybridization in three morphologically 'pure' V. japonicum populations. CONCLUSION: In this study, we demonstrated that climatic oscillations during the Quaternary period likely led to species range shift and subsequently secondary contact. Hybrid populations may be self-sustaining and have persisted since V. atratum and V. japonicum came into contact during the post-glacial period. Pattern of hybridization between V. atratum and V. japonicum is unidirectional and differs among populations. We concluded that these differences in the genetic consequences of secondary contact are caused by historical colonization processes and/or natural selection.

Use of spatial analysis to test hypotheses on plant recruitment in a hyper-arid ecosystem.

Mounds originating from wind-blown sediment accumulation beneath vegetation (nebkhas) often indicate land degradation in dry areas. Thus far, most nebkha research has focused on individual plants. Here, we aimed to explore population-scale processes (up to scales of about 100 m) that might explain an observed nebkha landscape pattern. We mapped the Rhazya stricta Decne. population in a 3 ha study site in a hyper-arid region of Saudi Arabia. We compared the spatial patterns of five different cohorts (age classes) of observed nebkha host plants to those expected under several hypothesized drivers of recruitment and intraspecific interaction. We found that all R. stricta cohorts had a limited fractional vegetation cover and established in large-scale clusters. This clustering weakened with cohort age, possibly indicating merging of neighboring vegetation patches. Different cohort clusters did not spatially overlap in most cases, indicating that recruitment patterns changed position over time. Strong indications were found that the main drivers underlying R. stricta spatial configurations were allogenic (i.e. not driven by vegetation) and dynamic. Most likely these drivers were aeolian-driven sand movement or human disturbance which forced offspring recruitment in spatially dynamic clusters. Competition and facilitation were likely active on the field site too, but apparently had a limited effect on the overall landscape structure.

High specialisation in the pollination system of Mandevilla tenuifolia (J.C. Mikan) Woodson (Apocynaceae) drives the effectiveness of butterflies as pollinators.

Butterfly pollination in the tropics is considered somewhat effective or solely effective in a few plant species. In the present study, we tested the hypothesis that Mandevilla tenuifolia (Apocynaceae), which has floral attributes associated with psychophily, has strategies adapted to pollination by butterflies, restricting other floral visitors and making these insects act as efficient pollinators. We analysed the floral and reproductive biology of M. tenuifolia, as well as the frequency and efficiency of its flower visitors. M. tenuifolia is an herb whose flowers have strong herkogamy and secondary pollen presentation on the style head, which corresponds to 60.4% of pollen on the anthers. Flower longevity and the long period of receptivity of the stigmatic region associated with the large amount of pollen removed in the first visits suggest that flowers remain functionally female during part of anthesis. Butterflies, mainly of the families Nymphalidae and Pieridae, are the only pollinators of M. tenuifolia. Despite being self-compatible, M. tenuifolia depends on biotic vectors for fruit production. A non-significant difference in fruit set between controlled treatments and natural conditions suggests that the pollinators are efficient. The inclination resulting from the landing of butterflies on flowers, together with flower morphology, guiding the insect proboscis inside the floral tube, as well as the frequency and efficiency of butterfly visits, are evidence of the close relationship between butterflies and M. tenuifolia, and also of the efficiency of these insects as pollinators.

The search for Pleiades in trait constellations: functional integration and phenotypic selection in the complex flowers of Morrenia brachystephana (Apocynaceae).

Pollinator-mediated natural selection on single traits, such as corolla tube or spur length, has been well documented. However, flower phenotypes are usually complex, and selection is expected to act on several traits that functionally interact rather than on a single isolated trait. Despite the fact that selection on complex phenotypes is expectedly widespread, multivariate selection modelling on such phenotypes still remains under-explored in plants. Species of the subfamily Asclepiadoideae (Apocynaceae) provide an opportunity to study such complex flower contrivances integrated by fine-scaled organs from disparate developmental origin. We studied the correlation structure among linear floral traits (i) by testing a priori morphological, functional or developmental hypotheses among traits and (ii) by exploring the organization of flower covariation, considering alternative expectations of modular organization or whole flower integration through conditional dependence analysis (CDA) and integration matrices. The phenotypic selection approach was applied to determine whether floral traits involved in the functioning of the pollination mechanism were affected by natural selection. Floral integration was low, suggesting that flowers are organized in more than just one correlation pleiad; our hypothetical functional correlation matrix was significantly correlated with the empirical matrix, and the CDA revealed three putative modules. Analyses of phenotypic selection showed significant linear and correlational gradients, lending support to expectations of functional interactions between floral traits. Significant correlational selection gradients found involved traits of different floral whorls, providing evidence for the existence of functional integration across developmental domains.

C3 photosynthesis in the desert plant Rhazya stricta is fully functional at high temperatures and light intensities.

The C3 plant Rhazya stricta is native to arid desert environment zones, where it experiences daily extremes of heat, light intensity (PAR) and high vapour pressure deficit (VPD). We measured the photosynthetic parameters in R. stricta in its native environment to assess the mechanisms that permit it to survive in these extreme conditions. Infrared gas exchange analysis examined diel changes in assimilation (A), stomatal conductance (gs ) and transpiration (E) on mature leaves of R. stricta. A/ci analysis was used to determine the effect of temperature on carboxylation capacity (Vc,max ) and the light- and CO2 -saturated rate of photosynthesis (Amax ). Combined chlorophyll fluorescence and gas exchange light response curve analysis at ambient and low oxygen showed that both carboxylation and oxygenation of Rubisco acted as the major sinks for the end products of electron transport. Physiological analysis in conjunction with gene expression analysis suggested that there are two isoforms of Rubisco activase which may provide an explanation for the ability of R. stricta to maintain Rubisco function at high temperatures. The potential to exploit this ability to cope with extreme temperatures is discussed in the context of future crop improvement.

When bigger is not better: intraspecific competition for pollination increases with population size in invasive milkweeds.

One of the essential requirements for an introduced plant species to become invasive is an ability to reproduce outside the native range, particularly when initial populations are small. If a reproductive Allee effect is operating, plants in small populations will have reduced reproductive success relative to plants in larger populations. Alternatively, if plants in small populations experience less competition for pollination than those in large populations, they may actually have higher levels of reproductive success than plants in large populations. To resolve this uncertainty, we investigated how the per capita fecundity of plants was affected by population size in three invasive milkweed species. Field surveys of seed production in natural populations of different sizes but similar densities were conducted for three pollinator-dependent invasive species, namely Asclepias curassavica, Gomphocarpus fruticosus and G. physocarpus. Additionally, supplemental hand-pollinations were performed in small and large populations in order to determine whether reproductive output was limited by pollinator activity in these populations. Reproductive Allee effects were not detected in any of the study species. Instead, plants in small populations exhibited remarkably high levels of reproductive output compared to those in large populations. Increased fruit production following supplemental hand-pollinations suggested that the lower reproductive output of naturally pollinated plants in large populations is a consequence of pollen limitation rather than limitation due to abiotic resources. This is consistent with increased intraspecific competition for pollination amongst plants in large populations. It is likely that the invasion of these milkweed species in Australia has been enhanced because plants in small founding populations experience less intraspecific competition for pollinators than those in large populations, and thus have the ability to produce copious amounts of seeds.

Generalised pollination systems for three invasive milkweeds in Australia.

Because most plants require pollinator visits for seed production, the ability of an introduced plant species to establish pollinator relationships in a new ecosystem may have a central role in determining its success or failure as an invader. We investigated the pollination ecology of three milkweed species - Asclepias curassavica, Gomphocarpus fruticosus and G. physocarpus - in their invaded range in southeast Queensland, Australia. The complex floral morphology of milkweeds has often been interpreted as a general trend towards specialised pollination requirements. Based on this interpretation, invasion by milkweeds contradicts the expectation than plant species with specialised pollination systems are less likely to become invasive that those with more generalised pollination requirements. However, observations of flower visitors in natural populations of the three study species revealed that their pollination systems are essentially specialised at the taxonomic level of the order, but generalised at the species level. Specifically, pollinators of the two Gomphocarpus species included various species of Hymenoptera (particularly vespid wasps), while pollinators of A. curassavica were primarily Lepidoptera (particularly nymphalid butterflies). Pollinators of all three species are rewarded with copious amounts of highly concentrated nectar. It is likely that successful invasion by these three milkweed species is attributable, at least in part, to their generalised pollinator requirements. The results of this study are discussed in terms of how data from the native range may be useful in predicting pollination success of species in a new environment.

Differential pollinator effectiveness and importance in a milkweed (Asclepias, Apocynaceae) hybrid zone.

PREMISE OF THE STUDY: Exceptions to the ideal of complete reproductive isolation between species are commonly encountered in diverse plant, animal, and fungal groups, but often the causative ecological processes are poorly understood. In flowering plants, the outcome of hybridization depends in part on the effectiveness of pollinators in interspecific pollen transport. In the Asclepias exaltata and A. syriaca (Apocynaceae) hybrid zone in Shenandoah National Park, Virginia, extensive introgression has been documented. The objectives of this study were to (1) determine the extent of pollinator overlap among A. exaltata, A. syriaca, and their hybrids and (2) identify the insect taxa responsible for hybridization and introgression. METHODS: We observed focal plants of parental species and hybrids to measure visitation rate, visit duration, and per-visit pollinia removal and deposition, and we calculated pollinator effectiveness and importance. KEY RESULTS: Visitation rates varied significantly between the 2 yr of the study. Overall, Apis mellifera, Bombus sp., and Epargyreus clarus were the most important pollinators. However, Bombus sp. was the only visitor that was observed to both remove and insert pollinia for both parent species as well as hybrids. CONCLUSIONS: We conclude that Bombus may be a key agent of hybridization and introgression in these sympatric milkweed populations, and hybrids are neither preferred nor selected against by pollinators. Thus, we have identified a potential mechanism for how hybrids act as bridges to gene flow between A. exaltata and A. syriaca. These results provide insights into the breakdown of prezygotic isolating mechanisms.

Functional morphology and wasp pollination of two South American asclepiads (Asclepiadoideae-Apocynaceae).

UNLABELLED: BACKGROUND AND AIMS The extreme complexity of asclepiad flowers (Asclepiadoideae-Apocynaceae) has generated particular interest in the pollination biology of this group of plants especially in the mechanisms involved in the pollination processes. This study compares two South American species, Morrenia odorata and Morrenia brachystephana, with respect to morphology and anatomy of flower structures, dynamic aspects of the pollination mechanism, diversity of visitors and effectiveness of pollinators. METHODS: Floral structure was studied with fresh and fixed flowers following classical techniques. The pollination mechanism was studied by visiting fresh flowers in the laboratory with artificial pollinator body parts created with an eyelash. Morphometric and nectar measurements were also taken. Pollen transfer efficiency in the flowers was calculated by recording the frequency of removed and inserted pollinia. Visitor activity was recorded in the field, and floral visitors were captured for subsequent analysis of pollen loads. Finally, pollinator effectiveness was calculated with an index. KEY RESULTS: The detailed structure of the flowers revealed a complex system of guide rails and chambers precisely arranged in order to achieve effective pollinaria transport. Morrenia odorata is functionally specialized for wasp pollination, and M. brachystephana for wasp and bee pollination. Pollinators transport chains of pollinaria adhered to their mouthparts. CONCLUSIONS: Morrenia odorata and M. brachystephana present differences in the morphology and size of their corona, gynostegium and pollinaria, which explain the differences in details of the functioning of the general pollination mechanism. Pollination is performed by different groups of highly effective pollinators. Morrenia species are specialized for pollination mainly by several species of wasps, a specialized pollination which has been poorly studied. In particular, pompilid wasps are reported as important pollinators in other regions outside South Africa. A putative new function of nectar in asclepiads is presented, as it would be contributing to the pollination mechanism.

The role of inbreeding and outbreeding in herbivore resistance and tolerance in Vincetoxicum hirundinaria.

BACKGROUND AND AIMS: Inbreeding via self-fertilization may have negative effects on plant fitness (i.e. inbreeding depression). Outbreeding, or cross-fertilization between genetically dissimilar parental plants, may also disrupt local adaptation or allelic co-adaptation in the offspring and again lead to reduced plant fitness (i.e. outbreeding depression). Inbreeding and outbreeding may also increase plant vulnerability to natural enemies by altering plant quality or defence. The effects of inbreeding and outbreeding on plant size and response to herbivory in the perennial herb, Vincetoxicum hirundinaria, were investigated. METHODS: Greenhouse experiments were conducted using inbred and outbred (within- and between-population) offspring of 20 maternal plants from four different populations, quantifying plant germination, size, resistance against the specialist folivore, Abrostola asclepiadis, and tolerance of simulated defoliation. KEY RESULTS: Selfed plants were smaller and more susceptible to damage by A. asclepiadis than outcrossed plants. However, herbivore biomass on selfed and outcrossed plants did not differ. The effects of inbreeding on plant performance and resistance did not differ among plant populations or families, and no inbreeding depression at all was found in tolerance of defoliation. Between-population outcrossing had no effect on plant performance or resistance against A. asclepiadis, indicating a lack of outbreeding depression. CONCLUSIONS: Since inbreeding depression negatively affects plant size and herbivore resistance, inbreeding may modify the evolution of the interaction between V. hirundinaria and its specialist folivore. The results further suggest that herbivory may contribute to the maintenance of a mixed mating system of the host plants by selecting for outcrossing and reduced susceptibility to herbivore attack, and thus add to the growing body of evidence on the effects of inbreeding on the mating system evolution of the host plants and the dynamics of plant-herbivore interactions.

Morpho-anatomical characters of the leaf and stem of Mandevilla coccinea (Hook. et Arn.) Woodson, Apocynaceae / Características morfo-anatômicas de folha e caule de Mandevilla coccinea (Hook. et Arn.) Woodson, Apocynaceae

Mandevilla coccinea (Hook. et Arn.) Woodson, Apocynaceae is a herb native to South America employed in folk medicine as an analgesic, anti-inflammatory and to inhibit snake venom effects. This work was carried out to study the morpho-anatomical characters of the leaf and stem in order to advance knowledge on this medicinal plant and on pharmacognostic quality control. The plant material was fixed and prepared according to light and scanning electron microtechniques. Its leaves are simple, alternate and ovate-obovate. The epidermis is uniseriate and coated with a thick and striate cuticle. The stomata are paracytic and occur on both foliar surfaces. The dorsiventral-like mesophyll has a sub-epidermal parenchymatic layer containing phenolic substances and is traversed by minor collateral vascular bundles. The midrib is biconvex and the petiole is concave-convex, both presenting bicollateral vascular bundles in an open arc. The stem is circular in transverse section and the epidermis remains in incipient secondary growth. A sub-epidermal parenchymatic layer containing phenolic substances, a discontinuous sclerenchymatic sheath of non-lignified fibres and cylinders of external phloem, xylem and internal phloem occur. Numerous branched laticifers and idioblasts with phenolic substances are present in the leaf and stem.

Mandevilla coccinea (Hook. et Arn.) Woodson, Apocynaceae, é uma espécie herbácea nativa da América do Sul e empregada na medicina popular como analgésico, antiinflamatório e para inibir os efeitos de veneno de cobra. Este trabalho objetivou estudar os caracteres morfoanatômicos de folha e caule, a fim de fornecer conhecimento dessa espécie medicinal e para o controle de qualidade farmacognóstico. O material foi fixado e preparado de acordo com técnicas usuais de microscopia de luz e eletrônica de varredura. As folhas são simples, alternas e ovado-obovadas. A epiderme é unisseriada e recoberta por uma cutícula espessada e estriada. Os estômatos são paracíticos e ocorrem em ambas as superfícies foliares. O mesofilo tende a ser dorsiventral, apresenta uma camada subepidérmica parenquimática contendo compostos fenólicos e é percorrido por feixes vasculares colaterais de pequeno porte. A nervura central é biconvexa e o pecíolo é côncavo-convexo, ambos apresentando feixes vasculares bicolaterais em arco aberto no parênquima fundamental. O caule tem secção transversal circular e, em estrutura secundária incipiente, a epiderme permanece. Na sequência, encontram-se camada subepidérmica parenquimática contendo compostos fenólicos, bainha esclerenquimática descontínua composta de fibras não lignificadas, além de cilindros de floema externo, xilema e floema interno. Numerosos laticíferos ramificados e idioblastos com substâncias fenólicas estão presentes na folha e no caule.

Associations of plant fitness, leaf chemistry, and damage suggest selection mosaic in plant-herbivore interactions.

The geographic mosaic theory of coevolution states that variation in species interactions forms the raw material for coevolutionary processes, which take place over large geographic scales. One key assumption underlying the process of coevolution in plant-herbivore interactions is that herbivores exert selection on their host plants and that this selection varies among plant populations. We examined spatial variation in the existence and strength of phenotypic selection on host plant resistance exerted by specialist herbivores in 17 archipelago populations of the perennial herb Vincetoxicum hirundinaria (Asclepiadaceae). In these highly fragmented populations, V. hirundinaria is consumed by the larvae of two specialist herbivores: the folivorous moth Abrostola asclepiadis and the seed predator Euphranta connexa. Selection imposed on host plants by these herbivores was examined by analyzing the associations between levels of herbivory, plant fitness, and contents of a number of leaf chemicals reflecting plant resistance to and quality for the herbivores. We found extensive spatial variation in the levels of herbivory and in plant fitness. More importantly, the impact of both leaf herbivory and seed predation on plant fitness varied among plant populations, indicating spatial variation in phenotypic selection. In addition, leaf chemistry varied widely among plant populations, reflecting spatial variation in plant quality as food for the herbivores. However, leaf compounds influenced folivory similarly in all the studied plant populations, and interestingly, some of the compounds were associated with the intensity of seed predation. Finally, some of the leaf compounds were associated with plant fitness, and the strength and direction of these associations varied among plant populations. The observed spatial variation in the strength of the interactions between V. hirundinaria and its specialist herbivores suggests a geographic selection mosaic. Because the occurrence and strength of spatial variation varied between the two specialist herbivores, our results highlight the importance of considering multiple enemies when trying to understand evolution of interactions between plants and their herbivores.

Proteomic study of Carissa spinarum in response to combined heat and drought stress.

Carissa spinarum is one of the secondary advantage plants grown in dry-hot valleys in China, which can survive under stress conditions of high temperature and extreme low humidity. Here, we studied the physiological and proteomic changes of C. spinarum in response to 42 degrees C heat stress treatment in combination with drought stress. Dynamic changes in the leaf proteome were analyzed at four time points during the stress treatment and recovery stages. Approximately, 650 protein spots were reproducibly detected in each gel. Forty-nine spots changed their expression levels upon heat and drought treatment, and 30 proteins were identified by MS and 2-D Western blot. These proteins were classified into several categories including HSP, photosynthesis-related protein, RNA-processing protein and proteins involved in metabolism and energy production. The potential roles of these stress-responsive proteins are discussed.

Palp-faction: an African milkweed dismembers its wasp pollinators.

Interactions between pollinators and nectar-producing flowers are usually assumed to be mutualistic, but the exploitative basis of these relationships can lead to antagonistic interactions. Flowers of the African milkweed, Pachycarpus appendiculatus E. Mey, produce concentrated nectar that is consumed primarily by the large spider-hunting wasp Hemipepsis dedjas Guerin (Hymenoptera: Pompilidae). Pollinaria of this milkweed become attached to the palps of these wasps during nectar feeding. Broken wasp palps were found between guide rails, attached to corpuscula that were trapped behind the guide rails, and attached to pollinia that were inserted into the stigmatic chambers of the flowers. Approximately 85% of wasps captured on flowers of P. appendiculatus were missing one or more palps, whereas only 9% of wasps captured on flowers of another asclepiad species were missing any palps. It thus seems that wasps face a high risk of losing their palps when foraging on these flowers. The interaction may thus be antagonistic for the wasps if the cost of losing their sensory palps (not yet established) is greater than the benefits of the nectar reward. The plants, however, gain clear benefit from the interaction, as verified by the removal and insertion of pollinia in flowers exposed solely to visits by pompilid wasps.

Fly pollination in Ceropegia (Apocynaceae: Asclepiadoideae): biogeographic and phylogenetic perspectives.

BACKGROUND AND AIMS: Ceropegia (Apocynaceae subfamily Asclepiadoideae) is a large, Old World genus of >180 species, all of which possess distinctive flask-shaped flowers that temporarily trap pollinators. The taxonomic diversity of pollinators, biogeographic and phylogenetic patterns of pollinator exploitation, and the level of specificity of interactions were assessed in order to begin to understand the role of pollinators in promoting diversification within the genus. METHODS: Flower visitor and pollinator data for approx. 60 Ceropegia taxa were analysed with reference to the main centres of diversity of the genus and to a cpDNA-nrDNA molecular phylogeny of the genus. KEY RESULTS: Ceropegia spp. interact with flower-visiting Diptera from at least 26 genera in 20 families, of which 11 genera and 11 families are pollinators. Size range of flies was 0.5-4.0 mm and approx. 94 % were females. Ceropegia from particular regions do not use specific fly genera or families, though Arabian Peninsula species are pollinated by a wider range of Diptera families than those in other regions. The basal-most clade interacts with the highest diversity of Diptera families and genera, largely due to one hyper-generalist taxon, C. aristolochioides subsp. deflersiana. Species in the more-derived clades interact with a smaller diversity of Diptera. Approximately 60 % of taxa are so far recorded as interacting with only a single genus of pollinators, the remaining 40 % being less conservative in their interactions. Ceropegia spp. can therefore be ecological specialists or generalists. CONCLUSIONS: The genus Ceropegia has largely radiated without evolutionary shifts in pollinator functional specialization, maintaining its interactions with small Diptera. Intriguing biogeographic and phylogenetic patterns may reflect processes of regional dispersal, diversification and subsequent specialization onto a narrower range of pollinators, though some of the findings may be caused by inconsistent sampling. Comparisons are made with other plant genera in the Aristolochiaceae and Araceae that have evolved flask-shaped flowers that trap female flies seeking oviposition sites.