Hancornia speciosa Gomes Latex Increases Bone Mineralization in Rats: A Preclinical Study / Látex de Hancornia speciosa Gomes aumenta a mineralização óssea em ratos: Um estudo pré-clínico

Abstract Objective To evaluate the systemic effect of Hancornia speciosa latex on bone neoformation and mineralization in rats. Methods For that, the latex was first collected, and its composition was analyzed. A total of 30 male Wistar rats were used, which were simultaneously submitted to two surgical procedures: extraction of an incisor and creation of a defect with 2 mm in diameter in the parietal bone. The rats were divided into two groups: systemic control (SC) systemic latex (SX) which were administered, orally and daily, 1.5 mL of water or a solution containing 50% of water and 50% of latex by gavage, respectively. After 15 days of the treatment, the animals were euthanized and their samples were collected. Results The results were statistically analyzed, and the level of significance was set at 0.05. We showed that H. speciosa latex contained calcium. The oral and daily administration of the latex for 15 days increased the contents of calcium and phosphorus in the basal bone and newly-formed bone in the mandibular alveolus of rats. Conclusion The present was a pioneer study demonstrating the potential of H. speciosa latex in increasing bone mineralization. Our results may aid in the conception and development of a natural drug.

Resumo Objetivo Avaliar o efeito sistêmico do látex de Hancornia especiosa na neoformação óssea e mineralização em ratos. Métodos Para isso, primeiro o látex foi coletado, e sua composição foi analisada. No estudo, foram utilizados 30 ratos Wistar machos submetidos simultaneamente a dois procedimentos cirúrgicos: extração de incisivo e criação de um defeito de 2 mm de diâmetro no osso parietal. Os ratos foram divididos em dois grupos: controle sistêmico (CS) e látex sistêmico (XS), aos quais foi administrado, oral e diariamente, 1,5 mL de água ou uma solução contendo 50% de água e 50% de látex por gavagem, respectivamente. Após 15 dias do tratamento, os animais foram eutanizados, e suas amostras, coletadas. Resultados Os resultados foram analisados estatisticamente, e o nível de significância foi fixado em 0,05. Mostramos que o látex de H. speciosa continha cálcio. A administração oral e diária deste látex por 15 dias aumentou o conteúdo de cálcio e fósforo de osso basal e de osso recém-formado no alvéolo mandibular de ratos. Conclusão Este foi um estudo pioneiro, que demonstrou o potencial do látex de H. speciosa no aumento da mineralização óssea. Nossos resultados podem ajudar na concepção e no desenvolvimento de uma droga natural.

Morfo-anatomía foliar de Araujia odorata (Apocynaceae-Asclepiadoideae) en un gradiente altitudinal: histoquímica de laticíferos / Foliar morpho-anatomy of Araujia odorata (Apocynaceae-Asclepiadoideae) in an altitudinal gradient: histochemistry of laticifers

Araujia odorata is a sub-shrub native from Argentina, Brazil, Paraguay and Uruguay, whose latex, roots and leaves are used in traditional medicine. The objective of this work is to study the foliar morpho-anatomy of six populations in an altitudinal gradient (359-2155 m.a.s.l.) of Northwestern Argentine and to determine the nature of the compounds present in the laticiferous of the stems and fruits using conventional techniques for plant anatomy. The populations under study did not show significant morpho-anatomical differences. They presented simple leaves, pinnated brochydodromous venation, amphiestomatic isolateral lamina, brachy, anomo and amphicyclocytic stomata, eglandular trichomes, midvein with bicolateral vascular bundle and non-articulated laticifers continuous in the petiole, stem and fruits. Differences in the quantified foliar parameters are observed, however, only the density of trichomes, stomata and the thickness of the cuticle are positively correlated with the altitudinal gradient, indicating phenotypic plasticity. Histochemical analysis of laticifers and other stem idioblasts of A. odoratawas performed for the first time.

Araujia odorata, es un subarbusto nativo de Argentina, Brasil, Paraguay y Uruguay, cuyo látex, raíces y hojas son utilizados en medicina popular. Se plantea como objetivo realizar un estudio morfo-anatómico foliar de seis poblaciones del Noroeste Argentino en un gradiente altitudinal (359-2155 m.s.n.m) y determinar la naturaleza de los compuestos presentes en laticíferos de tallos y frutos mediante técnicas convencionales de anatomía vegetal. Las poblaciones estudiadas no evidenciaron diferencias morfo-anatómicas significativas. Presentan hojas simples, venación pinnada broquidódroma, lámina isolateral anfiestomática, estomas braqui, anomo y anficiclocíticos, tricomas eglandulares, nervio medio con haz bicolateral y laticíferos no-articulados continuos en pecíolo, tallo y frutos. Se observan diferencias en los parámetros foliares cuantificados, sin embargo, solo la de densidad de tricomas, estomas y el espesor de cutícula se correlacionan positivamente con el gradiente altitudinal indicando plasticidad fenotípica. Se realiza por primera vez un análisis histoquímico de los laticíferos y otros idioblastos del tallo A. odorata.

Foliar micromorphology and its role in identification of the Apocynaceae taxa.

In the present study, we evaluate the importance of foliar epidermal micromorphological characteristics of Apocyanaceae for accurate identification and classification. The species were collected from the University of Peshawar's main campus in the spring season to observe its qualitative and quantitative features. The length and width of guard cells, stomatal pore and subsidiary cells, trichomes, and crypts on both sides of the leaf were examined. Many species were observed to be hypostomatic. Plumeria rubra, Raulfia serpentine, Thevetia peruviana, Trachelospermum lucidum, Alstonia scholaris, and Catharanthus roseus demonstrated hypostomatic leaves. Nearly all the investigated species had anisocytic type of stomata only or in combination with other types of stomata on the upper and lower epidermis. Carissa carandas had anomocytic, anisocytic, and cyclocytic type of stomata on the upper epidermis, and the lower epidermis showed variations in stomatal type, such as anomocytic, stephanocytic, brachyparacytic, and hemiparacytic. Nerium oleander had no specific shape of stomata but showed stomatal crypts in which the stomata were enclosed inside many trichomes. The taxonomic key based on stomatal types, epidermal cells, stomatal index value, and statistical analysis, along with the variations in the epidermal cells, shows the link between the selected plants species, which will provide a baseline for future anatomical studies. This study highlights many undocumented micromorphological characteristics. The anatomical characteristics observed in this study will be helpful for taxonomic identification and species delimitation of the family Apocynaceae.

Molecular Networking Reveals Serpentinine-Related Bisindole Alkaloids from Picralima nitida, a Previously Well-Investigated Species.

Five new monoterpene indole alkaloids (1-5), including four serpentinine-related bisindoles and one alstonine derivative monomer, have been isolated from the aerial parts of Picralima nitida. Their structures were elucidated by analysis of their HRMS and NMR spectroscopic data, and their absolute configurations were deduced from the comparison of experimental and simulated ECD spectra. In addition, two known compounds (6 and 7), previously undescribed from P. nitida, have also been purified. The compound isolation workflow was guided by a molecular networking-based dereplication strategy. Twenty-three compounds were dereplicated from the EtOH extract of P. nitida and fractions network and were assigned various levels of identification confidence. The antiparasitic activities against Plasmodium falciparum as well as the cytotoxic activity against the MRC-5 cell line were determined for compounds 1-7.

Microscopic investigations and pharmacognostic techniques for the standardization of Caralluma edulis (Edgew.) Benth. ex Hook.f.

Herbal medicines frequently suffer with quality controversies because of similar species or varieties. This often leads to sophistication or admixture of the crude drug as they share various look alike physical features. Commercially, stalks of Caralluma edulis are commonly adulterated with other species because of their analogous morphology. In the present research, the microscopic standardization of the drug was performed by structural, histological, and anatomical features. Pharmacognostical characterization was carried out by physicochemical (moisture content, swelling index, pH value, ash values, and extractive values) and fluorescence analysis. The chemical characterization by UV-Visible, Fourier transform infrared spectroscopy, and high-performance liquid chromatography was also performed that disclosed the chief phytoconstituents of the herbal drug. The above mentioned constraints, being reported for the first time in C. edulis, are substantial in establishing the microscopic and pharmacopoeial standards for future identification and authentication.

Palynological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi, Pakistan, using light and scanning electron microscopy.

This was the first comprehensive pollen micromorphological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi, Pakistan. The pollen morphology of 10 species of the family Apocynaceae was observed and documented using light microscopy and scanning electron microscopy. Pollen was found subspheroidal in shape in most of the species, however peroblate shape was noted in Vinca major. Exine sculpturing patterns (psilate, rugulate, scabrate, and microreticulate) were observed. The result indicated that exine ornamentation of Apocynaceae taxa is systematically informative at generic and species levels. Most of the species have tricolporate type pollen but tetraporate pollen was also observed in Trachelospermum jasminoides and tetracolpate in V. major. Minimum equatorial diameter was noted in Carissa edulis (27.13 µm) and maximum in V. major (108.25 µm). Similarly, maximum exine thickness was found in Cascabela thevetia (9.5 µm). In the present findings, the pollen morphological data are compared with available other pollen studies to evaluate the taxonomic value of pollen traits in Apocynaceae taxa by using multiple microscopic techniques. Furthermore, molecular and phylogenetic studies were recommended to strengthen the systematics of Apocynaceae taxa.

Breakdown of species boundaries in Mandevilla: floral morphological intermediacy, novel fragrances and asymmetric pollen flow.

Phenotypic intermediacy is an indicator of putative hybrid origin and has provided the main clues to discovering hybrid plants in nature. Mandevilla pentlandiana and M. laxa (Apocynaceae) are sister species with clear differences in floral phenotype and associated pollinator guilds: diurnal Hymenoptera and nocturnal hawkmoths, respectively. The presence of individuals with intermediate phenotypes in a wild population raises questions about the roles of visual and olfactory signals (i.e. corolla morphology and floral fragrances) as barriers to interbreeding, and how the breakdown of floral isolation occurs. We examined phenotypic variation in a mixed Mandevilla population, analysing the chemical composition of floral fragrances, characterising floral shape through geometric morphometrics and assessing individual grouping through taxonomically relevant traits and an unsupervised learning algorithm. We quantified the visitation frequencies of floral visitors and tracked their foraging movements using pollen analogues. The presence of morphologically intermediate individuals and pollen analogue movement suggested extensive hybridisation between M. laxa and M. pentlandiana, along with asymmetrical rates of backcrossing between these putative hybrids and M. laxa. Floral volatiles from putative hybrid individuals showed a transgressive phenotype, with additional compounds not emitted by either parental species. Our results suggest the presence of a hybrid swarm between sympatric M. pentlandiana and M. laxa and indicate that initial hybridisation events between these parental species are rare, but once they occur, visits between putative hybrids and M. laxa are common and facilitate continued introgression.

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.

Evading plant defence: Infestation of poisonous milkweed fruits (Asclepiadaceae) by the fruit fly Dacus siliqualactis (Diptera: Tephritidae).

To cope with toxic metabolites plants use for defence, herbivorous insects employ various adaptive strategies. For oviposition, the fruit fly Dacus siliqualactis (Tephritidae) uses milkweed plants of the genus Gomphocarpus (Asclepiadaceae) by circumventing the plant's physical (gluey latex) and chemical (toxic cadenolides) defence. With its long, telescope-like ovipositor, the fly penetrates the exo- and endocarp of the fruit and places the eggs on the unripe seeds located in the centre of the fruit. Whereas most plant parts contain high concentrations of cardenolides such as gomphoside, calotropin/calacatin and gomphogenin, only the seeds exhibit low cardenolide levels. By surmounting physical barriers (fruit membranes, latex), the fly secures a safe environment and a latex-free food source of low toxicity for the developing larvae. One amino acid substitution (Q111V) at the cardenolide binding site of the fly's Na+, K+-ATPase was detected, but the significance of that substitution: reducing cardenolide sensitivity or not, is unclear. However, poisoning of the larvae by low levels of cardenolides is assumed to be prevented by non-resorption and excretion of the polar cardenolides, which cannot passively permeate the midgut membrane. This example of an insect-plant interaction demonstrates that by morphological and behavioural adaptation, a fruit fly manages to overcome even highly effective defence mechanisms of its host plant.

Evolution of the stapeliads (Apocynaceae-Asclepiadoideae) - repeated major radiation across Africa in an Old World group.

The stapeliads of the Ceropegieae (Apocynaceae-Asclepiadoideae), are approximately 340 species of stem-succulents placed in around 30 genera, found in semi-arid parts of the Old World. Here we sampled 192 species (i.e. nearly two thirds of the total) from across the full geographic range of the group and analysed data from the two nuclear regions (nuclear ribosomal ITS and ncpGS) and five plastid regions (psbA-trnH intergenic spacer, rps16 intron, trnL-trnF intergenic spacer, trnS-trnG intergenic region and the non-coding rpl32-trnL region). We find that the stapeliads radiated first in the northern hemisphere from Africa to southern Europe and Myanmar. This radiation subtends a grade of minor clades in the south-western corner of the African continent. These were followed by a single clade containing major radiation back across Africa from South Africa to tropical Arabia (but no further east than Dhofar, Oman), which includes also a single early spread into Madagascar. We establish the monophyly of many of the genera, such as Echidnopsis Hook.f., Hoodia Hook., Huernia R. Br., Piaranthus R. Br., Rhytidocaulon P.R.O. Bally and Tridentea Haw., but find that Duvalia Haw., Orbea Haw., Stapelia L. and Tromotriche Haw. are polyphyletic. We show that in certain vegetative features, there is broad cohesion across clades. Florally, on the other hand, the stapeliads exhibit considerable plasticity and we are able to show that very differently shaped flowers as well as large and small flowers evolved repeatedly among closely related species.

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.

Estimates of leaf vein density are scale dependent.

Leaf vein density (LVD) has garnered considerable attention of late, with numerous studies linking it to the physiology, ecology, and evolution of land plants. Despite this increased attention, little consideration has been given to the effects of measurement methods on estimation of LVD. Here, we focus on the relationship between measurement methods and estimates of LVD. We examine the dependence of LVD on magnification, field of view (FOV), and image resolution. We first show that estimates of LVD increase with increasing image magnification and resolution. We then demonstrate that estimates of LVD are higher with higher variance at small FOV, approaching asymptotic values as the FOV increases. We demonstrate that these effects arise due to three primary factors: (1) the tradeoff between FOV and magnification; (2) geometric effects of lattices at small scales; and; (3) the hierarchical nature of leaf vein networks. Our results help to explain differences in previously published studies and highlight the importance of using consistent magnification and scale, when possible, when comparing LVD and other quantitative measures of venation structure across leaves.

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.

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.

Pharmacobotanic characterization of young stems and stem barks of Rauvolfia sellowii Müll. Arg., Apocynaceae / Caracterização farmacobotânica de novos talos e cascas do caule de Rauvolfia sellowii Müll. Arg., Apocynaceae

Rauvolfia sellowii Müll. Arg. (Apocynaceae), a Brazilian native tree rich in indole alkaloids, is known as "pau-pra-tudo" and popularly used as hypocholesterolemic, hypoglycemic and antihypertensive. The aim of this work was to study the anatomy of the young stems and stem barks of this medicinal plant, in order to contribute to the identification of the species as a drug. The plant material was fixed and prepared according to standard microtechniques. The young stems have remaining epidermis, but a suberified peridermis is evident. The phellogen is located in the cortical region, forming suber externally. Underneath the phellogen, lies the phelloderm and collenchymatic region. In the cortex, there are numerous laticifers and some fibers. There is an incomplete sclerenchymatic sheath, consisting of several groups of fibers and stone cells. The stem has internal phloem ordered as isolated groups side by side. Numerous laticifers, calcium oxalate crystals, idioblasts and amyloplasts are found in the cortex, phloem, xylem and pith. The stem bark has many layers of suber and cortical parenchyma, a sheath composed of fibers and stone cells totally lignified, and external phloem. These anatomical characteristic, taken together, can be used as quality control parameters for this species.

Rauvolfia sellowii Müll. Arg. (Apocynaceae), uma árvore nativa brasileira rica em alcaloides indólicos, é conhecida como "pau-pra-tudo" e utilizada popularmente como hipocolesterolêmica, hipoglicêmica e anti-hipertensiva. O presente trabalho teve como objetivo caracterizar anatomicamente o caule jovem e a casca caulinar dessa planta medicinal, a fim de contribuir para a identificação e autenticidade da droga. O material vegetal foi fixado e submetido às microtécnicas usuais. O caule jovem possui epiderme remanescente, porém uma periderme suberificada é observada. O felogênio instala-se na região cortical, formando súber externamente. Subjacentes ao felogênio são observadas a feloderme e uma região colenquimática. No córtex, há numerosos laticíferos e algumas fibras. Há uma bainha esclerenquimática incompleta, constituída de vários grupos de fibras e células pétreas. O caule mostra floema interno organizado em grupos isolados lado a lado. Numerosos laticíferos, cristais de oxalato de cálcio, idioblastos e amiloplastos são encontrados no córtex, no floema, no xilema e na medula. A casca do caule possui várias camadas de súber e parênquima cortical, uma bainha composta de fibras e células pétreas totalmente lignificadas, e floema externo. Estes caracteres anatômicos, quando analisados em conjunto, podem ser utilizados como parâmetros para controle de qualidade desta espécie.

[First record of simuliidae (Diptera) with pollinaria of asclepiadoideae (Apocynaceae) attached]. / Primeiro registro de simuliidae (Diptera) com polinários de asclepiadoideae (Apocynaceae).

The presence of pollinaria of two species of Asclepiadoideae (Apocynaceae), possibly Tassadia cf. martiana Decne. and T. cf. obovata Decne., attached to the mouth parts of simulid black flies [Cerqueirellum amazonicum (Goeldi), C. argentiscutum (Shelley & Luna Dias), C. oyapockense (Floch & Abonnenc), and Cerqueirellum sp.] are reported for the first time. The frequency and distribution of simulids recorded with pollinaria suggest that removal of pollinaria by these flies is not casual. Simulids probably use nectar in flowers of Asclepiadoideae as source of sugar, being able to remove their pollinaria. This finding demonstrates that simulids are not only vector of pathogenic parasites, but also carry pollinaria, and thus may represent a group of pollinators for species of Asclepiadoideae with small flowers.

Morfo-anatomia da plântula de tabernaemontana catharinensis A. DC. (Apocynaceae) / Morphology and anatomy of the seedling of tabernaemontana catharinensis A. DC. (Apocynaceae)

Tabernaemontana catharinensis, conocida vulgarmente como lechero, es una especie perteneciente a la familia Apocynaceae. El actual estudio se refierea la morfología y la anatomía de la plántula de esta especie. Las semillas colectadas desde varios arboles creciendo en la planicie de la inundación del alto río Paraná, fueron germinadas en una cámara de germinación con una temperatura constante de 300C y mantenidas bajo luz fluorescente continua. El desarrollo de las plántulas ocurrió en un invernadero. La anatomía fue estudiada en plántulas fijadas en FAA 50. Este material fue seccionado transversalmente y las secciones obtenidas fueron teñidas en safranina y azul de astra. La plántula es fanerocotiledonar y epígea y presenta raíz axial, cuello, cotiledones ovados, epicótilo reducido, y eófilos lanceolados semejantes a los metáfilos. Su raíz es tetrarca. El hipocótilo y epicótilo tienen estructura caulinar y presentan crecimiento secundario cuya peridermis es de origen del felógeno en el ámbito epidérmico. Los cotiledones y eófilos son hojas dorsiventrales, con un estrato de parénquima en empalizada y algunas capas de parénquima esponjoso. Los cotiledones tienen un haz vascular colateral en la vena media, mientras en los eófilos el haz es bicolateral.

Tabernaemontana catharinensis is a species belonging to the family Apocynaceae. In this paper, the seedling morphology and anatomy of this species are described and discussed. The seedling development was analyzed until the first eophyll pair’s formation. The seeds were collected from several trees, in fragments of the semideciduous seasonal alluvial and submontaneforest located on the Upper Paraná River floodplain. The seeds were germinated at constant temperatures of 300C, using germination chambers and maintained under continuous fluorescent light. Seedlings development occurred in the greenhouse. The seedlings fixed in FAA 50 were sectioned by microtome and stained with safranin and astra blue. Seedling is phanerocotylar and epigeal with axial root, collet, ovate cotyledons, reduced epicotyl, and lanceolate eophylls similar to the metaphylls. Root is tetrarch. Hypocotyl and epicotyl have stem structure, and they present secondary growth with phellogen of epidermic origin. Cotyledons and eophylls are dorsiventral leaves, with uniseriate palisade parenchyma and some layers of spongy parenchyma. Cotyledons have a collateral vascular bundle in the midrib, while in the eophylls is bicollateral one.

Moth pollination of Metaplexis japonica (Apocynaceae): pollinaria transfer on the tip of the proboscis.

Asclepiad pollinaria (including pollen masses) attach to diverse body parts of flower visitors in many ways. In this paper, we observed nocturnal moths (Lepidoptera: Pyralidae and Noctuidae) transporting the pollinaria of the Japanese species Metaplexis japonica (Thunb.) Makino (Apocynaceae: Asclepiadoideae) on the tip of the proboscis. Flowers of this species may induce nectar-feeding moths to pull out the proboscis along a guide rail (anther slit), thus clipping the pollinaria onto the tip of the proboscis and transferring the pollinaria to the next flower. The transfer of pollinaria on the unique vector of a moth proboscis tip is an interesting pollination mechanism among previously reported entomophiles.

Morphological and ultrastructural diversity of orbicules in relation to evolutionary tendencies in apocynaceae s.L.

Minute granules of sporopollenin, called orbicules, can be observed on the innermost tangential and/or radial walls of secretory tapetum cells. Orbicules were investigated in 62 species (50 genera) of Apocynaceae s.l. using light microscopy, scanning electron microscopy and transmission electron microscopy. Orbicules were found in 43 species (34 genera) distributed amongst the subfamilies Rauvolfioideae, Apocynoideae, Periplocoideae, and in the genus Riocreuxia (Asclepiadoideae). Absence of orbicules is apparent in Secamonoideae and Asclepiadoideae (except Riocreuxia). The orbicule types described are based on observed morphological and ultrastructural variation. Of the six orbicule types previously described, Type I and Type II orbicules are lacking. In the majority of species, Type III orbicules were recorded in addition to Types IV, V and VI. In this study we suggest that embedded Type VI orbicules are more derived. A correlation between orbicule typology and evolutionary tendencies in Apocynaceae s.l. palynology was found. A trend was observed from the presence of Type III orbicules in the majority of species belonging to the basal group of genera characterized by colporate to porate single pollen grains, or 3-6-porate tetrads, towards the more derived embedded Type VI orbicules in the more advanced Periplocoideae genera with multiporate tetrads or pollinia. Orbicule data have proven not to be useful for evaluating tribal delimitation within the Apocynaceae s.l. contrary to the Rubiaceae and Loganiaceae s.l.

[The pharmacognostical identification of peel of Gymnema sylvestre].

This paper reported the descriptions, microscopic UV and TLC identification of peel of Gymnema sylvestre.