Phytochemical study guided by the myorelaxant activity of the crude extract, fractions and constituent from stem bark of Hymenaea courbaril L.

ETHNOPHARMACOLOGICAL RELEVANCE: Hymenaea courbaril L. (Caesalpinoideae) is used in Brazilian folk medicine to treat anemia, kidney problems, sore throat and other dysfunctions of the respiratory system, such as bronchitis and asthma, although such properties are yet to be scientifically validated. AIM OF THE STUDY: In order to give a scientific basis to support the traditional use of Hymenaea courbaril, this study was designed to evaluate antioxidant, myorelaxant and anti-inflammatory properties of the ethanol extract from stem bark and its fractions. The myorelaxant effect of astilbin, a flavonoid isolated from the bioactive ethyl acetate fraction (EAF), has also been evaluated. MATERIAL AND METHODS: In the present study ethanol extract from stem bark (EEHC) and fractions were analyzed using bioassay-guided fractionation. The following activities were investigated: antioxidant by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, myorelaxant on rat tracheal smooth muscle, and anti-inflammatory using ovalbumin-induced leukocytosis and airway hyperresponsiveness in rats. RESULTS: The results of the present investigation show that the whole extract of Hymenaea courbaril and some of its fractions strongly scavenged DPPH radical. The extract showed myorelaxant activity on rat trachea, being EAF its highest efficient fraction. Bio-guided study allowed the isolation of astilbin, a well-known flavonoid. The activity induced by this compound indicates that it may be partly responsible for the myorelaxant effect of EAF. EAF reduced contractions that depended on divalent cation inflow through voltage-operated Ca(2+) channels (VOCCs) or receptor-operated Ca(2+) channels (ROCCs), but it was more potent to inhibit VOCC- than ROCC-dependent contraction induced by Ca(2+) addition in ACh-enriched Ca(2+)-free medium. Oral pretreatment of antigen-challenged animals with EAF prevented airway hyperresponsiveness on KCl-induced contraction and reduced the number of total white cells, particularly eosinophils and neutrophils in bronchoalveolar lavage. CONCLUSIONS: This study provided scientific basis that Hymenaea courbaril presents potential antioxidant, myorelaxant and anti-inflammatory actions, which support its use in folk medicine to treat inflammatory airway diseases.

Mendelian inheritance, linkage and genotypic disequilibrium in microsatellite loci isolated from Hymenaea courbaril (Leguminosae).

The Neotropical tree Hymenaea courbaril, locally known as Jatobá, is a valuable source of lumber and also produces comestible and medicinal fruit. We characterized Mendelian inheritance, linkage and genotypic disequilibrium at nine microsatellite loci isolated from H. courbaril, in order to determine if they would provide accurate estimates of population genetic parameters of this important Amazon species. The study was made on 250 open-pollinated offspring originated from 14 seed trees. Only one of nine loci presented significant deviation from the expected Mendelian segregation (1:1). Genotypic disequilibrium between pairwise loci was investigated based on samples from 55 adult and 56 juvenile trees. No genetic linkage between any paired loci was observed. After Bonferroni's corrections for multiple tests, we found no evidence of genotypic disequilibrium between pairs of loci. We conclude that this set of loci can be used for genetic diversity/ structure, mating system, gene flow, and parentage analyses in H. courbaril populations.

Do tropical wetland plants possess convective gas flow mechanisms?

• Internal pressurization and convective gas flow, which can aerate wetland plants more efficiently than diffusion, are common in temperate species. Here, we present the first survey of convective flow in a range of tropical plants. • The occurrence of pressurization and convective flow was determined in 20 common wetland plants from the Mekong Delta in Vietnam. The diel variation in pressurization in culms and the convective flow and gas composition from stubbles were examined for Eleocharis dulcis, Phragmites vallatoria and Hymenachne acutigluma, and related to light, humidity and air temperature. • Nine of the 20 species studied were able to build up a static pressure of > 50 Pa, and eight species had convective flow rates higher than 1 ml min(-1). There was a clear diel variation, with higher pressures and flows during the day than during the night, when pressures and flows were close to zero. • It is concluded that convective flow through shoots and rhizomes is a common mechanism for below-ground aeration of tropical wetland plants and that plants with convective flow might have a competitive advantage for growth in deep water.

Crescimento de Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell) Morong (Leguminosae) sob diferentes níveis de sombreamento / Growth of Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell) Morong (Leguminosae) under different shading levels

Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. é uma espécie clímax tolerante a sombra, ao passo que Enterolobium contortisiliquum (Vell.) Morong. é uma espécie pioneira. O desenvolvimento destas espécies pode refletir a habilidade de adaptação aos diferentes fatores ambientais (luz, água e temperatura) no local em que estão crescendo. O suprimento inadequado de um desses fatores pode reduzir o vigor da planta e limitar seu desenvolvimento. O presente trabalho teve como objetivo avaliar os efeitos do nível de sombreamento no crescimento e a concentração de pigmentos fotossintéticos em duas espécies de leguminosas arbóreas, Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell.) Morong. O experimento foi conduzido no Setor de Olericultura do Centro Universitário Luterano de Ji-Paraná (CEULJI/ULBRA)/Rondônia. Durante a formação das mudas, ambas as espécies foram expostas a quatro tratamentos de sombra: 0 % (controle - sol pleno); 30 %; 50 % e 80 %. Cada tratamento foi constituído com três repetições de cada espécie; o delineamento experimental foi inteiramente casualisado. Quatro meses após a semeadura, as seguintes análises foram realizadas: número de folhas, altura da planta, comprimento do sistema radicular, massa seca total e concentração de pigmentos fotossintéticos. O tratamento sob sol pleno afetou negativamente o crescimento de ambas as espécies. As mudas crescidas sob 50% e 80% apresentaram melhor desenvolvimento. Conforme o aumento do sombreamento houve um decréscimo na razão clorofila a/b e um aumento nas concentrações de clorofila total e carotenóides totais.

Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. is a clímax shadow tolerant specie and Enterolobium contortisiliquum (Vell.) Morong., by the other hand, is considered as a pioneer specie. The development of these species may reflect its adaptation ability to different environmental elements (light, water and temperature) at its growth site. An inadequate supply of some of these elements may reduce plant vigor and limit its development. This work aimed to evaluate the effects of shade intensity on the growth and photosynthetic pigment concentration of two arboreal legumes that are typical to northern Brazil, namely, the Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. and the Enterolobium contortisiliquum (Vell.) Morong. The experiment was carried out at the Centro Universitário Luterano de Ji-Paraná (CEULJ/ULBRA)/Rondônia. During the formation of the seedlings, both species were exposed to four treatments of shade: 0 % (control - without shade); 30 %; 50 % e 80 %. Each treatment was carried out with three replications for each species; the experimental design was completely randomized. Four month after sowing, the following analyses were carried out: number of leaves, plant height, roots length, dry mass and photosynthetic pigment concentration. The treatment under 0% shade impaired the seedling growth of both species. The most favorable treatments were 50 % and 80 % shade. According to the shade intensity, the chlorophyll a/b ratio decreased and the chlorophyll and carotenoids increased.

Expression pattern of four storage xyloglucan mobilization-related genes during seedling development of the rain forest tree Hymenaea courbaril L.

During seedling establishment, cotyledons of the rain forest tree Hymenaea courbaril mobilize storage cell wall xyloglucan to sustain growth. The polysaccharide is degraded and its products are transported to growing sink tissues. Auxin from the shoot controls the level of xyloglucan hydrolytic enzymes. It is not yet known how important the expression of these genes is for the control of storage xyloglucan degradation. In this work, partial cDNAs of the genes xyloglucan transglycosylase hydrolase (HcXTH1) and beta-galactosidase (HcBGAL1), both related to xyloglucan degradation, and two other genes related to sucrose metabolism [alkaline invertase (HcAlkIN1) and sucrose synthase (HcSUS1)], were isolated. The partial sequences were characterized by comparison with sequences available in the literature, and phylogenetic trees were assembled. Gene expression was evaluated at intervals of 6 h during 24 h in cotyledons, hypocotyl, roots, and leaves, using 45-d-old plantlets. HcXTH1 and HcBGAL1 were correlated to xyloglucan degradation and responded to auxin and light, being down-regulated when transport of auxin was prevented by N-1-naphthylphthalamic acid (NPA) and stimulated by constant light. Genes related to sucrose metabolism, HcAlkIN1 and HcSUS1, responded to inhibition of auxin transport in consonance with storage mobilization in the cotyledons. A model is proposed suggesting that auxin and light are involved in the control of the expression of genes related to storage xyloglucan mobilization in seedlings of H. courbaril. It is concluded that gene expression plays a role in the control of the intercommunication system of the source-sink relationship during seeding growth, favouring its establishment in the shaded environment of the rain forest understorey.

Diversity of regenerating plants and seed dispersal in two canopy trees from Colombian Amazon forests with different hunting pressure

Hunting may have drastic effects on some populations of frugivores and seed dispersal agents, which in turn may affect patterns of forest regeneration and plant diversity. However, when a species disappears, it is possible that the population of other species increases due to competition release, compensating or not, their ecological roles. The main aim of this study was to measure density compensation in primate communities, their possible effects on seedling and sapling diversity, and compensation of seed removal rates in two canopy trees (Apeiba aspera and Hymenaea oblongifolia). The study site was Amacayacu National Park (Colombian Amazon), where we carried out a census of mammal species using line transect methods in two forests with different levels of hunting pressure. Vegetation plots were used to quantify plant diversity for seedlings and saplings (4 m² and 25 m² respectively), and fruit traps were set up to estimate seed removal of A. aspera and H. oblongifolia. Large primates were less frequently encountered near human settlements, while small primates exhibited the opposite pattern, suggesting a density compensation effect. The diversity of regenerating plants was higher in the forest where large primates occur. Seed removal was higher in forests with large primates for A. aspera, but not for H. oblongifolia. Overall, the results support the hypothesis of density compensation on the primate community; however, there is no strong evidence of ecological compensation in terms of seed dispersal and regeneration in the species studied. Finally, the presence of large seed dispersers (e.g. Lagothrix lagothricha) was associated with higher plant diversity. Rev. Biol. Trop. 56 (3): 1531-1542. Epub 2008 September 30.

La cacería ha tenido fuertes efectos en algunas poblaciones de frugívoros y dispersores de semillas, lo cual en consecuencia puede afectar los patrones de diversidad de plantas y regeneración del bosque. Sin embargo, es posible que extinciones locales promuevan el incremento de otras especies, que pueden o no compensar roles ecológicos. El principal objetivo de este estudio fue examinar la posibilidad de compensación por densidad en las comunidades de primates y sus posibles efectos sobre la diversidad de plántulas y juveniles, así como también la remoción de semillas de dos árboles (Apeiba aspera e Hymenaea oblongifolia). El trabajo fue realizado en el Parque Nacional Natural Amacayacu (Amazonas - Colombia). Hicimos censos de mamíferos usando transectos lineales en dos bosques con diferentes presiones de cacería. Además, levantamos parcelas de vegetación para plántulas y juveniles (4 m² y 25 m² respectivamente), y ubicamos trampas de frutos bajo al menos seis árboles de cada especie, en cada bosque, para estimar la remoción de semillas. Los encuentros con primates grandes fueron menos frecuentes en bosques cerca de las comunidades indígenas, mientras que para los primates pequeños el patrón fue contrario, sugiriendo un efecto de compensación por densidad. Por otra parte, la diversidad de plántulas y juveniles fue más alta en bosques donde la densidad de primates grandes es mayor. La remoción de semillas fue mayor en bosques con mayor densidad de primates grandes para A. aspera, pero para H. oblongifolia las diferencias entre bosques no fueron significativas. En general, los resultados de este estudio apoyan la hipótesis de compensación por densidad en la comunidad de primates, sin embargo, no hay evidencia fuerte de compensación ecológica en términos de dispersión de semillas y regeneración. Finalmente, la presencia de dispersores grandes (ej. Lagothrix lagothricha) estuvo asociada con una mayor diversidad de plantas.

Diversity of regenerating plants and seed dispersal in two canopy trees from Colombian Amazon forests with different hunting pressure.

Hunting may have drastic effects on some populations of frugivores and seed dispersal agents, which in turn may affect patterns of forest regeneration and plant diversity. However, when a species disappears, it is possible that the population of other species increases due to competition release, compensating or not, their ecological roles. The main aim of this study was to measure density compensation in primate communities, their possible effects on seedling and sapling diversity, and compensation of seed removal rates in two canopy trees (Apeiba aspera and Hymenaea oblongifolia). The study site was Amacayacu National Park (Colombian Amazon), where we carried out a census of mammal species using line transect methods in two forests with different levels of hunting pressure. Vegetation plots were used to quantify plant diversity for seedlings and saplings (4 m2 and 25 m2 respectively), and fruit traps were set up to estimate seed removal of A. aspera and H. oblongifolia. Large primates were less frequently encountered near human settlements, while small primates exhibited the opposite pattern, suggesting a density compensation effect. The diversity of regenerating plants was higher in the forest where large primates occur. Seed removal was higher in forests with large primates for A. aspera, but not for H. oblongifolia. Overall, the results support the hypothesis of density compensation on the primate community; however, there is no strong evidence of ecological compensation in terms of seed dispersal and regeneration in the species studied. Finally, the presence of large seed dispersers (e.g. Lagothrix lagothricha) was associated with higher plant diversity.

The floral nectary of Hymenaea stigonocarpa (Fabaceae, Caesalpinioideae): structural aspects during floral development.

BACKGROUND AND AIMS: Considering that few studies on nectary anatomy and ultrastructure are available for chiropterophilous flowers and the importance of Hymenaea stigonocarpa in natural 'cerrado' communities, the present study sought to analyse the structure and cellular modifications that take place within its nectaries during the different stages of floral development, with special emphasis on plastid dynamics. METHODS: For the structural and ultrastructural studies the nectary was processed as per usual techniques and studied under light, scanning and transmission electron microscopy. Histochemical tests were employed to identify the main metabolites on nectary tissue and secretion samples. KEY RESULTS: The floral nectary consists of the inner epidermis of the hypanthium and vascularized parenchyma. Some evidence indicates that the nectar release occurs via the stomata. The high populations of mitochondria, and their juxtaposition with amyloplasts, seem to be related to energy needs for starch hydrolysis. Among the alterations observed during the secretory phase, the reduction in the plastid stromatic density and starch grain size are highlighted. When the secretory stage begins, the plastid envelope disappears and a new membrane is formed, enclosing this region and giving rise to new vacuoles. After the secretory stage, cellular structures named 'extrastomatic bodies' were observed and seem to be related to the nectar resorption. CONCLUSIONS: Starch hydrolysis contributes to nectar formation, in addition to the photosynthates derived directly from the phloem. In these nectaries, the secretion is an energy-requiring process. During the secretion stage, some plastids show starch grain hydrolysis and membrane rupture, and it was observed that the region previously occupied by this organelle continued to be reasonably well defined, and gave rise to new vacuoles. The extrastomatic bodies appear to be related to the resorption of uncollected nectar.