Anatomical and histochemical characterization of Dipteryx odorata and Taralea oppositifolia, two native Amazonian species

Abstract Dipteryx odorata (Aubl.) Willd. and Taralea oppositifolia Aubl., Fabaceae: Dipterygeae, are two Amazonian species of great economic and pharmacological potential. The anatomy of these species, however, remains poorly studied. The aim of this work was to inventory leaf anatomical characteristics of D. odorata and T. oppositifolia and to locate and identify secretory structures and determine the main classes of metabolites they store. Vegetative branches were collected in Parque Ecológico de Gunma, Belém, state of Pará, Brazil. Some of the branches were destined for herborization while the remainder was submitted to standard protocols for anatomical analysis and histochemical tests. Both species were found to possess an unstratified epidermis, with D. odorata being amphistomatic and T. oppositifolia being hypostomatic, and dorsiventral mesophyll with spongy parenchyma and wide cellular space. The two species were also found to possess idioblasts and secretory cavities that produce a heterogeneous exudate consisting of polysaccharides, lipids, alkaloids and phenolic compounds. The species presented differences in leaf anatomy and chemical composition of the secretory structures, which may be useful in their differentiation.

Drying temperature changes trichome integrity, chemical content and composition of the essential oil of pepper-rosmarin / Temperatura de secagem altera integridade de tricomas, teor e composição química doóleo essencial de alecrim-pimenta

ABSTRACT: Medicinal plants are generally commercialized dried. However, temperatures used in their drying processes may interfere with the content and chemical composition of their essential oils. The Lippia origanoides possesses thymol in the essential oil that is stored in glandular trichomes. Thymol is a major component of economic importance in the chemical and pharmaceutical industry. The objective of this study was to evaluate the effect of the drying temperatures of L. origanoides leaves regarding trichome integrity, content, and chemical composition of the essential oil. The experimental design was completely randomized with four treatments defined by oven drying temperatures (40, 50, 60, and 70ºC) and four replications. Essential oil was extracted by hydro distillation and the essential oil contents were determined (%). Chemical composition of the oil was analyzed by gas chromatography coupled to mass spectrometry. Scanning electron microscopy was carried out to determine trichome integrity. Drying the L. origanoides leaves in a forced ventilation oven at 40°C minimized the loss of extracted essential oil content (17.5g kg-1) and relative thymol percentage. Leaves dried at the temperatures of 60 and 70ºC exhibited a higher percentage of ruptured trichomes and reduced essential oil content to 13.7g kg-1 and 11.8g kg-1, respectively.

RESUMO: As plantas medicinais são geralmente comercializadas secas, no entanto, as temperaturas utilizadas nos processos de secagem podem interferir no teor e composição química dos óleos essenciais destas plantas. A Lippia origanoides possui timol em seu óleo essencial, que se encontra armazenado em tricomas glandulares. O timol é um componente majoritário de importância econômica na indústria química e farmacêutica. O objetivo do trabalho foi estudar o efeito das temperaturas de secagem de folhas de L. origanoides na integridade dos tricomas, teor e composição química do óleo essencial. O delineamento experimental foi inteiramente casualizado, com quatro tratamentos definidos por temperaturas de secagem em estufa (40ºC, 50ºC, 60ºC e 70ºC) e quatro repetições. Foram realizadas extrações por hidrodestilação e determinação dos teores dos óleos essenciais (%), análise de composição química do óleo por cromatografia gasosa acoplada à espectrometria de massas e microscopia eletrônica de varredura para determinação da integridade dos tricomas. A secagem das folhas de L. origanoides em estufa de ventilação forçada a 40ºC minimizou a perda do teor de óleo essencial extraído (17,5g kg-1) e porcentagem relativa de timol. Folhas secas em temperatura de 60 e 70ºC exibiram maior porcentagem de tricomas rompidos e redução do teor de óleo essencial para 13,7g kg-1 e 11,8g kg-1, respectivamente.

Morphoanatomical and physicochemical profile of Piper callosum: valuable assessment for its quality control

ABSTRACT Piper callosum Ruiz & Pav., Piperaceae, popularly known as “elixir-paregórico” and “matricá” in Brazil, is used in folk medicine to treat gonorrhea, general pain, and digestive disorders, and has repellent, astringent, diuretic, depurative, and haemostatic properties. Despite the fact that this plant is sold as a traditional phytotherapeutic product, we did not find reports on its quality control. We, therefore, performed macroscopic, microscopic, histochemical, and physicochemical analyses using standard methods to establish botanical authentication and purity degree parameters for leaves and stem of this species in two forms: medicinal plant and herbal drug. We observed the size, shape, color, texture, fracture surface and transection characteristics, leaf venation patterns, and calluses are valuable diagnostic characters to identify the herbal drugs when they are not ground or powdered. Since medicinal plants and herbal drugs did not differ anatomically, the following key anatomical characters for P. callosum can be used for diagnostic purposes of both types raw plant materials: epicuticular wax and cuticular flanges patterns; collenchyma features; fibers in the midrib; arrangement pattern of the vascular bundles of the midrib and petiole; shape of the midrib, leaf margin, petiole, and stem; occurrence of raphides; and morphology of the starch grains. Acid lipids, essential oils, oleoresins, steroids, tannins and flavonoids were histochemically identified. Total ash (leaves: 11.25%; stem: 5.25%), sulphated ash (leaves: 68.02%; stem: 12.50%), acid-insoluble ash (leaves: 2.82%; stem: 0.27%), moisture (leaves: 8.60%; stem: 6.10%), loss on drying (leaves: 11.08%; stem: 8.58%), and pH (leaves: 5.57, stem: 5.28) values were determined. The order of analyzed metal levels in leaf and stem herbal drugs was Al > V > Cu > Mn > Cr > Ni. Similar levels of Cd and Co and low levels of Hg were found. The results obtained can be used as quality control parameters for medicinal plants and herbal drugs of P. callosum.

Structure and distribution of glandular trichomes in three species of Bignoniaceae / Estrutura e distribuição de tricomas glandulares em três espécies da família Bignoniaceae

Glandular trichomes play a major role in the morphological characterization of the Bignoniaceae. Due to their great diversity of forms and functions, this study aimed to inventory the glandular trichomes present in the aerial vegetative axis of Amphilophium magnoliifolium, Martinella obovata and Stizophyllum riparium, analyze their structure and register the participation of ants in these plants. Fresh samples from the nodal region, petiole and from medium to apical regions of the leaflet blade were fixed and processed according to usual methods in light and scanning electron microscopies. The glandular trichomes found were: peltate, capitate, stipitate, and patelliform/cupular. Peltate trichomes are the most abundant ones and present the most uniform distribution. Patelliform/cupular trichomes occur at specific regions, such as prophylls, leaflet blade and nodal regions. Martinella obovata is the only species that presents capitate and stipitate trichomes, which are widely distributed along the entire aerial vegetative axis. Ants were found in all species, mainly at nodal regions. The occurrence of the capitate-type trichome is reported for the first time to the genus.

Tricomas glandulares desempenham papel importante na caracterização morfológica de Bignoniaceae e, devido à grande diversidade de formas e funções que esses tricomas apresentam, o objetivo desse estudo foi inventariar os tricomas glandulares presentes no eixo vegetativo aéreo de Amphilophium magnoliifolium, Martinella obovata. e Stizophyllum riparium, bem como analisar sua estrutura e registrar a participação de formigas nessas espécies. Amostras frescas da região mediana à apical das lâmina foliolar, pecíolo e nó foram fixadas e processadas de acordo com os métodos usuais para microscopia de luz e eletrônica de varredura. Os tricomas glandulares encontrados nas espécies foram: peltados, capitados, estipitados e pateliformes/cupuliformes. Dentre esses tipos o mais abundante foi o tricoma peltado, apresentando também uma distribuição mais uniforme que os demais. Os tricomas pateliformes/cupuliformes estavam em regiões mais específicas, como profilos, lâmina foliolar e na região nodal. Martinella obovata foi a única espécie que apresentou os tricomas capitados e estipitados, amplamente distribuídos ao longo de todo eixo vegetativo aéreo. Em todas as espécies foram encontradas formigas, principalmente nas regiões nodais. A ocorrência do tricoma capitado é relatada pela primeira vez para o gênero.

Morphological and micromorphological characteristics of Desmodium fruits (Leguminosae: Papilionoideae)

The genus Desmodium is represented in Santa Catarina State, Brazil, by 13 species, all with lomentaceous fruits. Shape, size and isthmus margin of loments vary, while the surface is glabrous, or covered by trichomes of different types. Morphological diversity of trichomes becomes particularly relevant to taxonomic description. The trichome types present on the surface of Desmodium fruits provide data for the identification and classification of species in the State. To assess this, three fruits of each species were collected and deposited at two herbaria, HBR and FLOR, in Santa Catarina, Brazil. Some rehydrated samples were examined using light microscopy (LM); and some sections were exposed to the following histochemical reagents: Sudan III for oils and Thionine for mucilage. The structural aspects of trichomes can be classified into uni- or multicel- lular and may still be simple, i.e., nonglandular or glandular. Using scanning electron microscopy (SEM), five types of trichomes were identified and analyzed among the Desmodium species studied: uncinate, uniseriate, globose multicellular, globose unicellular and subulate. Characteristics, such as loment margin and article form, glabrescent or pillous indument, trichome type, with or without papillous epidermal cells and epicuticular striations, showed relevant diagnostic value. An identification key was developed for Desmodium species from Santa Catarina State, Brazil, based on macro and micromorphological characters of the fruit.

Características morfológicas y micromorfológicos de frutos Desmodium (Leguminosae: Papilionoideae). El género Desmodium está representado en Santa Catarina, Brasil, por 13 especies, todas con frutos lomentaceos. Los lomentos han tenido variación en forma, tamaño y características del margen del istmo, y la superficie es glabra o cubierta por tricomas de diferentes tipos. La diversidad morfológica de los tricomas se vuelve particularmente relevante para la descripción taxonómica. Los tipos de tri- comas presentes en la superficie de los frutos Desmodium, proporcionan datos para la identificación y clasificación de las especies en el Estado de Santa Catarina, Brasil. Tres frutos de cada especie fueron recogidos y depositados en el Herbario (HBR y FLOR, Santa Catarina, Brasil). Algu- nas muestras fueron rehidratadas y examinadas usando microscopía de luz (LM), las secciones fueron expuestas a los siguientes reactivos histoquímicos: Sudan III para los aceites y Tionina para mucílago. Los aspectos estructurales de los tricomas se pueden clasificar en uni o multicelulares y pueden todavía ser simples, es decir, no glandular o glandular. Por medio del uso de la microscopía elec- trónica de barrido (SEM), cinco tipos de tricomas se han identificado y analizado entre las especies de Desmodium estudiadas: uncinado, uniseriado, globoso multicelular, globoso unicelular y subulado. Características como el margen del lomento y la forma del artículo, indumento glabrescente o piloso, tipo de tricomas con o sin células epidérmicas papilosas, y estrías epicuticulares mostraron valor de diagnóstico relevante. La clave de identificación fue desarrollada para especies de Desmodium del Estado de Santa Catarina, Brasil, basado en caracteres macro y micromorfológicos del fruto.

Chemical composition and histochemistry of Sphagneticola trilobata essential oil

Anatomical and histochemical investigations of Sphagneticola trilobata (L.) Pruski, Asteraceae, secretory structures in leaves and stems and the seasonal variation of essential oils were carried out. Histochemical techniques enabled the specific location of the essential oil accumulation in the internal (canals) and external structures (trichomes). Histochemical analysis showed that the secretory trichomes produced steroids. The highest yield was obtained from plants collected in winter, when it was registered low temperature and precipitation. The essential oil was characterized by high percentage of hydrocarbon sesquiterpenes, hydrocarbon monoterpenes and low levels of oxygenated sesquiterpenes. The major components were germacrene D (11.9-35.8%), α-phellandrene (1.4-28.5%), α-pinene (7.3-23.8%), E-caryophyllene (4.6-19.0%), bicyclogermacrene (6.0-17.0%), limonene (1.8-15.1%) and α-humulene (4.0-11.6%). The percentage of most of the individual constituents present in S. trilobata essential oil changed significantly during the months.

Anatomia e histoquímica dos órgãos vegetativos de Polygonum hydropiperoides Michx., Polygonaceae / Anatomical and histochemical characters of Polygonum hydropiperoides Michaux, Polygonaceae

Polygonum hydropiperoides Michaux é uma espécie conhecida popularmente como "erva-de-bicho", amplamente utilizada na medicina tradicional como anti-hemorroidal, antiinflamatória e antidiarréica. O presente trabalho tem como objetivo a caracterização anatômica e histoquímica da folha, caule e raiz, que constituem a droga vegetal, visando estabelecer parâmetros para o controle de qualidade. O material vegetal foi fixado e submetido às técnicas usuais de microscopia de luz e aos testes histoquímicos. A folha é anfiestomática, dorsiventral e com estômatos paracíticos e anisocíticos. É comum a presença de estruturas secretoras como: tricomas glandulares capitados e glândulas epidérmicas e subepidérmicas em ambas as faces da lâmina foliar e também no caule. O material secretado pelas glândulas apresenta composição heterogênea de lipídios e flavonóides, segundo análises histoquímicas. Cristais de oxalato de cálcio e grãos de amido são freqüentes em células parenquimáticas da folha e do caule. Compostos fenólicos estão presentes na folha (parênquima paliçádico), no caule (parênquima cortical e floema) e na raiz (parênquima cortical).

P. hydropiperoides is popularly known as "erva-de-bicho", and it is used in traditional medicine as anti-hemorroidal, anti-inflammatory and antidiarrhoeic. The present work has as objective the anatomical and histochemical characterization of leaf, stem and root that are used as drug in order to offer elements for quality control. The botanical material was prepared for the usual optical and histochemical microtechniques. The leaf is amphistomatic, dorsiventral and paracytic and anisocytic stomata. It is common to find the presence of secretory structures as: capitate glandular trichomes and secretory glands on both epidermis, that are also present in stem. The secretion material is heterogeneous composition of lipids and flavonoids. Druses of calcium oxalate and starch are common in parenchyma cells of leaf and stem. Phenolic compounds are present in leaf (palisade parenchyma), in stem (cortical parenchyma and phloem) and root (cortical parenchyma).