Occurrence of secretory structures in underground systems of seven Asteraceae species

In contrast with the abundance of anatomical studies of secretory structures on aerial vegetative organs of Asteraceae species, the information about secretory structures on thickened subterranean organs is sparse. The aim of this study was to investigate the occurrence of secretory structures on thickened and nonthickened subterranean organs of seven Asteraceae species from three tribes: Eupatorieae (Chromolaena squalida and Gyptis lanigera), Vernonieae (Chresta sphaerocephala, Lessingianthus bardanoides, L. glabratus and Orthopappus angustifolius), and Plucheeae (Pterocaulon angustifolium). The specimens were collected in areas of cerrado, from the State of Sao Paulo, Brazil. All species of the tribe Vernonieae studied exhibited endodermic cells, other than the epithelial cells of the canal, with secretory activity in the roots. In C. sphaerocephala roots, two types of endodermic cell were found, but only one had secretory activity. Secretory canals were found in the tuberous and nontuberous roots of all studied species. These data agree with the results from the literature for Asteraceae species. Here, we describe for the first time in Asteraceae the presence of secretory idioblasts in C. sphaerocephala. Secretory trichomes are present in the Orthopappus angustifolius rhizophore. Histochemical tests have shown that all types of secretory structure possess substances containing lipids. (C) 2008 The Linnean Society of London.

Fruit anatomy of Neotropical species of Indigofera (Leguminosae, Papilionoideae) with functional and taxonomic implications

LEITE, V. G., F. S. MARQUIAFAVEL, D. P. MORAES, AND S. P. TEIXEIRA (Departamento de Ciencias Farmaceuticas, Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, Universidade de Sao Paulo (USP), Av. do Cafe, s/n, 14040-903 Ribeirao Preto, SP, Brazil). Fruit anatomy of Neotropical species of Indigofera (Leguminosae, Papilionoideae) with functional and taxonomic implications. J. Torrey Bot. Soc. 136: 203-211. 2009-This work reports on the fruit surface and anatomy of seven Neotropical species of Indigofera (I. campestris Bong. ex Benth., I. hirsuta L., I. lespedeziodes Kunth, I. microcarpa Desv., I. spicata Forssk., I. suffruticosa Mill., and I. truxillensis Kunth) to help species diagnosis and clarify the fruit type classification. Flowers and fruits at several stages of development were removed from living material, fixed, and examined with scanning electron (surface analyses) and light microscopies (histological analyses). Species showed differences in relation to the number of exocarp layers, secretory trichome morphology and distribution, presence of stomata, phenolic idioblast size and distribution in mesocarp, the number and arrangement of endocarp fibers, and the presence of it separation tissue. It is noteworthy that no separation tissue was observed in L microcarpa and I. suffruticosa, although they have dehiscent fruits, which indicates it delayed dehiscence. The present work confirms that fruit anatomical characters can be utilized as it tool for fruit type classification, especially in Indigofera, the third largest genus of Leguminosae.

Identification of intracellular calcium oxalate crystals in Chamelaucium uncinatum (Myrtaceae)

Intracellular inclusions in the pedicel and calyx-tube tissues of Chamelaucium uncinatum Schauer ( Myrtaceae) flowers are irregular in shape. They were shown, by polarised light and scanning electron microscopy, to be birefringent 8.9-29.5 mum druse (i.e. aggregate) crystals. Energy-dispersive X-ray spectroscopy showed that these crystals were predominantly composed of calcium. Histochemical and acid-solubility tests indicated that the crystals were calcium oxalate. Raman microprobe spectroscopy was used to confirm this chemical identity. The calcium oxalate crystals were located in xylem-vessel lumens and also in parenchyma cells adjacent to vascular tissues. Thus, the crystals may function to regulate soluble calcium concentrations in C. uncinatum tissues near sites where calcium is unloaded from the xylem.

Morphology, ontogeny and structure of the stipular nectaries in Caamembeca spectabilis (Polygalaceae)

ABSTRACT Nodal glands are found in one third of the Polygalaceae genera and have valuable taxonomic, ecological and evolutionary significance. In Brazil, they occur in five of the eleven genera already registered. However, there is still a controversy regarding the origin of these structures. The objective of this study was to characterize the morphology and the origin of nodal glands inCaamembeca spectabilis, in order to increase the structural and functional knowledge of these glands in the genera. Samples of nodal regions were collected, fixed and processed according to the methods of light microscopy and electron scanning. Ants were observed and identified along the stem axis. The glucose in exudate allows us to classify these glands as extrafloral nectaries. They are located in pairs on the nodal region. However, its origin is in the leaf trace. In the longitudinal section, the nectaries were present in the apex of cells with anticlinal walls impregnated with suberin, which represents the first record for the family. In this region there is also the formation of a hole by lysis. The secretory tissue is surrounded by phloem. Xylem vessels were observed only on the basis of the nectary, where there are also idioblasts with crystals in druse type. We have studied the ontogeny of the glands nodal in Caamembeca spectabilis and unveiled that these glands are linked to the leaves as stipular nectaries. In addition, the new findings presented here may add support for the understanding of morphology and anatomy of nodal glands in Caamembeca.

Foliar anatomy of the species Lantana camara and L. radula (Verbenaceae)

The species Lantana camara, commonly used as ornamental, has spread worldwide becoming one of the world's most important weeds. To develop new methods of control of this plant, it is essential to distinguish it from other species of the same genus, and this is usually accomplished through taxonomic studies of fertile samples. Considering the similarity between L. camara and L. radula, and the consequent difficulty in distinguishing one from the other when only sterile samples are available, this work aimed to investigate the use of the anatomical characteristics of the leaves of both species as tools for supporting correct classification. The leaves of L. camara and L. radula were anatomically examined by light microscopy and scanning electron microscopy. The major differences were observed in the petiole, which presented secretory idioblasts in L. camara. Secretory idioblasts were observed in the leaf blades of L. camara and Crystalliferou idioblasts were found in L. radula. Glandular and nonglandular trichomes as well as the abaxial surface are different in each species. Such results can support the strategies aiming at the control of L. camara without interfering with L. radula.

Ontogeny and structure of the pericarp and the seed coat of Miconia albicans (Sw.) Triana (Melastomataceae) from " cerrado" , Brazil

Miconia albicans fruit and seed coat ontogeny were described under light microscope. The samples were fixed in formalin-aceto-alcohol (FAA), neutral-buffered formaldehyde solution (NBF) and formalin-ferrous sulphate (FFS) solutions, embedded in plastic resin, sectioned at 10 µm and stained with Toluidine Blue. Specific dyes and/or reagents were used for the microchemical tests. The ovary is semi-inferior and the indehiscent, fleshy globose berries are originated mainly from the development of the inferior portion of the ovary. The immature pericarp is mainly parenchymatous with some sclereids, druse crystal and phenolic-like compounds idioblasts widespread in the mesocarp. In the mature pericarp, the endocarp cells are often collapsed, the mesocarp is thick with cells more or less turgid, and the sclereids, the druses and the phenolic-like compound idioblasts are almost absent. The ovules are anatropous, bitegmic and crassinucellate, and the zig-zag micropyle is formed by both the exostome and the endostome. The mature seed is pyramidal-elongated in shape, exalbuminous and testal. The raphal part occupies about 40% of the seed coat total length and had the mechanical layer derived from its inner layer. The antiraphal side is non-multiplicative and the exotesta, mesotesta and endotesta are differentiated into a sclerotic layer, with the exotesta being the mechanical one. The tegmen is absent.

Polyphenolic cells and their interrelation with cotyledon cells in seven species of Theobroma (Sterculiaceae)

The amount of cotyledon polyphenolic cells varies extensively within the Theobroma species. The polyphenolic compounds of these cells play a protective role and furthermore have an important function in the development of chocolate flavour. The morphology of the polyphenolic cells of the mesophyll is described and the development of these idioblasts in Theobroma cacao L., T. subincanum Mart., T. obovatum Klotzsch ex Bernoulli, T. grandiflorum (Willd. ex Spreng.) K. Schum., T. microcarpum Mart., T. bicolor Bonpl. and T. speciosum Willd. ex Spreng analysed. The total polyphenolic content in the seeds as determined by spectrophotometry showed a variation of about forty times. The alive, transparent polyphenolic cells are scattered throughout the cotiledonary mesophyll. However the polyphenolic cells of T. cacao and T. grandiflorum are also aligned perpendicularly with respect to the mesophyll borders and, in addition, both species display polyphenolic cells with a natural translucent purple colour. All the species analysed contained polyphenolic cells scattered throughout the parenchymal cells and also in a lengthwise association with vascular bundles. In T. bicolor and T. speciosum, the species with the lowest polyphenolic contents, these cells were mostly located around the vascular bundles. Using Scanning Electron Microscopy, the polyphenolic cells demonstrated a complex cytoarchitecture, and after fixing with glutaraldhyde, the polyphenolic secretion was shown to remain as a single unit or was organized into round droplets. Transmission Electron Microscopy displayed immature plastids from young mesophyll cells containing eletron-dense deposits similar to phenolic substances, suggesting that Theobroma plastids are involved in the synthesis of phenolics.

Anatomy, ultrastructure and secretion of Hibiscus pernambucensis Arruda (Malvaceae) extrafloral nectary

This paper reports on the extrafloral nectary (EFN) of Hibiscus pernambucensis, a native shrub species occurring in mangrove and restinga along Brazil's coastline. EFNs occur as furrows with a protuberant border on the abaxial surface veins of the leaf blade. Each nectary consists of numerous secretory multicellular trichomes, epidermal cells in palisade-like arrangements and non-vascularized parenchyma tissue. Nectar secretion is prolonged, since secretion starts in very young leaves and remains up to completely expanded leaves. Reduced sugars, lipids, and proteins were histochemically detected in all the nectary cells; phenolic substances were detected in the vacuoles of the epidermal palisade cells and in some secretory trichome cells. The secretory cells that constitute the body of trichomes have large nuclei, dense cytoplasm with numerous mitochondria, dictyosomes, scattered lipid droplets and plastids with different inclusions: protein, lipid droplets or starch grains; vacuoles with different sizes have membranous material, phenolic and lipophilic substances. The palisade cells show thick periclinal walls, reduced cytoplasm with voluminous lipid drops and developed vacuoles. The nectary parenchyma cells contain abundant plasmodesmata and cytoplasm with scattered lipid droplets, mitochondria, plastids with starch grains and endoplasmic reticulum. Mucilage idioblasts are common in the inner nectary parenchyma. Protoderm and ground meristem participate in the formation of EFN. Our data indicate that all nectary regions are involved in nectar production and secretion, constituting a functional unit. Longevity of the extrafloral nectaries is likely associated with the presence of mucilage idioblasts, which increases the capacity of the nectary parenchyma to store water.

Leaf and calycine colleters in Odontadenia lutea (Apocynaceae - Apocynoideae - Odontadenieae): their structure and histochemistry

The structure and histochemistry of colleters found on the vegetative and floral apices of Odontadenia lutea are described. Colleters occur on vegetative apices starting at the fourth node, with 68 to 80 colleters being found at each node. Each leaf primordium has only one colleter of axillary origin, 3-5 intra-petiolar, and 12-16 inter-petiolar (intra-stipular). There are four types of colleters: standard, bipartite standard, sessile, and bipartite sessile. Colleters on the reproductive apices alternate with the sepals and are sessile, reduced sessile, tripartite laminar sessile, or asymmetrical. All of the colleters have a central nucleus of parenchymatous cells covered by a palisade uniseriate secretory epidermis and a thin cuticle. Secretory idioblasts were observed in the parenchymatous axis. Vascularization was observed only in standard axillary and laminar colleters. Crystals were observed in the parenchyma of the axillary colleter. Histochemical tests demonstrated that there was no rupturing or distension of the cuticle during the secretion process. Mucilage was identified using the PAS reaction as well as by Mayer's reagent and Ruthenium red staining. The calycine colleters had two distinct secretory phases, the first synthesizing mucilage and the second producing phenolic compounds.

Análise botânica, química e biológica comparativa entre flores das espécies sambucus nigra L. e sambucus australis cham. & Schltdt. e avaliação preliminar da estabilidade

Flores das espécies Sambucus nigra, de origem européia, e Sambucus australis, nativa da América do Sul (Caprifoliaceae), denominadas sabugueiro e sabugueirodo- brasil, respectivamente, são utilizadas popularmente sob forma de infusão, como antiinflamatórias, laxativas e para condições febris resultantes de afecções do trato respiratório. Estudos prévios para S. nigra indicam compostos fenólicos como principais constituintes químicos, sendo estes relacionados às principais atividades biológicas avaliadas. Objetivando comparar esta espécie com Sambucus australis, foram realizadas análises botânicas macro e microscópicas das flores, identificando as principais diferenças entre as espécies, tais como o número de lóculos no ovário e presença de idioblastos cristalíferos em algumas estruturas, e observando os possíveis contaminantes (pedicelos). Também foram determinados os parâmetros farmacopéicos: cinzas totais e perda por dessecação. Após a análise química, foi escolhido o flavonóide rutina como marcador das espécies, para realizar análises quantitativas nas 31 amostras adquiridas e / ou coletadas, utilizando método de CLAE previamente validado. Soluções hidroetanólicas apresentaram maior capacidade de extração do produto alvo. Os limites mínimos de rutina observados para ambas as espécies foram de aproximadamente 0,65%. Também foram quantificados, por método espectrofotométrico, os flavonóides totais expressos em quercetina, sendo 0,93% e 1,46% os teores mínimos determinados para S.nigra e S. australis, respectivamente. O estudo da estabilidade acelerada (50°C ± 90% U.R.), avaliando a degradação dos constituintes químicos presentes permitiu sugerir a cinética de degradação de segunda ordem para da rutina nas duas espécies. Comparações de atividades biológicas das espécies foram realizadas pelos ensaios das atividades antiinflamatória (inibição do edema em pata de rato induzido por carragenina) e antioxidante (DPPH). Os resultados para o primeiro ensaio demonstraram ação equivalente em ambas as espécies para extratos hidroetanólicos a 80% (86% de inibição) e aquosos (81%), com atividade semelhante ao padrão indometacina (~83%); para a atividade antioxidante os extratos hidroetanólicos a 80% foram mais ativos (CE50 = 16 μg/ml) que os aquosos (CE50 = 27 μg/ml) em S. australis, e ambos extratos, superiores ao 28 extrato padronizado Gingko biloba (CE50 = 40 μg/ml) e aos extratos de S. nigra (CE50= 50 μg/ml – hidroetanólico e CE50= 32 μg/ml – aquoso).

Análises morfológicas e moleculares dos gêneros Galium L.e Relbunium (Endl.)Hook.f. (RUbieae-Rubiaceae)no estado do Rio Grande do Sul-Brasil

Desde o início da história taxonômica de Relbunium, muitos foram os trabalhos que enfatizaram sua autonomia e posição taxonômica. Atualmente, alguns estudos sugerem que as espécies pertencentes a Relbunium devam ser incluídas em uma seção do gênero Galium. Porém, recentes estudos moleculares na tribo Rubieae, destacam Galium como um grupo parafilético, e Relbunium como um gênero independente e monofilético. O problema taxonômico referente a Galium e Relbunium é de difícil solução, devido à ausência de estudos que integrem caracteres morfológicos, ecológicos e moleculares. No presente trabalho objetivou-se adicionar informações para o conhecimento básico das espécies de Relbunium e Galium para o sul do Brasil, a partir de caracteres morfológicos e moleculares, buscando responder a seguinte questão: “Relbunium pode ser considerado um gênero ou apenas uma seção dentro de Galium?”. Para atingir os objetivos, foi analisada a morfologia das espécies, com ênfase nas folhas, flores e frutos para duas espécies de Galium e treze de Relbunium: G. latoramosum, G. uruguayense, R. equisetoides, R. gracillimum, R. hirtum, R. humile, R. humilioides, R. hypocarpium, R. longipedunculatum, R. mazocarpum, R. megapotamicum, R. nigro-ramosum, R. ostenianum, R. richardianum e R. valantioides. Chaves de identificação foram geradas a partir dos resultados das análises morfológicas. As folhas foram analisadas quanto à forma, ápice, padrão de venação, tricomas, estômatos, distribuição de idioblastos secretores e vascularização do hidatódio. Esses caracteres não evidenciaram a separação entre os gêneros, auxiliando apenas na individualização das espécies. A morfologia das flores e frutos auxiliou na diferenciação dos gêneros e espécies estudadas. As flores são comumente bispóricas, a exceção de G. latoramosum. Brácteas involucrais, ausentes em Galium, estão presentes nas espécies de Relbunium, de duas a quatro; nesse gênero há presença de antopódio, ausente em Galium. A corola possui tricomas glandulares unicelulares na face adaxial, e na face abaxial os tricomas, quando presentes, são simples e idioblastos secretores estão presentes apenas em R. gracillimum. O androceu tem quatro estames alternipétalos e exsertos, com anteras dorsifixas e tetrasporangiadas, de deiscência longitudinal. O ovário é ínfero, bicarpelar, bilocular, com um rudimento seminal anátropo e unitegumentado por lóculo. O desenvolvimento dos frutos, a estrutura do pericarpo e da testa foram descritos. Os frutos são do tipo baga, em R. gracillimum e R. hypocarpium, ou esquizocarpo, nas demais espécies. A consistência do pericarpo pode variar de carnosa, nos frutos do tipo baga, a levemente seca, nos frutos esquizocarpos. Entre as espécies, observou-se uma variação com relação ao exocarpo, que pode ser liso, piloso ou com idioblastos secretores. A testa é constituída por apenas uma camada de células, que em R. hypocarpium mostra-se descontínua. Além das descrições morfológicas, foram realizados estudos moleculares das espécies, através do seqüenciamento de fragmentos do DNA nuclear (ITS) e plastidial (trnL-F). A partir dos resultados obtidos formam elaborados cladogramas com base nos dados morfológicos e moleculares. O cladograma construído a partir dos dados morfológicos (vegetativos e reprodutivos) evidenciou a distinção dos dois gêneros, ou seja, sustenta Relbunium como táxon independente. Nesse cladograma observa-se que a VI presença ou ausência de brácteas foi determinante, e proporcionou a separação dos gêneros. A uniformidade dos caracteres morfológicos vegetativos entre as espécies auxiliou apenas na distinção das espécies de Relbunium. Com relação aos dados moleculares, os fragmentos de DNA utilizados mostraram-se pouco informativos. A análise do fragmento ITS, em especial, contribuiu para confirmação da relação entre algumas espécies (R. hirtum e R. ostenianum, e R. humile e R. mazocarpum). A análise combinada dos dados morfológicos e moleculares não caracterizou Relbunium como um clado monofilético, sendo sua manutenção não sustentada, isso, principalmente, devido à falta de diferenças moleculares entre as espécies. Conclui-se que para o grupo em questão as análises morfológicas, das folhas, flores e frutos, foram suficientes para destacar Relbunium como um gênero autônomo e monofilético na tribo Rubieae.

Phenolic compounds in leaves of Alchornea triplinervia: anatomical localization, mutagenicity, and antibacterial activity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização anatômica e fitoquímica de folhas e rizomas de Hedychium coronarium J. König (Zingiberaceae)

Devido a grande potencialidade na utilização de Hedychium coronarium, na medicina popular e também como biorremediadora no tratamento de efluentes, objetivou-se uma diagnose dos órgãos, folha e rizoma, para elucidar resultados estruturais e fitoquímicos. A folha é anfiestomática, com predominância de estômatos na face abaxial. em ambas as superfícies foliares há projeções de cera epicuticular sobre as paredes anticlinais das células epidérmicas. O mesofilo dorsiventral apresenta hipoderme multisseriada (3 camadas) em ambos os lados. O parênquima clorofiliano é diferenciado em paliçádico (1-2 camadas) e lacunoso (4-5 camadas) com muitos espaços intercelulares e ocorrência de idioblastos cristalíferos. Na nervura central, o aerênquima ocorre em único arco na região abaxial. Os feixes vasculares distribuem-se aleatoriamente e são de diferentes tamanhos, pequenos, médios e grandes, envolvidos por fibras. Os feixes menores localizam-se no lado abaxial da nervura. A triagem fitoquímica das folhas mostrou a presença de saponinas e ausência de taninos, antraquinonas, alcalóides e flavonóides. Por meio de Cromatografia em Camada Delgada foram identificadas as presenças de cariofileno e mirceno no óleo essencial bruto obtido a partir das folhas de H. coronarium.

Estruturas secretoras de mucilagem em Hibiscus pernambucensis Arruda (Malvaceae): distribuição, caracterização morfoanatômica e histoquímica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pulvinus functional traits in relation to leaf movements: A light and transmission electron microscopy study of the vascular system

Previous studies on legume pulvini suggest that the vascular system plays an important role in the redistribution of ions and transmission of stimuli during leaf's movements. However, the number of anatomical and ultrastructural studies is limited to few species. The aim of this paper is to investigate the structure and cellular features of the pulvinus vascular system of nine legume species from Brazilian cerrado, looking for structural traits pointing to its participation in the leaf's movements. Samples were excised from the medial region of opened pulvinus of Bauhinia rufa, Copaifera langsdorffii, Senna rugosa (Caesalpinioideae), Andira humilis, Dalbergia miscolobium, Zornia dilphylla (Faboideae), Mimosa rixosa, Mimosa flexuosa and Stryphnodendron polyphyllum (Mimosoideae), and were prepared following light microscopy, transmission electron microscopy and histochemical standard techniques. The vascular system occupies a central position, comprises phloem and xylem and is delimited by a living sheath of septate fibers in all the species studied. This living cells sheath connects the cortex to the vascular tissues via numerous plasmodesmata. The absence of fibers and sclereids, the presence of phenolic idioblasts and the abundance and diversity of protein inclusions in the sieve tube members are remarkable features of the phloem. Pitted vessel elements, parenchyma cells with abundant cytoplasm and living fibriform elements characterize the xylem. The lack of lignified tissues and extensive symplastic continuity by plasmodesmata are remarkable features of the vascular system of pulvini of the all studied species. (c) 2007 Elsevier Ltd. All rights reserved.