Anatomía microscópica y metabolitos secundarios volátiles en tres estadios del desarrollo de las inflorescencias de Lantana camara (Verbenaceae)

Microscopic anatomy and volatile secondary metabolites at three stages of development of the inflorescences of Lantana camara (Verbenaceae). Plants of the Verbenaceae family, like L. camara, have called the attention of researchers, not only because of its high diversity and its distribution around the world, but also for its variable use as popular medicine to treat diseases like tetanus, rheumatism and malaria, and as bactericide and insecticide. To assess this, the morphology and ontogeny of the inflorescences of Lantana camara and the chemical composition of volatile secondary metabolites were analyzed at three different ontogeny stages. Plants were collected from the experimental crop area in CENIVAM, Bucaramanga, Colombia. Fresh inflorescence stages were established and analyzed using a stereoscopic microscope, fixed in FAA and included in parafine. Transversal and longitudinal 10μm thick sections were prepared using a rotative microtome, safranine-fastgreen stained and were observed and photographed using a light microscope. The chemical composition of volatile secondary metabolites were analyzed for each stage. The analytes, obtained from 0.7g of plant, were isolated by solid phase micro-extraction in the headspace mode (HS-SPME) and were placed in 20ml vials. The components were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Stage I was microscopically characterized by an immature development in which the meristematic differentiation begins with a mass of cells. In Stage II, the morphogenetic movement gives way to the formation of the respective floral sexual structures, calyx and corolla. In Stage III, the different organs are conspicuous: four stamens epipetals and didynamous, monocarpelar, biloculate and globose gynoecium, upper ovary and lateral stigma; the flowers are hermaphroditic. The main secondary metabolites detected by GC-MS were bicyclosesquiphellandrene, E-β-farnesene, E-β-caryophyllene, Υ-muurolene + Υ- curcumene and α-zingiberene. Nevertheless, this study reports for the first time in plant species α-gurjunene, Υ-amorphene, α-muurolene, sesquithujene, α-trans-bergamotene and transcadina-1,4-diene. The diversity of compounds found can be only explained by the extraction methods employed, the developmental stages and section of the plant, the geographic conditions, collection time and the genetic constitution of the evaluated species. Rev. Biol. Trop. 59 (1): 473-486. Epub 2011 March 01.

Las plantas de la familia Verbenaceae, entre ellas la especie Lantana camara L., poseen una alta diversidad botánica además de su variable uso. Se describe el avance morfoanatómico de tres etapas de desarrollo de las inflorescencias de L. camara L. (Verbenaceae), recolectadas en Bucaramanga, Colombia. Para esto se tincionaron las estructuras vegetales mediante la técnica fast-green, previo a los análisis con microscopía óptica convencional. El aislamiento de los analitos de cada estadio se realizó mediante la técnica de microextracción en fase sólida en el modo headspace (HS-SPME) y su posterior análisis de componentes, mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS). En el Estadio I se apreció un grupo de células en donde se inicia la diferenciación meristemática. En el Estadio II, los movimientos morfogenéticos dan paso a la formación de las respectivas estructuras sexuales florales, el cáliz y la corola. Al llegar al Estadio III, se observaron claramente los diferentes órganos. Los metabolitos detectados en este estudio y que no fueron encontrados en la bibliografía revisada fueron, el α- gurjuneno, el Υ-amorfeno, el α-muuroleno, el sesquitujeno, el α-trans- bergamoteno y el trans-cadina-1,4-dieno. Los principales metabolitos secundarios detectados fueron: biciclosesquifelandreno, E-β-farneseno, E-β-cariofileno, Υ- muuroleno+Υ-curcumeno y α-zingibereno. Una función importante de éstas y otras esencias florales es la defensa y la protección durante el estrés abiótico de la planta.

Morfología, anatomía, ontogenia y composición química de metabolitos secundarios en inflorescencias de Lippia alba (Verbenaceae)

Morphology, anatomy, ontogeny and chemical composition of inflorescences volatile secondary metabolites of Lippia alba (Verbenaceae) at three stages of development. There is an increased interest to know and scientifically validate traditional knowledge of medicinal plants. Lippia alba belongs to Verbenaceae family and has been of interest, not only because of its worldwide extensive distribution, but also for its variable use as antiviral, bactericide, citostatic, analgesic and sedative. To study this, the morphology and ontogeny of Lippia alba inflorescences and the chemical composition of its volatile secondary metabolites were analyzed during three different stages of development. Plants were collected at the experimental crop field in CENIVAM, Bucaramanga, Colombia. The inflorescence’s morphology and ontogeny, and the chemical composition of volatile secondary metabolites were analyzed using a stereoscopic microscope and chromatographic and spectroscopic techniques. Fresh material corresponding to each stage was fixed in F.A.A (formol, acetic acid and alcohol), included in paraffin and cutted in transversal and longitudinal sections. Sections were stained with safranine-fastgreen, photographed and decribed. The chemical composition of volatile secondary metabolites at each ontogenic stage, was extracted by solid phase micro-extraction in the headspace mode and analyzed by gas chromatography coupled to mass spectrometry. Stage I showed a meristematic mass of cells in vegetative apex and bracts, with an outline of floral whorls. In Stage III, the stamens were adnate, epipetals and didynamous, bicarpelar and syncarpic gynoecium, with superior ovary and decurrent stigma. The main secondary metabolites detected were the bicyclosesquiphellandrene followed by carvone, limonene and trans-β-farnesene, that constituted the 78% of the total relative amounts of compounds. Other metabolites such as β-copaene, γ-amorphene and cis-β-guaiene, were reported for the first time in this study. When compared to other studies, morphological differences reported in this study are possibly related to adaptation to environmental conditions or pollinators, which let us suggest that there is no specific ontogenic pattern. Similarly, the qualitative and quantitative variations in the detected compounds could be explained because one or more of them are used as precursors of others. Rev. Biol. Trop. 58 (4): 1533-1548. Epub 2010 December 01.

La especie Lippia alba (Verbenaceae) llama la atención de los investigadores por su amplia distribución en todo el mundo y sus diversas propiedades. Se establecieron tres etapas durante el desarrollo de sus inflorescencias, fueron recolectadas en Bucaramanga, Colombia para analizar los cambios morfoanatómicos. Para ello, las muestras se trataron mediante la técnica safraninafastgreen y se observaron utilizando microscopio óptico convencional. Se realizó el aislamiento de los metabolitos secundarios volátiles usando la técnica de microextracción en fase sólida en el modo headspace (HS-SPME) y su posterior análisis de componentes, mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS). En la Etapa I se observó una masa de células meristemáticas en el ápice vegetativo. En la Etapa II la acción de procesos morfogenéticos dan paso a la formación de los verticilos florales del perianto. En la Etapa III se establecen todos los verticilos florales. Los principales metabolitos secundarios constituyen el 78% de la cantidad relativa total. Ellos son el biciclosesquifelandreno, la carvona, el limoneno y el trans-β-farneseno. Los metabolitos β-copaeno, γ-amorfeno y cis-β-guaieno, no estαn registrados en la bibliografνa revisada. Estos compuestos activos pueden variar de acuerdo con el estado de desarrollo de la planta, la parte empleada para la extracciσn y el mιtodo implementado para ello.