Anatomía microscópica y metabolitos secundarios volátiles en tres estadios del desarrollo de las inflorescencias de Lantana camara (Verbenaceae)
José Fernando Caroprese Araque, María Isabel Parra Garcés, Dagoberto Arrieta Prieto & Elena Stashenko
ABSTRACT
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, γ-muurolene + γ-curcumene and α-zingiberene. Nevertheless, this study reports for the first time in plant species α-gurjunene, γ-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.
RESUMEN
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 γ-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, γ-muuroleno+γ-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.