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Análisis Morfológico Del Sistema Reproductor E Identificación

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Análisis Morfológico Del Sistema Reproductor E Identificación Revista Peruana de Biología ISSN: 1561-0837 [email protected] Universidad Nacional Mayor de San Marcos Perú Jaramillo Roldán, Erika; López Martínez, Jessika; Ramírez, Rina; Velásquez Trujillo, Luz Elena Análisis morfológico del sistema reproductor e identificación molecular a través de los marcadores mitocondriales COI y 16S rRNA de Megalobulimus oblongus (Mollusca, Strophocheilidae) de Colombia Revista Peruana de Biología, vol. 21, núm. 1, mayo-, 2014, pp. 79-88 Universidad Nacional Mayor de San Marcos Lima, Perú Disponible en: http://www.redalyc.org/articulo.oa?id=195031025004 Cómo citar el artículo Número completo Sistema de Información Científica Más información del artículo Red de Revistas Científicas de América Latina, el Caribe, España y Portugal Página de la revista en redalyc.org Proyecto académico sin fines de lucro, desarrollado bajo la iniciativa de acceso abierto Revista peruana de biología 21(1): 079 - 088 (2014) ISSN-L 1561-0837 Identificación morfológica y molecular de MEGALOBULIMUS OBLONGUS DE COLOMBIA doi: http://doi.org/10.15381/rpb.v21i1.8250 FACULTAD DE CIENCIAS BIOLÓGICAS UNMSM TRABAJOS ORIGINALES Análisis morfológico del sistema reproductor e identificación molecular a través de los marcadores mitocondriales COI y 16S rRNA de Megalobulimus oblongus (Mollusca, Strophocheilidae) de Colombia Morphological analysis of the reproductive system and molecular identification by mitochondrial markers COI and 16S rRNA of Megalobulimus oblongus (Mollusca, Strophocheilidae) of Colombia Erika Jaramillo Roldán1,3, Jessika López Martínez1,3, Rina Ramírez2, Luz Elena Velásquez Trujillo1,3 1 Grupo de Microbiología Ambiental, Escuela de Resumen Microbiología, Universidad de Antioquia, Medel- lín, Colombia. En este trabajo se hizo el análisis morfológico y molecular a 28 caracoles terrestres de Mega- 2 Laboratorio de Sistemática Molecular y Filogeo- lobulimus oblongus, colectados en diferentes departamentos de Colombia, depositados en grafía, Facultad de Ciencias Biológicas; y Museo una colección de referencia. Para la caracterización morfológica, los animales se disecaron de Historia Natural, Universidad Nacional Mayor al estéreomicroscopio. Se describieron el sistema reproductor y la concha. Del sistema re- de San Marcos, Av. Arenales 1256, Lima-11, Perú. productor se tomaron medidas a las estructuras que lo componen. De la concha se describió 3 Programa de Estudio y Control de Enfermedades la forma, el color, el número de espiras y la ornamentación e igualmente se tomaron medidas Tropicales – PECET, Sede de Investigación Uni- básicas usando un calibrador digital. Para el análisis molecular se usaron dos marcadores versitaria – SIU, Medellín, Colombia. mitocondriales, el 16S rRNA y el gen Citocromo Oxidasa C subunidad I (COI). Ambos mar- Autora para correspondencia: Rina Ramírez, cadores confirmaron la presencia de un único haplotipo en los ejemplares, aun para indi- Email: [email protected] viduos situados en regiones biogeográficas diferentes y distantes. Este estudio sugiere que Email Jessika López: [email protected] en Colombia M. oblongus se encuentra en peligro, por lo que urgen investigaciones sobre Email Erika Jaramillo: [email protected] reproducción, genética poblacional y biogeografía que esclarezcan su situación en el país. Email Luz Velásquez: [email protected] Demuestra además que las colecciones de referencia y los bancos de tejidos son fuentes de información de gran valor, ya que permiten conocer aspectos relacionados con el riesgo en que se encuentran las especies y que sirven de insumo para el diseño de acciones de conservación. Palabras claves: 16S rRNA; COI; caracterización morfológica; Megalobulimus oblongus; Colombia. Citación: Abstract Jaramillo Roldán E., J. López Martínez, R. In this work was done morphological and molecular analysis to 28 land snails of Megalobulimus Ramírez, L.E. Velásquez Trujillo. 2014. Análisis oblongus, collected in different departments of Colombia, deposited in a reference collection. morfológico del sistema reproductor e identifi- cación molecular a través de los marcadores For morphological characterization, the animals were dissected in a stereomicroscope. The mitocondriales COI y 16S rRNA de Megalobulimus reproductive system and the shell were described. Measures were taken to structures of oblongus (Mollusca, Strophocheilidae) de Colom- the reproductive system. Of the shell were described its shape, color, number of whorls and bia. Revista peruana de biología 21(1): 079- 088 ornamentation and equally basic measures were taken using a digital caliper. For molecular (Mayo 2014), doi: http://doi.org/10.15381/rpb. analysis were used two mitochondrial markers, 16S rRNA and cytochrome C oxidase subunit v21i1.8250 I (COI). Only one haplotype was obtained for each marker, even for individuals of different and distant biogeographical regions. This study suggests that M. oblongus is in danger, therefore are urgent investigations about reproduction, population genetics and biogeography to clarify its situation in Colombia. It also demonstrates that the reference collections and tissue banks are sources of valuable information since they allow knowing aspects related with the species´ risk that serve as an input for the design of conservation actions. Keywords: 16S rRNA; COI; morphology; Megalobulimus oblongus; Colombia. Presentado: 13/12/2013 Aceptado: 16/03/2014 Publicado online: 26/05/2014 Journal home page: http://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/index © Los autores. Este artículo es publicado por la Revista Peruana de Biología de la Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos. Este es un artículo de acceso abierto, distribuido bajo los términos de la Licencia Creative Commons Atribución-NoComercial-CompartirIgual 4.0 Internacional.(http://creativecommons.org/licenses/by-nc-sa/4.0/), que permite el uso no comercial, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citadas. Para uso comercial, por favor póngase en contacto con [email protected]. Rev. peru. biol. 21(1): 079- 088 (May 2014) 79 Jaramillo Roldán et al. Introducción Por lo tanto nuestra investigación se propuso establecer el es- La familia Strophocheilidae (Hausdorf & Bouchet 2005) (Mol- tatus taxonómico de los caracoles terrestres, identificados apriori lusca: Gastropoda: Stylommatophora) de origen gondwánico, como Megalobulimus oblongus (Müller 1774) y depositados en la incluye los moluscos terrestres del género Megalobulimus Miller colección de Moluscos Vectores de la Universidad de Antioquia, 1878, endémicos de la región Andina y Amazónica Suramericana utilizando la caracterización morfológica de las partes blandas (Leme 1975), considerados los más grandes del continente. del caracol, además de la identificación molecular con base en los marcadores mitocondriales 16S rRNA y COI. Las clasificaciones taxonómicas de las especies de Mega- lobulimus, realizadas en los siglos XVIII y XIX, con base en los Material y métodos caracteres morfológicos de la concha, describieron 50 especies, Material biológico.- Para esta investigación se analizaron 28 siete de las cuales fueron registradas para Colombia: M. popelair- ejemplares de Megalobulimus oblongus adultos, ya que las conchas ianus (Nyst 1845), M. thammianus (Martens 1876), M. senezi de todos tenían una altura total mayor de 90 mm y el labio (Jousseaume, 1887), M. oblongus (Müller 1774), M. auritus, externo doblado hacia afuera y rosado. Los especímenes forman M. valenciennesii (Pfeiffer 1842) y M. pudicus (Müller, 1774). parte de la colección de Moluscos Vectores de la Universidad La distribución geográfica en Colombia varía según la especie, de Antioquia VHET N°37 y provienen de seis localidades de por ejemplo M. thammianus se señala para la región andina, M. Colombia (Tabla 1). Las partes blandas de los caracoles estaban oblongus y M. popelairianus para las regiones Caribe, Andina, conservadas en Railliet-Henry, las conchas almacenadas en seco Amazonia y Orinoquia. En tanto que M. valenciennesii solo se y un trozo de tejido del pie conservado en alcohol al 96%. reporta para la amazonia; se desconoce la procedencia de M. Descripción morfológica del aparato reproductor y la auritus, M. pudicus y M. senezi (Daniel 1942, Bequaert 1948, concha de Megalobulimus.- Para la descripción del sistema Vera 2008, Restrepo 2009, Linares & Vera 2012). reproductor se disectaron los 28 ejemplares de M. oblongus La validez de las determinaciones taxonómicas específicas de bajo estéreo microscopio (Nikon SMZ 745T); a cada una de los Megalobulimus fue cuestionada por Bequaert (1948) quien las estructuras que lo componen se le midió el largo y el ancho, consideró que la conquiliología era insuficiente para permitir este con la reglilla ocular. A las conchas se les midió el largo y ancho diagnóstico. Desde entonces se busca incluir las descripciones de total y de la apertura, usando un calibrador digital (Uiustools® la anatomía interna, con énfasis en el sistema reproductor (Leme MT-0085cun, precisión de ±0.02 mm). Las dimensiones se 1973, 1989, 1993, Leme & Indrusiak 1990). Sin embargo, dan en milímetros. Se realizaron registros fotográficos (Cámara en Colombia no ha sido estudiada la morfología de las partes Sony de 7.2 MP y ZOOM 2,8-5,2/ 6,3- 18,9 mm) y el aparato blandas de los Megalobulimus mencionados. reproductor se ilustró en el software Corel Draw X3. Si bien los caracteres morfológicos dan una buena resolución Extracción y amplificación de
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