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Primer Archivo.Pdf Facultad de Ingenieria y Ciencias Acta de Correcciones al Proyecto de Grado Biología Fecha: 25 octubre 2020 Autores: Alfonso Andrade Campuzano Nombre del Proyecto de Grado: APLICACIÓN TÉCNICA DEL CÓDIGO DE BARRAS DE LA VIDA PARA EVALUAR LA DIVERSIDAD DE ESPECIES EN LA FAMILIA CRAMBIDAE (LEPIDOPTERA) EN LA RESERVA GUADUALITO, QUINDÍO. Directora: Nicola S. Flanagan Como indica el artículo 2.27 de las Directrices de Trabajo de Grado, he verificado que los estudiantes indicados arriba han implementado todas las correcciones que los Jurados del Proyecto de Grado definieron que se efectuaran, como consta en el Acta de Calificación correspondiente. ________________________________________ Firma del Directora del Proyecto de Grado Calle 18 No. 118-250 Av. Cañasgordas • A.A. Nos 8264 - 26239 • PBX 321 8200 • FAX 555 2180 - 555 2178 • www.puj.edu.co APLICACIÓN DE LA TÉCNICA DEL CÓDIGO DE BARRAS DE LA VIDA PARA EVALUAR LA DIVERSIDAD DE ESPECIES EN LA FAMILIA CRAMBIDAE (LEPIDOPTERA) EN LA RESERVA GUADUALITO, QUINDÍO. ALFONSO ANDRADE CAMPUZANO PONTIFICIA UNIVERSIDAD JAVERIANA CALI FACULTAD DE INGENIERÍA Y CIENCIAS DEPARTAMENTO DE CIENCIAS NATURALES Y MATEMÁTICAS CARRERA DE BIOLOGÍA SANTIAGO DE CALI 2020 APLICACIÓN DE LA TÉCNICA DEL CÓDIGO DE BARRAS DE LA VIDA PARA EVALUAR LA DIVERSIDAD DE ESPECIES EN LA FAMILIA CRAMBIDAE (LEPIDOPTERA) EN LA RESERVA GUADUALITO, QUINDÍO. ALFONSO ANDRADE CAMPUZANO DIRECTORA NICOLA S. FLANAGAN CODIRECTORA LUISA ARCILA - CARDONA PONTIFICIA UNIVERSIDAD JAVERIANA CALI FACULTAD DE INGENIERÍA Y CIENCIAS DEPARTAMENTO DE CIENCIAS NATURALES Y MATEMÁTICAS CARRERA DE BIOLOGÍA SANTIAGO DE CALI 2020 Resumen: La pérdida de biodiversidad a nivel global es un gran reto en términos ambientales, e identificar esta biodiversidad para implementar estrategias de conservación eficaces es el desafío de la taxonomía moderna. El método de Código de Barras de la Vida (Barcoding), es una aproximación desarrollada para identificar especies de una manera rápida y precisa usando regiones genéticas cortas y estandarizadas. Los lepidópteros son un orden altamente diverso que presenta varios casos de cripticismo entre sus especies. La familia Crambidae es una familia de polillas altamente diversa y difícil de identificar morfológicamente, y frecuentemente son confundidas con especies de su familia hermana Pyralidae. Las especies de la familia Crambidae han sido estudiadas por ser plagas de cultivos, pero se presenta un gran vacío en cuanto a la riqueza de esta familia. En este estudio se realizó un inventario de la diversidad de las polillas Crambidae en la Reserva Guadualito, ubicada en la zona Andina colombiana. Se muestrearon 45 individuos, de los cuales 24 correspondieron a 20 especies distintas con un porcentaje de identidad genética mayor a 97%. Las otras 21 muestras se pudieron identificar sólo hasta el nivel de género o taxonomía superior. De las 20 especies identificadas, 10 son nuevos registros para Colombia. Estos resultados confirman que el código de barras de la vida es una herramienta que aporta en el reconocimiento de la riqueza de especies de Lepidoptera, y se recalca que debe usarse más ampliamente para aumentar los registros de especies. Sin embargo, es esencial aplicar esta técnica de forma complementaria con la taxonomía basada en caracteres morfológicos, ya que las bases de datos de referencia (BOLD Systems y GenBank) todavía carece de un registro completo. Palabras Clave: Barcoding, Polillas, Molecular Operational Taxonomic Unit (MOTU), Taxonomía. Summary: The loss of biodiversity at a global level is a great challenge in environmental terms, and identifying this biodiversity to implement effective conservation strategies is the challenge of modern taxonomy. The Barcode of Life (Barcoding) method is an approach developed to identify species in a fast and precise way using short and standardized genetic regions. Lepidoptera are a highly diverse order that presents several cases of crypticism among its species. The Crambidae family is a highly diverse family of moths that is difficult to identify morphologically, and they are frequently confused with species from their sister family Pyralidae. Species of the Crambidae family have been studied for being crop pests, but there is a great gap in the richness of this family. In this study, an inventory of the diversity of Crambidae moths was carried out in the Guadualito Reserve, located in the Colombian Andean zone. 45 individuals were sampled, of which 24 corresponded to 20 different species with a percentage of genetic identity greater than 97%. The other 21 samples could be identified only up to the genus or higher taxonomy level. Of the 20 species identified, 10 are new records for Colombia. These results confirm that the life barcode is a tool that contributes to the recognition of the species richness of Lepidoptera, and it is emphasized that it should be used more widely to increase species records. However, it is essential to apply this technique in a complementary way with the taxonomy based on morphological characters, since the reference databases (BOLD Systems and GenBank) still lack a complete registry. Keywords: Barcoding, Moth, Molecular Operational Taxonomic Unit (MOTU), Taxonomy. Agradecimientos Quiero agradecerle en primer lugar a las profesoras Nicola S. Flanagan y Luisa Arcila por su entrega y acompañamiento incondicional durante la elaboración de este estudio. Al profesor Rodrigo Bernal, por permitirnos trabajar en la reserva Guadualito y acompañarnos fielmente en los muestreos con su gran conocimiento sobre la familia Crambidae. A mis compañeros/as de la carrera por estar siempre ahí dándome alientos, a mis compañeros/as de los laboratorios 2 y 3 por estar conmigo viviendo esta aventura ayudándonos mutuamente. Por último, a mi familia que siempre me ha apoyado constantemente. Tabla de contenido Introducción ................................................................................................................................... 1 Materiales y Métodos ................................................................................................................... 4 Área de estudio ....................................................................................................................... 4 Muestreo ................................................................................................................................... 4 Identificación Taxonómica ................................................................................................... 6 Generación de secuencias de ADN ................................................................................... 6 Análisis de la información ................................................................................................... 6 Generación del registro BOLD ............................................................................................ 7 Resultados ..................................................................................................................................... 7 Consulta en bases de datos Genbank y BOLD .............................................................. 7 Análisis de MOTUs ............................................................................................................... 14 Colectas repetidas del mismo MOTU .............................................................................. 14 Discusión ..................................................................................................................................... 15 Utilidad del código de Barras de la Vida ........................................................................ 15 Limitación en la información disponible en las bases de datos ............................. 16 Accesiones poco confiables ............................................................................................. 18 Futuros estudios .................................................................................................................. 19 Conclusión ................................................................................................................................... 20 REFERENCIAS BIBLIOGRÁFICAS ....................................................................................... 20 Introducción Colombia es uno de los países con mayor diversidad en el mundo (Poveda et al., 2010). Cuenta con un registro aproximado de 51 330 especies, cifra que lo posiciona como el país más biodiverso en aves y orquídeas, y el segundo en plantas, anfibios, peces dulceacuícolas y lepidópteros. En Lepidoptera son 1905 especies registradas hasta ahora, de las cuales aproximadamente el 16% (305) son endémicas (SiB Colombia, 2020). Esta biodiversidad se ve reflejada en el reconocimiento de dos importantes “hotspots” en el país: el Choco biogeográfico y los Andes tropicales, siendo este último, el hotspot más crítico para la conservación de la biodiversidad en la tierra (Myers et al., 2000). La gran biodiversidad de estos hotspots y en general del país se está perdiendo a causa de la transformación y deforestación de bosques naturales, ocasionadas por actividades como la ganadería extensiva, la agricultura y la minería (Poveda et al., 2010). Para tomar acciones que salvaguarden esta diversidad a nivel global, distintas entidades se han enfocado en generar un inventario de diversidad biológica (SCBD, 2020). El número de registros de todas las especies reconocidas en el planeta es aproximadamente de 1,8 a 1.9 millones y se estima que faltan entre 5 y 10 millones
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