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Andrade-Campuzano Articulo.Pdf 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 1, Luisa Arcila-Cardona 2, Nicola S. Flanagan 3 y Rodrigo Bernal 4. 1. Pontificia Universidad Javeriana-Cali. Correo: [email protected] 2. Universidad del Valle. Correo: [email protected] 3. Pontificia Universidad Javeriana-Cali. Correo: [email protected] 4. Reserva Natural Guadualito. Correo: [email protected] 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. 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 de especies por identificar (May y Harvey, 2009; Roskov et al., 2020). A pesar de los esfuerzos taxonómicos, la brecha de especies por describir, registrar e inventariar, se debe a la poca exploración de ciertas zonas del planeta y a los retos que representa la identificación morfológica de algunos taxa (Paz et al., 2011). Este problema es más difícil en grupos como insectos y plantas, debido a la gran diversidad que existe, combinado con la deficiencia en especialistas taxonómicos (Blaxter, 2003). El orden Lepidoptera es entre los insectos uno de los más numerosos, con aproximadamente 150.000 especies descritas (Van Nieukerken et al., 2011). La familia Crambidae, junto a Pyralidae conforman la super familia Pyraloidea, con alrededor de 16.000 especies en el mundo. Crambidae es la tercera familia con mayor número de especies dentro del orden (Solis, 2007) y la de mayor número de especies dentro de la super familia Pyraloidea (Acosta-Vásquez et al., 2017), con un total de 9,655 especies descritas en todo el mundo. Las especies de Pyraloidea se caracterizan morfológicamente por tener escamas en la base de la probóscide, órganos timpánicos abdominales ubicados en el esternito 2, y venas Rs2 y Rs3 del ala anterior fusionadas en una misma vena principal (Minet, 1991; Regier et al., 2012; Solis y Metz, 2016; Acosta-Vásquez, et al., 2017). La característica más relevante que diferencia a Crambidae de Pyralidae es la forma de los órganos timpánicos. En Pyralidae la cámara timpánica generalmente está cerrada o en algunos casos presenta una pequeña apertura anterior. En Crambidae, la cámara timpánica tiene una amplia apertura anteromedial y presenta una membrana en forma de colgajo llamada praecinctorium, ausente en Pyralidae (Guenée, 1854; Kristensen, 1998). Debido al tamaño pequeño de las especies, la diferenciación de ambas familias se basa en la observación detallada bajo el estereoscopio de estos órganos timpánicos (Solis, 2007). A simple vista Crambidae y Pyralidae son indistinguibles, a menos que sean observadas por un taxónomo experto (Regier et al., 2012). De igual forma, al interior de ambas familias, la taxonomía se basa en rasgos de genitalia inconspicuos. Estas características de difícil 1 observación, sumado a su gran diversidad, ocasiona que las especies crípticas de este grupo taxonómico sean obviadas comúnmente (Smith et al., 2006). Por ende, a pesar de ser una de las familias más numerosas entre los lepidópteros (Solis, 2007), la taxonomía de la familia Crambidae siempre ha tenido brechas y muy pocos avances sistemáticos en comparación a otras familias de lepidópteros (Regier et al., 2012) La familia Crambidae incluye especies que son plagas importantes, que causan daños económicos a los cultivos, bosques y productos almacenados (Nagaharish et al., 2017; Solis, 2007; Solis y Metz, 2016). Por ejemplo, en Colombia recientemente se ha descrito la nueva especie Eoreuma insuastii como plaga importante de la caña de azúcar (Solis et al., 2020a). A pesar de la importancia económica de este grupo, su taxonomía en Colombia no se ha estudiado de manera exhaustiva, debido a la dificultad para encontrar caracteres diagnósticos que permitan separar especies en esta familia con el uso de la taxonomía morfológica. La identificación taxonómica de algunas especies representa grandes desafíos cuando se utilizan métodos enfocados en la delimitación a partir de rasgos morfológicos diferenciales. Los retos de clasificación en estos casos se originan, ya sea porque la especiación ocurre sin cambios morfológicos o porque las estructuras morfológicas son difíciles de estudiar (Padial et al., 2010). Ambos casos pueden resultar en grupos de especies crípticas, las cuales se definen como dos o más especies clasificadas como una sola especie nominal porque morfológicamente son indistinguibles (Bickford et al., 2007). En los lepidópteros la ocurrencia de especies crípticas se ha detectado a través de diferentes familias, como Hesperiidae (Hebert et al., 2004), Papilionidae (Lukhtanov et al., 2016), Pyralidae y Crambidae (Haines y Rubinoff, 2012; Regier et al., 2012). El reconocimiento de esta diversidad
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