Utilización de un set de microsatélites para la tipificación y determinación de su huella alélica en cultivares de olivo de la provincia de Jaén
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2018-03-16
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Jaén: Universidad de Jaén
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[ES] Olea europea L es uno de los cultivos más importantes en la cuenca Mediterránea, especialmente para la economía de península Ibérica, por lo que cualquier avance en investigación es de vital importancia para el futuro de este cultivo. En este trabajo se ha obtenido la huella alélica de cinco variedades de olivo oriundas de la provincia de Jaén mediante el uso de ocho marcadores microsatélites previamente diseñados y contrastados con 36 cultivares correspondientes a la colección nuclear de del Banco Mundial de Germoplasma de Córdoba. Para ello, se realizó un único muestreo de las variedades a identificar en la estación experimental de olivicultura IFAPA “Venta del Llano”, de Mengíbar (Jaén). Todas las variedades se recolectaron por duplicado eligiendo dos ejemplares de olivo distintos procedentes de dicho banco de germoplasma. Para llevar a cabo la tipificación se ha puesto a punto en este trabajo la metodología PCR múltiple, introduciendo cinco marcadores SSRs (single sequence repeat) del set mencionado anteriormente por cada PCR ensayada. Una vez procesadas las muestras, y amplificadas mediante PCR, los datos de secuenciación se procesaron con un secuenciador capilar de ADN 3500 Genetic Analycer (Life technologies), utilizando el estándar interno GeneScan ™ 600 LIZ® Tamaño Estándar v2.0 (Life technologies), y los resultados se analizaron con el programa informático GeneMarker V2.6.3., obteniendo finalmente la huella alélica para los ocho marcadores en cada una de las diez muestras analizadas.
[EN] Olea europea L is one of the most important crops in the Mediterranean basin, especially for the economy of the Iberian Peninsula, so any progress in research is of vital importance for the future of this crop. In this work the allele trace of five olive varieties originating in the province of Jaén has been obtained through the use of eight microsatellite markers designed and contrasted with 36 cultivars corresponding to the nuclear collection of the World Bank of Germplasm of Córdoba. For this purpose, a single sampling of the varieties to be identified was carried out at the IFAPA "Venta del Llano" olive-growing experimental station in Mengíbar (Jaén). All the varieties were collected by duplicate by choosing the different olive specimens from the germplasm bank. To carry out this work, a multiplex PCR type methodology was developed introducing five SSRs (single sequence repeat) markers from the set of eigth microsatellites, in the PCR tubes. After the samples were amplified, the sequencing data were processed with a 3500 Genetic Analyzer (Life technologies) DNA capillary sequencer using the GeneScan ™ 600 Standard LIZ® Standard Standard v2.0 (Life technologies). The results were analyzed with the GeneMarker V2.6.3 software, finally obtaining the allelic profiles for the eight markers in each of the ten samples analyzed.
[EN] Olea europea L is one of the most important crops in the Mediterranean basin, especially for the economy of the Iberian Peninsula, so any progress in research is of vital importance for the future of this crop. In this work the allele trace of five olive varieties originating in the province of Jaén has been obtained through the use of eight microsatellite markers designed and contrasted with 36 cultivars corresponding to the nuclear collection of the World Bank of Germplasm of Córdoba. For this purpose, a single sampling of the varieties to be identified was carried out at the IFAPA "Venta del Llano" olive-growing experimental station in Mengíbar (Jaén). All the varieties were collected by duplicate by choosing the different olive specimens from the germplasm bank. To carry out this work, a multiplex PCR type methodology was developed introducing five SSRs (single sequence repeat) markers from the set of eigth microsatellites, in the PCR tubes. After the samples were amplified, the sequencing data were processed with a 3500 Genetic Analyzer (Life technologies) DNA capillary sequencer using the GeneScan ™ 600 Standard LIZ® Standard Standard v2.0 (Life technologies). The results were analyzed with the GeneMarker V2.6.3 software, finally obtaining the allelic profiles for the eight markers in each of the ten samples analyzed.