Resumen de Caracterización agronómica y genómica de selecciones obtenidas en un programa de mejora de olivo para resistencia a la verticilosis
1. Introducción o motivación de la tesis En las últimas décadas la olivicultura ha experimentado grandes cambios dirigidos a incrementar la rentabilidad y a reducir los costes de manejo del cultivo del olivo. Estos cambios han consistido principalmente en la intensificación de los olivares tradicionales y en el aumento de la superficie cultivada en todo el mundo. Además, los avances científicos sobre las propiedades saludables de los compuestos del aceite de oliva han desencadenado un aumento de la demanda mundial y el interés por los aceites de oliva vírgenes extra (AOVEs) de alta calidad. No obstante, han surgido también ciertas limitaciones, como la aparición o propagación de enfermedades como la Verticilosis (VO) y la falta de variedades adaptadas a plantaciones intensivas o a una recolección mecanizada.
En base a lo anterior, una de las líneas de trabajo del programa de mejora de olivo desarrollado en el Centro 'Alameda del Obispo' del Instituto de Investigación y Formación Agraria y Pesquera de Andalucía (IFAPA) es la búsqueda de nuevas variedades resistentes a la VO, con características agronómicas mejoradas y adecuada calidad del aceite. En este sentido, fruto de trabajos realizados en etapas previas, el programa de mejora cuenta con 20 preselecciones potencialmente resistentes y altamente productivas que han pasado a evaluaciones a largo plazo en ensayos comparativos. Esta tesis incluye los trabajos realizados en campo con dichas preselecciones para confirmar su resistencia en condiciones naturales de cultivo y evaluar las características productivas y calidad de sus aceites. Por otra parte, también se ha incluido un estudio a nivel genético de un conjunto de genotipos de diferente procedencia y respuesta a la VO, para tratar de identificar polimorfismos en regiones de genes candidatos para la resistencia.
2. Contenido de la investigación El primer objetivo de esta tesis consistió en evaluar 20 preselecciones frente al patotipo defoliante de Verticillium dahliae en condiciones semicontroladas (Capítulo 2). Para esta evaluación se realizó un experimento en microparcelas, en el que se plantaron los genotipos seleccionados tanto en suelo libre de inóculo como en suelo infestado artificialmente con microesclerocios de V. dahliae. Los resultados mostraron discrepancias en comparación con la respuesta a la enfermedad obtenida en evaluaciones bajo condiciones controladas, ya que 11 genotipos resultaron susceptibles. Además, las plantas que crecieron sobre el suelo infestado mostraron una reducción del crecimiento y un retraso en la entrada en producción, independientemente del desarrollo de síntomas de la enfermedad.
Estos genotipos también fueron evaluados en ensayos comparativos instalados en fincas de agricultores distribuidas por la provincia de Jaén (Andalucía, España). Estas fincas estaban infestadas por V. dahliae, por lo que la resistencia a la enfermedad fue reevaluada en condiciones naturales de cultivo. Aunque los resultados de dicha evaluación no constan en esta, los datos registrados están siendo analizados para una futura publicación. Sin embargo, como resultado de ellos, se seleccionaron tres genotipos por su mayor resistencia a la enfermedad (IFAPA111-2, IFAPA117-117 e IFAPA117-120) y la variedad de referencia ‘Picual’ se seleccionaron para realizar extracción de aceite y analizar su composición.
Un primer análisis, incluido en el capítulo 3 de la tesis, consistió en evaluar la variabilidad de 66 compuestos químicos, la estabilidad oxidativa y los atributos sensoriales en aceites de 4 genotipos diferentes (las selecciones IFAPA111-2, IFAPA117-117 e IFAPA117-120 y la variedad de referencia ‘Picual’) cultivados en 4 localidades (Córdoba, Úbeda, Begíjar y Arjona). El análisis estadístico aplicado ha revelado la posibilidad de predecir de forma precisa la estabilidad del AOVE en función de su composición, influyendo positivamente el contenido de ácido oleico y 3,4-DHPEA-EA. Por otra parte, para los atributos sensoriales no se ha observado una clara correlación con la composición química del AOVE y, tampoco, un efecto significativo del genotipo o el ambiente de cultivo.
El segundo análisis se centró en determinar de forma más específica el efecto de los factores genético (G) y ambiental (A), así como la interacción de ambos (G x A), en la composición volátil de los AOVEs, un tipo de estudio que no se había realizado previamente en olivo y que ha sido descrito en el Capítulo 4 de la tesis doctoral. Los resultados revelaron que el factor genético influye principalmente en el perfil volátil y el factor ambiental afecta al contenido total, mientras que el efecto G x A sólo se ha observado en los compuestos C5/LA y ésteres. Atendiendo a la fracción volátil, se ha demostrado que las selecciones del programa de mejora podrían ser una alternativa para la diversificación de variedades y ampliar la oferta comercial de los AOVEs. Por último, en el Capítulo 5 se describe el estudio a nivel genético realizado para analizar las diferencias genéticas entre 77 genotipos con diferente nivel de resistencia a la enfermedad, incluyendo variedades del Banco Mundial de Germoplasma de Olivo de IFAPA, acebuches y subespecies afines (cerasiformis y guanchica) de la Colección de olivo silvestre y progenies del programa de mejora. Concretamente, se han estudiado polimorfismos en fragmentos de 7 genes asociados a diferentes mecanismos de respuesta a la Verticilosis. En total se han identificado 92 SNPs y 2 InDels. Los SNPs detectados en el gen TLP1 podrían ser de utilidad para distinguir entre subespecies y una delección en el gen PFN2 podría estar relacionada con la respuesta de resistencia. Estas diferencias nucleotídicas podrían contribuir al establecimiento de marcadores para la gestión de colecciones de germoplasma y la selección en programas de mejora genética de olivo.
3. Conclusiones El desarrollo de nuevas variedades de olivo es un proceso largo que requiere de ensayos comparativos en diferentes condiciones, ya que la respuesta a la enfermedad o las características agronómicas pueden verse afectadas por el ambiente de cultivo. En el caso de la evaluación de resistencia, se han observado discrepancias en la respuesta de algunos genotipos dependiendo de las condiciones de evaluación. En este sentido, algunas selecciones potencialmente resistentes en condiciones controladas en cámaras de cultivo han resultado susceptibles en condiciones semicontroladas en microparcelas. Al mismo tiempo, se ha observado que los efectos negativos de la Verticilosis, como el retraso en la entrada en producción y la reducción del crecimiento, son menos acentuados en genotipos resistentes, independientemente del desarrollo de síntomas de la enfermedad.
En cuando a la calidad de los aceites de las selecciones resistentes a la Verticilosis, se ha observado que la variabilidad en la composición de los aceites es una característica principalmente dependiente del factor genético para la mayoría de los compuestos, lo que indica que los programas de mejora pueden contribuir al aumento de diversidad en la oferta de los aceites de oliva vírgenes extra. Así, los aceites extraídos de la selección ‘IFAPA111-2’ podrían ser similares al de ‘Picual’ y claramente diferenciados de las selecciones ‘IFAPA117-117’ e ‘IFAPA117-120’. Así mismo, se ha observado que la estabilidad oxidativa de los aceites de oliva vírgenes extra (vida útil) podría ser predecible en función de la composición de estos, ejerciendo un efecto positivo el contenido en ácido oleico, escualeno y compuestos fenólicos. Mientras que las características sensoriales de frutado, amargo y picante, no han resultado dependientes de los compuestos analizados.
A pesar del efecto mayoritario del factor genético, el contenido en compuestos volátiles y fenólicos se ha visto principalmente influenciado por el ambiente. Además, se ha observado un efecto significativo de la interacción genotipo por ambiente sobre algunos compuestos del aceite de oliva virgen extra, lo que demuestra la necesidad de realizar ensayos comparativos en condiciones naturales de cultivo para caracterizar la calidad de los aceites de las selecciones del programa de mejora.
Los estudios a nivel genético han identificado varios polimorfismos en genes candidatos para la resistencia a la VO. Sin embargo, su implicación en la respuesta del olivo a la infección por el hongo V. dahliae debe ser validada en futuros estudios, ya que dicha respuesta es muy variable entre genotipos e influenciable por las condiciones ambientales.
Como resultado de todos estos trabajos de caracterización agronómica y de resistencia se ha solicitado el registro de tres nuevas variedades que serán adecuadas para su plantación en zonas donde haya riesgo de incidencia de Verticilosis y con unas características diferenciadas de sus aceites. Igualmente, el procedimiento llevado a cabo para la obtención de variedades resistentes a la Verticilosis podría aplicarse también para el desarrollo de variedades resistentes a otras enfermedades del olivo, como Xylella fastidiosa, realizando las evaluaciones específicas en cada etapa del programa de mejora de olivo.
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