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Trabajo presentado a la II Jornada sobre Investigación Biomédica, celebrada en la Universidad de Jaén el 20 de diciembre de 2012.

Resumen del póster presentado al Plant Biology Congress, celebrado en Friburgo (Alemania) del 29 de julio al 3 de agosto de 2012., ROS and NO are low molecular weight molecules implicated in plant signalling. Their relevant implication in pollen tube growth and reorientation has been shown particularly in plant model species. In olive reproductive tissues, their presence has been described in a stage- and tissue- specific manner. Accurate detection and quantification of minor changes in ROS/NO and the chemical species resulting or interacting with their metabolism in reproductive tissues is a challenging task at present. The goal of the present study was to elucidate the implication of the NO metabolism in the process of olive pollen germination in vítro. For this purpose we used a recently published method based in the use of liquid chromatography-electrospray/mass spectrometry to detect different antioxidant metabolites in volved in NO metabolism. We have analysed the levels of key molecules like GSH (reduced glutathione), GSSH (oxidized glutathione), GSNO {Snitrosoglutathione) and ascorbic acid (vitamin e) in the mature pollen, and the pollen grain and pollen tube at different times of in vitro germination. These data correspond to both dormant and activated metabolism, respectively. The results revealed remarkalble changes in the levels of GSNO, which is a cellular NO reservoir, throughout the germination process. These data confirm the involvement of NO and antioxidant molecules in the pollen tube growth, as observed previously by using CLSM, and point to potential new specific roles for these molecules as signalling intermediates., Work supported by ERDF-cofounded project MCINN BFU2011-22779. MJJQ and AZ thanks JAE grant funding.

Resumen del póster presentado al IV Spanish and Portuguese Meeting on Free Radicals, celebrado en Valencia (España) del 5 al 7 de junio de 2012., Olive pollen is a major cause of allergy in Mediterranean countries. Numerous studies are devoted to the characterization of allergy-eliciting proteins from this allergenic source. However, recent research is focusing on the study of other non-allergenic protein and non-protein components modulating the allergic immune response. These molecules include pollen-associated lipid mediators, adenosine, proteases and oxidases, particularly NAD(P)H oxidase (NOX) among others. The production of superoxide by NOX activity in various allergenic pollens has been recently described. NOX activity has been shown to induce oxidative stress in cultured airway epithelium, driving to allergic inflammation. We report here the presence, molecular characteristics and expression of a NOX enzyme in olive pollen. Moreover, we describe the presence, activity and cellular localization of a well characterized allergenic protein from olive pollen (Ole e 5) displaying relevant homology with Cu/Zn SODs from different species, and a Mn-SOD also from olive pollen. The incidence of Ole e 5 in type I hypersensitivity reactions to olive pollen has been reported to be 35%, whereas several Mn-SODs have also been identified as allergens in different species. In addition to their intrinsic allergenic properties, both enzymes may regulate the levels of NOX-generated superoxide, therefore modulating the inflammation response to this pollen. A model of the interactions occurring among these molecules themselves, their substrates/products and the immune system is proposed., This work was supported by by ERDF-cofunded projects BFU2011-22779, P2010-AGR6274, P2010-CVI5767 and P2011-CVI-7487, Czech Science Foundation grant GACR 522/09/P299 to M.P. and a research agreement CSIC (Spain)-AVCR (Czech Republic) Ref. 2010CZ0001. MJ Jiménez-Quesada and A Zafra thanks the CSIC for JAE grant funding.

Trabajo presentado al 1er Congress of the Spanish Network of Advanced Optical Microscopy (REMOA), celebrado en Barcelona (España) del 19 al 21 de noviembre de 2012., Work supported by the ERDF co-funded projects BFU2008-00629 and BFU2011-22779.

Resumen del póster presentado al IV Spanish and Portuguese Meeting on Free Radicals, celebrado en Valencia del 5 al 7 de junio de 2012., The presence of high levels of NO has been described in several pollen grains, where their presence has been shown to regulate pollen tube growth and orientation. Here we have assessed the production of NO during in vitro germination in the olive pollen by using exogenous NO donors like sodium nitroprusside and S-nitrosoglutathione. Both chemical induced a remarkable inhibition of pollen germination and tube growth, which were partially reverted by the addition of the specific NO-scavenger carboxy-PTIO (cPTIO). The presence and localization of NO was determined either by using a fluorimetric assay and by confocal microscopy using the fluorescent probe diaminoflurescein-2 diacetate (DAF-2DA). As olive pollen is also a highly allergenic source, the putative role of NO in eliciting the hypersensitive response in humans is discussed., This work was supported by by ERDF-cofunded projects BFU2011-22779, P2010-AGR6274, P2010-CVI5767 and 2011-CVI-7487. MJ Jiménez-Quesada and A Zafra thanks the CSIC for JAE grant funding.

Resumen del trabajo presentado a las I Jornadas sobre Investigación Biomédica: Máster Universitario en Biotecnología y Biomedicina, celebradas el 20 de enero de 2012 en la Universidad de Jaén.

Trabajo presentado al 2nd Joint Congress of the Portuguese and Spanish Microscopy Societies: Microscopy at the Frontiers of Science, celebrado en España en octubre de 2011.

Resumen del trabajo presentado al 10th International Conference on Reactive Oxygen and Nitrogen Species in Plants celebrado en Budapest (Hungría) del 5 al 8 de Julio de 2011., This work was supported by research project BFU2008-00629 (Spanish MCINN) and a research agreement CSIC (Spain)-AVCR (Czech Republic) Ref. 2010CZ0001. A Zafra thanks the CSIC for JAE grant funding.

Trabajo presentado a las III Jornadas Nacionales del grupo de Horticultura de la SECH, celebradas en Sevilla el 6 y 7 de octubre de 2011.

Resumen del trabajo presentado al 10th International Conference on Reactive Oxygen and Nitrogen Species in Plants celebrado en Budapest (Hungría) del 5 al 8 de Julio de 2011.-- et al., This work was supported by research project BFU2008-00629 (Spanish MCINN), CVI-5767(Andalusian Government), Czech Science Foundation grant GACR 522/09/P299 to M.P. and a research agreement CSIC (Spain)-AVCR (Czech Republic) Ref. 2010CZ0001. MJ Jiménez-Quesada and A Zafra thanks the CSIC for JAE grant funding.

pp:607-613 Publisher: Formatex Research Center, Several components of the exine (i.e. the outer layer of the pollen wall) show conspicuous autofluorescence in the pollen grain. We describe the autofluorescence properties of pollens from the Mediterranean cypress, lily, turnip and olive (cvs. Picual, Loaime, Cornicabra and Ayrouni). The intensity and pattern of autofluorescence largely varied depending on the species and the hydration stage of pollen. In the case of olive, differences in the level of autofluorescence were detected among the four cultivars analyzed. This technique also allowed further morphological characterization of pollen (e.g. determination of pollen size and shape, and the number and type of apertures, the pattern of the exine, etc), evaluation of pollen integrity, detection of additional components (e.g. the pollen coat) over the surface of the pollen wall, etc. The methods used can be considered non-disruptive and lack of sample preparation. Monitoring pollen autofluorescence could be useful in disciplines like Aerobiology, Plant Taxonomy, and Cell and Molecular Biology.

Resumen de la comunicación presentada al 28º International Horticultural Congress celebrado del 22 al 27 de 2010 en Lisboa.-- Sm09.012.

Trabajo presentado al 3rd Plant NO Club International Meeting, celebrado en la República Checa del 15 al 16 de julio de 2010.

Resumen de la comunicación oral presentada en el XVII Congress of the Federation of European Societies of Plant Biology (FESPB), celebrado del 4 al 9 de Julio de 2010 en Valencia.-- P04-010., The presence of reactive oxygen species (ROS) and NO in the reproductive tissues of several Angiosperms has been investigated. Early studies have shown that these molecules play putative roles as signaling molecules during the interaction pollen-pistil. However, their occurrence in other reproductive phases has not been investigated. We have analyzed the presence of ROS and NO in floral tissues over different developmental stages (unopened flower buds, recently opened flowers, dehiscent anthers and flowers after fertilization) by using different fluorophores and confocal laser scanning microscopy. The study was carried out in species with different types of stigmas and systems of compatibility (olive tree, orange, tomato, pepper, pea and A. thaliana). Although the presence of ROS in the stigmas was detected at higher levels during those developmental phases considered “receptive” to pollen interaction, these molecules were also present at early (unopened flower) or later (post-fertilization) stages. The biological significance of the presence of these products may differ between these stages, including defence functions, signaling and senescence. The study confirms the enhanced production of NO by pollen grains and tubes during the receptive phase, and the decrease in the presence of ROS when NO is actively produced., This work was supported by MICINN (project BFU2008-00629/BFI), and by JA (project P06-AGR-01791).

2 páginas.-- Resumen del poster presentado en el XXI International Congress on Sexual Plant Reproduction, celebrado del 2 al 6 de Agosto de 2010 en Bristol.-- PSR4.19., Work supported by FEDER co-financed projects BFU2008-00629/BFI and P06-AGR-01791.

1 página.-- Resumen de la comunicación presentada en el XXI International Congress on Sexual Plant Reproduction, celebrado del 2 al 6 de Agosto de 2010 en Bristol.-- PSR4.7., This work was supported by FEDER co-funded research projects P06-AGR-01719 (Junta de Andalucía) and BFU2008-00629 (MCINN).

Resumen del trabajo presentado al "Plant Oxygen Group Meeting on Reactive Oxygen and Nitrogen Species" celebrado en Helsinki (Finlandia) del 8 al 10 de Julio de 2009., This work was funded by Projects P06-AGR-01791 and BFU2008-00629 (CICE-JA and PN I+D+I, respectively).

Resumen del trabajo presentado al "Biochemical Society Focused Meeting" celebrado en Bath (Reino Unido) del 14 al 16 de Septiembre de 2009., Work supported by projects P06-AGR-01719 and BFU2008-00629.

Resumen del trabajo presentado a la XVIII Reunión de la Sociedad Española de Fisiología Vegetal (SEFV) y al XI Congreso Hispano-Luso de Fisiología Vegetal, S2-P24: Crecimiento y desarrollo, celebrados en Zaragoza (España) del 8 al 11 de Septiembre de 2009., This work was supported by research projects BFU2008-00629 (Spanish MCyT) and P06-AGR-01719 (Andalusian Regional Government). A Zafra thanks the CSIC for providing JAE grant funding.

Resumen del trabajo presentado a la XVIII Reunión de la Sociedad Española de Fisiología Vegetal (SEFV) y al XI Congreso Hispano-Luso de Fisiología Vegetal, S2-P9: Crecimiento y desarrollo, celebrados en Zaragoza (España) del 8 al 11 de Septiembre de 2009., This work was supported by research projects BFU2008-00629 (MCeI) and P06-AGR-01719 (Junta de Andalucía). A. Zafra thanks the CSIC for providing a JAE grant.

3 páginas.-- Comunicacion presentada al XX International Congress on Sexual Plant Reproduction, celebrado del 4 al 8 de 2008 en Brasilia (Brasil)., This work was funded by MEC project BFU2004-00601/BFI and by "Consejería de Innovación, Ciencia y Empresa de la Junta de Andalucía", project PO6-AGr-01791.

5 páginas, 3 figuras.-- Comunicación presentada a las VI Jornadas Internacionales de Olivar Ecológico (Ecoliva 2007)., El olivo es una planta de gran importancia agronómica en los países mediterráneos, y muy especialmente en nuestra región. Nuestro grupo viene tradicionalmente estudiando numerosos aspectos de la reproducción sexual de dicha especie, tales como el desarrollo del polen, sus características (viabilidad y capacidad de germinación), el desarrollo del gineceo, los procesos de fertilización, la formación y maduración del fruto, etc. Para estos estudios utilizamos distintos métodos que incluyen desde la observación morfológica mediante técnicas de microscopía óptica, microscopía electrónica de transmisión, barrido y microscopía confocal, hasta técnicas de aislamiento y caracterización de proteínas y genes implicados en dichos procesos. Todos estos estudios nos están ayudando a comprender el sistema de reproducción sexual del olivo, una planta en la que la presencia de mecanismos de autoincompatibilidad polen-pistilo con bases aparentemente genéticas y ambientales, está empezando a ser estudiada. Otra línea de investigación en curso en nuestro grupo es aquella que se refiere al estudio de determinadas proteínas del grano de polen con funciones clave en su desarrollo y en la germinación y crecimiento del tubo polínico, y que además exhiben carácter alergénico para algunos individuos. Las características moleculares y el contenido de estos alérgenos varían significativamente entre cultivares de olivo, lo cual hace especialmente interesante su estudio en la fisiología del grano de polen, además de las implicaciones clínicas que de ello se derivan., Los resultados descritos han sido obtenidos gracias a la financiación de diversas instituciones públicas, esencialmente, los proyectos del Plan Nacional de I+D+i mencionados anteriormente y varios planes de formación de personal investigador de dicho programa, así como de la AECI y del programa I3P-CSIC.

7 páginas, 4 figuras., [EN] The olive tree is grown in many parts of the world. Its germplasm is very broad, with 250 varieties in Spain alone. Variations in the ability of pollen to germinate have been studied in detail and show conspicuous differences between varieties. However, commercial olive pollen from cultivars whose origin is unknown is the material that is commonly used for clinical and biological studies. We aim to assess the putative heterogeneity of olive cultivars with regard to the presence of several pollen allergens and to determine whether these differences have biological and clinical relevance. Previous studies show that most allergens isolated and characterized to date are highly polymorphic. Olive cultivars display wide differences in the expression levels of many allergens and in the number and molecular characteristics of the allergen isoforms expressed. These differences are maintained over the years, and are intrinsic to the genetics of each cultivar. Such broad polymorphism seems to be involved in the physiology of the olive reproductive system, which might include the adaptation of the plant to different environmental conditions, the establishment of the compatibility system, and pollen performance. The differences in allergen composition in cultivars, particularly in the Ole e 1 allergen, are responsible for the important differences in the allergenic potency of the extracts. These fi ndings could have a number of implications for the diagnosis and therapy of olive pollen allergy. We discuss how cultivar differences affect extract quality, diagnostic and therapeutic effi cacy and safety, and the development of new vaccines based on the use of recombinant allergens., [ES] El olivo es un cultivo ampliamente representado en el mundo. Su germoplasma es muy amplio, con 250 variedades sólo en España. La capacidad del polen para germinar, que presenta notables diferencias entre variedades, ha sido estudiada en detalle. El material usado comúnmente para estudios clínicos y biológicos es, sin embargo, polen comercial de cultivares de origen desconocido. Nuestro objetivo es evaluar la posible heterogeneidad de los cultivares de olivo en relación a la presencia de varios alérgenos del polen, y determinar si esas diferencias tienen relevancia biológica y clínica. Estudios previos muestran que la mayor parte de los alérgenos aislados y caracterizados hasta la fecha son altamente polimórfi cos. Los cultivares de olivo muestran amplias diferencias en los niveles de expresión de muchos alérgenos, así como en el número y características moleculares de las isoformas alergénicas expresadas. Estas diferencias se mantienen a lo largo de años, y son intrínsecas a la genética de cada cultivar. Este amplio polimorfi smo parece estar implicado en la fi siología del sistema reproductivo del olivo, en relación con la adaptación de la planta a diferentes condiciones ambientales, el establecimiento de un sistema de compatibilidad, y el dinamismo del polen. Las diferencias en la composición alergénica de los cultivares, particularmente en cuanto al alérgeno Ole e 1, son responsables de las importantes diferencias en la potencia alergénica de los extractos. Estos hallazgos pueden tener numerosas implicaciones en la diagnosis y terapia de la alergia al polen del olivo. Discutimos cómo las diferencias entre cultivares afectan a la calidad del polen, a la efi cacia y seguridad del diagnóstico y la terapia, así como al desarrollo de nuevas vacunas basadas en el uso de alérgenos recombinantes.

Trabajo presentado a las I Jornadas del Grupo de Olivicultura de la Sociedad Española de Ciencias Hortícolas (SECH) celebradas en Córdoba (España) del 9 al 10 de Octubre de 2006., Este trabajo ha sido financiado por los proyectos AGL2003-00719 y MEC BFU2004-00601/BFI.