Gene-Metabolite Networks of Volatile Metabolism in Airen and Tempranillo Grape Cultivars Revealed a Distinct Mechanism of Aroma Bouquet Production

Volatile compounds are the major determinants of aroma and flavor in both grapes and wine. In this study, we investigated the emission of volatile and non-volatile compounds during berry maturation in two grape varieties (Airén and Tempranillo) throughout 2010 and 2011. HS-SPME coupled to gas chromatography and mass spectrometry was applied for the identification and relative quantitation of these compounds. Principal component analysis was performed to search for variability between the two cultivars and evolution during 10 developmental stages. Results showed that there are distinct differences in volatile compounds between cultivars throughout fruit development. Early stages were characterized in both cultivars by higher levels of some apocarotenoids such as β-cyclocitral or β-ionone, terpenoids (E)-linalool oxide and (Z)-linalool oxide and several furans, while the final stages were characterized by the highest amounts of ethanol, benzenoid phenylacetaldehyde and 2-phenylethanol, branched-amino acid-derived 3-methylbutanol and 2-methylbutanol, and a large number of lipid derivatives. Additionally, we measured the levels of the different classes of volatile precursors by using liquid chromatography coupled to high resolution mass spectrometry. In both varieties, higher levels of carotenoid compounds were detected in the earlier stages, zeaxanthin and α-carotene were only detected in Airén while neoxanthin was found only in Tempranillo; more variable trends were observed in the case of the other volatile precursors. Furthermore, we monitored the expression of homolog genes of a set of transcripts potentially involved in the biosynthesis of these metabolites, such as some glycosyl hydrolases family 1, lipoxygenases, alcohol dehydrogenases hydroperoxide lyases, O-methyltransferases and carotenoid cleavage dioxygenases during the defined developmental stages. Finally, based on Pearson correlation analyses, we explored the metabolite-metabolite fluctuations within VOCs/precursors during the berry development; as well as tentatively linking the formation of some metabolites detected to the expression of some of these genes. Our data showed that the two varieties displayed a very different pattern of relationships regarding the precursor/volatile metabolite-metabolite fluctuations, being the lipid and the carotenoid metabolism the most distinctive between the two varieties. Correlation analysis showed a higher degree of overall correlation in precursor/volatile metabolite-metabolite levels in Airén, confirming the enriched aroma bouquet characteristic of the white varieties.This work was supported by the “Junta de comunidades de Castilla-La Mancha” (JCCM) [PPII10-0062-7718] and benefited from the networking activities within the European Cooperation in Science and Technology Action CA15136 (EUROCAROTEN). GD was supported by short-term fellowships of the Quality Fruit (FA1106) European Cooperation in Science and Technology actions. OA was funded by FPCYTCLM through the INCRECYT Programme.Peer reviewedPeer Reviewe

Fortification and bioaccessibility of saffron apocarotenoids in potato tubers

Carotenoids are C40 isoprenoids with well-established roles in photosynthesis, pollination, photoprotection, and hormone biosynthesis. The enzymatic or ROS-induced cleavage of carotenoids generates a group of compounds named apocarotenoids, with an increasing interest by virtue of their metabolic, physiological, and ecological activities. Both classes are used industrially in a variety of fields as colorants, supplements, and bio-actives. Crocins and picrocrocin, two saffron apocarotenoids, are examples of high-value pigments utilized in the food, feed, and pharmaceutical industries. In this study, a unique construct was achieved, namely O6, which contains CsCCD2L, UGT74AD1, and UGT709G1 genes responsible for the biosynthesis of saffron apocarotenoids driven by a patatin promoter for the generation of potato tubers producing crocins and picrocrocin. Different tuber potatoes accumulated crocins and picrocrocin ranging from 19.41-360 to 105-800 mu g/g DW, respectively, with crocetin, crocin 1 [(crocetin-(beta-D-glucosyl)-ester)] and crocin 2 [(crocetin)-(beta-D-glucosyl)-(beta-D-glucosyl)-ester)] being the main compounds detected. The pattern of carotenoids and apocarotenoids were distinct between wild type and transgenic tubers and were related to changes in the expression of the pathway genes, especially from PSY2, CCD1, and CCD4. In addition, the engineered tubers showed higher antioxidant capacity, up to almost 4-fold more than the wild type, which is a promising sign for the potential health advantages of these lines. In order to better investigate these aspects, different cooking methods were applied, and each process displayed a significant impact on the retention of apocarotenoids. More in detail, the in vitro bioaccessibility of these metabolites was found to be higher in boiled potatoes (97.23%) compared to raw, baked, and fried ones (80.97, 78.96, and 76.18%, respectively). Overall, this work shows that potatoes can be engineered to accumulate saffron apocarotenoids that, when consumed, can potentially offer better health benefits. Moreover, the high bioaccessibility of these compounds revealed that potato is an excellent way to deliver crocins and picrocrocin, while also helping to improve its nutritional value

The francoists television newscasts. An investigation on the sources

Introduction. A major drawback of the existing studies on the history of television in Spain under Franco is their failure to analyse direct sources. Objectives. The aim of this research is to analyse TVE´s telediarios (television news bulletins) during the Franco era, using new and specific data. Method. Rather than using them purely as examples, this research makes comprehensive use of the programme schedules found in La Vanguardia, Abc and Tele Radio, as well as TVE’s official broadcast reports. Conclusions. Through the use of these sources, this paper intends to demonstrate the availability of a wide range of resources, on the basis of which there is a need for new critical studies on the history of television in Spain to be carried out.Introducción. Los estudios sobre historia de la televisión en España durante el franquismo no recurren a las fuentes directas. Objetivos. El objetivo de esta investigación es analizar desde datos nuevos y concretos, los telediarios de TVE en el periodo franquista. Método. Para el desarrollo de esta investigación se han empleado de manera exhaustiva, no como mero ejemplo, las parrillas de programación recuperadas de La Vanguardia, Abc y Tele Radio, y los partes de emisión de TVE. Conclusiones. Con el manejo de estas fuentes se pretende mostrar la existencia de una amplia gama de recursos de información accesibles, desde las que es preciso abordar nuevos estudios críticos de historia de la televisión en España

Plantes transgéniques produisant des niveaux élevés de composés d'apocaroténoïdes et leurs utilisations

The invention provides methods and materials for recombinantly producing apocarotenoids from transgenic plants, and more particularly, tomato plants, expressing heterologous genes from Crocus sativus, the saffron plant, under the control of specific promoters. Moreover, the present disclosure also relates to the genetic constructs comprising the heterologous genes, and the use thereof for the expression in plants, preferably tomatoes, and to a method for obtaining transgenic pants, preferably tomatoes plants, which express the genetic construct mentioned above, and which have a high concentration of apocarotenoids products, in particular crocins and picrocrocins. Furthermore, the present disclosure also refers to the food products obtained from transgenic plants, preferably tomatoes and/or tomato-based products, nutraceutical and pharmaceutical compositions comprising thereof and the use of such compositions as medicine, preferably for the treatment of neurodegenerative diseases such as Alzheimer's disease.Peer reviewedUniversidad de Castilla-La Mancha, Consejo Superior de Investigaciones Científicas, Universitat Politècnica de ValènciaA1 Solicitud de patente con informe sobre el estado de la técnic

Plantes transgéniques produisant des niveaux élevés de composés d'apocaroténoïdes et leurs utilisations

[EN] The invention provides methods and materials for recombinantly producing apocarotenoids from transgenic plants, and more particularly, tomato and potato plants, expressing heterologous genes from Crocus sativus, the saffron plant, under the control of specific promoters. Moreover, the present disclosure also relates to the genetic constructs comprising the heterologous genes, and the use thereof for the expression in plants and to a method for obtaining transgenic plants which express the genetic construct mentioned above, and which have a high concentration of apocarotenoids products, in particular crocins and picrocrocins. Furthermore, the present disclosure also refers to the food products obtained from such transgenic plants, nutraceutical and pharmaceutical compositions comprising thereof and the use of such compositions as medicine, preferably for the treatment of neurodegenerative diseases such as Alzheimer's disease.[FR] L'invention concerne des procédés et des matériaux pour produire par recombinaison des apocaroténoïdes à partir de plantes transgéniques, et plus particulièrement, des plants de tomate et de pomme de terre, exprimant des gènes hétérologues à partir de Crocus sativus, le plant de safran, sous le contrôle de promoteurs spécifiques. De plus, la présente invention concerne également les constructions génétiques comprenant les gènes hétérologues, et leur utilisation pour l'expression dans des plantes et un procédé d'obtention de plantes transgéniques qui expriment la construction génétique mentionnée ci-dessus, et qui ont une concentration élevée en produits apocaroténoïdes, en particulier des crocines et des picrocrocines. En outre, la présente invention concerne également les produits alimentaires obtenus à partir de telles plantes transgéniques, des compositions nutraceutiques et pharmaceutiques les comprenant et l'utilisation de telles compositions en tant que médicament, de préférence pour le traitement de maladies neurodégénératives telles que la maladie d'Alzheimer.Peer reviewedUniversidad de Castilla-La Mancha, Consejo Superior de Investigaciones Científicas, Universitat Politècnica de ValènciaA1 Solicitud de patente con informe sobre el estado de la técnic

Expression and Interaction Analysis among Saffron ALDHs and Crocetin Dialdehyde

In saffron, the cleavage of zeaxanthin by means of CCD2 generates crocetin dialdehyde, which is then converted by an unknown aldehyde dehydrogenase to crocetin. A proteome from saffron stigma was released recently and, based on the expression pattern and correlation analyses, five aldehyde dehydrogenases (ALDHs) were suggested as possible candidates to generate crocetin from crocetin dialdehydes. We selected four of the suggested ALDHs and analyzed their expression in different tissues, determined their activity over crocetin dialdehyde, and performed structure modeling and docking calculation to find their specificity. All the ALDHs were able to convert crocetin dialdehyde to crocetin, but two of them were stigma tissue-specific. Structure modeling and docking analyses revealed that, in all cases, there was a high coverage of residues in the models. All of them showed a very close conformation, indicated by the low root-mean-square deviation (RMSD) values of backbone atoms, which indicate a high similarity among them. However, low affinity between the enzymes and the crocetin dialdehyde were observed. Phylogenetic analysis and binding affinities calculations, including some ALDHs from Gardenia jasmonoides, Crocus sieberi, and Buddleja species that accumulate crocetin and Bixa orellana synthetizing the apocarotenoid bixin selected on their expression pattern matching with the accumulation of either crocins or bixin, pointed out that family 2 C4 members might be involved in the conversion of crocetin dialdehyde to crocetin with high specificity

Data_Sheet_1_Fortification and bioaccessibility of saffron apocarotenoids in potato tubers.docx

Carotenoids are C40 isoprenoids with well-established roles in photosynthesis, pollination, photoprotection, and hormone biosynthesis. The enzymatic or ROS-induced cleavage of carotenoids generates a group of compounds named apocarotenoids, with an increasing interest by virtue of their metabolic, physiological, and ecological activities. Both classes are used industrially in a variety of fields as colorants, supplements, and bio-actives. Crocins and picrocrocin, two saffron apocarotenoids, are examples of high-value pigments utilized in the food, feed, and pharmaceutical industries. In this study, a unique construct was achieved, namely O6, which contains CsCCD2L, UGT74AD1, and UGT709G1 genes responsible for the biosynthesis of saffron apocarotenoids driven by a patatin promoter for the generation of potato tubers producing crocins and picrocrocin. Different tuber potatoes accumulated crocins and picrocrocin ranging from 19.41–360 to 105–800 μg/g DW, respectively, with crocetin, crocin 1 [(crocetin-(β-D-glucosyl)-ester)] and crocin 2 [(crocetin)-(β-D-glucosyl)-(β-D-glucosyl)-ester)] being the main compounds detected. The pattern of carotenoids and apocarotenoids were distinct between wild type and transgenic tubers and were related to changes in the expression of the pathway genes, especially from PSY2, CCD1, and CCD4. In addition, the engineered tubers showed higher antioxidant capacity, up to almost 4-fold more than the wild type, which is a promising sign for the potential health advantages of these lines. In order to better investigate these aspects, different cooking methods were applied, and each process displayed a significant impact on the retention of apocarotenoids. More in detail, the in vitro bioaccessibility of these metabolites was found to be higher in boiled potatoes (97.23%) compared to raw, baked, and fried ones (80.97, 78.96, and 76.18%, respectively). Overall, this work shows that potatoes can be engineered to accumulate saffron apocarotenoids that, when consumed, can potentially offer better health benefits. Moreover, the high bioaccessibility of these compounds revealed that potato is an excellent way to deliver crocins and picrocrocin, while also helping to improve its nutritional value.</p