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[Background] Climate and land-use changes, which include the application of various types of organic and inorganic fertilisers, have been reducing the species diversity of Mediterranean grasslands and threatening its conservation. Annual plants are one of the most diverse functional groups of species in these grasslands, despite suffering competitive pressure from perennial herbaceous and woody species, and they are essential for ecosystem functioning and stability., [Aims] To quantify how fertilisation modulates the impact of plant-to-plant interactions and climate fluctuations on the dynamics of annuals in Mediterranean grasslands. We hypothesised that the application of sewage sludge would increase competition between functional groups, reducing the abundance of annuals in the long-term, but would buffer the negative impacts of drought on the year-to-year fluctuation of the diversity of annuals., [Methods] In a semi-natural species-rich Mediterranean grassland in northern Spain, we analysed the changes in the taxonomical and functional composition and diversity of annuals over 14 years in response to variations in the abundance of perennial herbaceous and woody species, climate fluctuations and fertilisation with sewage sludge. We quantified separately the patterns of year-to-year fluctuations and long-term trends., [Results] The frequency and diversity of annuals decreased with higher abundance of perennial herbaceous species, drought in June and cold winters. The addition of sewage sludge decreased the abundance of annuals in the long-term, seemed to promote competition between annuals and other functional groups at an interannual scale, and mitigated the negative effects of drought and cold., [Conclusions] Fertilisation influences differently the temporal response of annuals to climate fluctuations and plant-to-plant interactions., This work was supported by Fundación Caja Navarra under Grant Programa ‘Tú Eliges, Tú Decides’, ref. 10833; and by Universidad de Navarra under Projects ‘Biodiversity Data Analytics and Environmental Quality’ and ‘Red de Observatorios de la Biodiversidad de Navarra (ROBIN)’. M.V. was supported by the Asociación de Amigos de la Universidad de Navarra under Grant Convocatoria de Ayudas para la Formación del Personal Investigador de la Asociación de Amigos de la Universidad de Navarra para el curso 2018-2019 (September 2018-March 2019); and the Departamento de Educación del Gobierno de Navarra under Grant Ayudas predoctorales para la realización de programas de doctorado de interés para Navarra, Plan de Formación y de I+D 2018. A.G. was supported by Ministerio de Ciencia e Innovación under Grant RyC2020-030647-I; and the CSIC under Grant PIE-20223AT003.

14 pags.- 4 Figs.- 1 Tabl. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license., empranillo Blanco is a somatic variant of Tempranillo Tinto that appeared as a natural, spontaneous mutation in 1988 in a single shoot of a single plant in an old vineyard. It was vegetatively propagated, and currently wines from Tempranillo Blanco are commercially available. The mutation that originated Tempranillo Blanco comprised single-nucleotide variations, chromosomal deletions, and reorganizations, losing hundreds of genes and putatively affecting the functioning and regulation of many others. The most evident, visual change in Tempranillo Blanco is the anthocyanin lost, producing this grapevine variety bunches of colorless grapes. This review aims to summarize from the available literature differences found between Tempranillo Blanco and Tinto in addition to the color of the grapes, in a climate change context and using fruit-bearing cuttings grown in temperature-gradient greenhouses as research-oriented greenhouses. The differences found include changes in growth, water use, bunch mass, grape quality (both technological and phenolic maturity), and some aspects of their photosynthetic response when grown in an atmosphere of elevated CO2 concentration and temperature, and low water availability. Under field conditions, Tempranillo Blanco yields less than Tempranillo Tinto, the lower weight of their bunches being related to a lower pollen viability and berry and seed setting., Authors acknowledge Innovine European project (Nº 311775), Aragón Government (A03 group) and “Ministerio de Ciencia e Innovación” of Spain (MCINN AGL2014-56075-C2-1-R) for funding and “Asociación de Amigos de la Universidad de Navarra” for T. Kizildeniz grant.

Due to the CO greenhouse effect, elevated atmospheric concentration leads to higher temperatures, accompanied by episodes of less water availability in semiarid and arid areas or drought periods. Studies investigating these three factors (CO, temperature and water availability) simultaneously in grapevine are scarce. The present work aims to analyze the combined effects of high CO (700 ppm), high temperature (ambient +4°C) and drought on the photosynthetic activity, biomass allocation, leaf non-structural carbon composition, and carbon/nitrogen (C/N) ratio in grapevine. Two grapevine cultivars, red berry Tempranillo and white berry Tempranillo, were used, the latter being a natural, spontaneous mutant of the red cultivar. The experiment was performed on fruit-bearing cuttings during a 3-month period, from June (fruit set) to August (maturity). The plants were grown in research-oriented facilities, temperature-gradient greenhouses, where temperature, CO, and water supply can be modified in a combined way. Drought had the strongest effect on biomass accumulation compared to the other environmental variables, and root biomass allocation was increased under water deficit. CO and temperature effects were smaller and depended on cultivar, and on interactions with the other factors. Acclimation effects were observed on both cultivars as photosynthetic rates under high atmospheric CO were reduced by long-term exposition to elevated CO. Exposure to such high CO resulted in increased starch concentration and reduced C/N ratio in leaves. A correlation between the intensity of the reduction in photosynthetic rates and the accumulation of starch in the leaves was found after prolonged exposure to elevated CO., “Asociación de Amigos de la Universidad de Navarra”, Grant/Award Number: T Kizildeniz grant; “Ministerio de Ciencia e Innovación” of Spain, Grant/Award Number: MCINN AGL2014-56075-C2-1-R; Aragón Government, Grant/Award Number: A03 group; Innovine European project, Grant/Award Number: 311775

Background and Aims: The interactive effects of simulated 2100 environmental conditions (air temperature and CO level) and water deficit (WD) on four clones of Vitis vinifera cv. Tempranillo were investigated. Methods and Results: Fruit-bearing cuttings were subjected to: (i) two temperature/CO/relative humidity (RH) regimes: climate change (28°C/18°C, 700 μmol/mol CO and 33%/53% RH, day/night) versus current climatic conditions (24°C/14°C, 400 μmol/mol CO and 45%/65% RH), combined with (ii) two water availabilities: well-watered (WW) versus WD. Climate change increased net photosynthesis (A), transiently ameliorating the low carbon fixation rates under drought, but not the reduction in vegetative and reproductive growth. Climate change increased intrinsic water use efficiency (A/g), especially when combined with WD, but not the instantaneous water use efficiency (A/T). The clones exhibited differences in the ripening time, plant vigour and reproductive growth. Variability in the response of A, phenology and growth to the simulated conditions was observed among clones. Conclusions: Differences in the length of the reproductive cycle conditioned, in part, the physiological response of the clones to the environmental factors. Significance of the Study: The study improves our understanding of the interactive effects of climate change factors and provides insights into the response of different clones, as the basis for the adaptation of cultivars in their traditional growing regions. This work was supported by the Ministerio de Economía y Competitividad of Spain [AGL2014-56075-C2-1-R]; Fundación Universitaria de Navarra [PIUNA 2018] and Asociación de Amigos de la Universidad de Navarra (grant to Dr Marta Arrizabalaga-Arriazu).

Background and Aims: The interactive effects of simulated 2100 environmental conditions (air temperature and CO level) and water deficit (WD) on four clones of Vitis vinifera cv. Tempranillo were investigated. Methods and Results: Fruit-bearing cuttings were subjected to: (i) two temperature/CO/relative humidity (RH) regimes: climate change (28°C/18°C, 700 μmol/mol CO and 33%/53% RH, day/night) versus current climatic conditions (24°C/14°C, 400 μmol/mol CO and 45%/65% RH), combined with (ii) two water availabilities: well-watered (WW) versus WD. Climate change increased net photosynthesis (A), transiently ameliorating the low carbon fixation rates under drought, but not the reduction in vegetative and reproductive growth. Climate change increased intrinsic water use efficiency (A/g), especially when combined with WD, but not the instantaneous water use efficiency (A/T). The clones exhibited differences in the ripening time, plant vigour and reproductive growth. Variability in the response of A, phenology and growth to the simulated conditions was observed among clones. Conclusions: Differences in the length of the reproductive cycle conditioned, in part, the physiological response of the clones to the environmental factors. Significance of the Study: The study improves our understanding of the interactive effects of climate change factors and provides insights into the response of different clones, as the basis for the adaptation of cultivars in their traditional growing regions., This work was supported by the Ministerio de Economía y Competitividad of Spain [AGL2014-56075-C2-1-R]; Fundación Universitaria de Navarra [PIUNA 2018] and Asociación de Amigos de la Universidad de Navarra (grant to Dr Marta Arrizabalaga-Arriazu).

18 Pags.- 6 Figs.- 2 Tabls. Copyright © 2020 Arrizabalaga-Arriazu, Gomès, Morales, Irigoyen, Pascual and Hilbert. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., Tempranillo is a grapevine (Vitis vinifera L.) variety extensively used for world wine production which is expected to be affected by environmental parameters modified by ongoing global climate changes, i.e., increases in average air temperature and rise of atmospheric CO2 levels. Apart from determining their effects on grape development and biochemical characteristics, this paper considers the intravarietal diversity of the cultivar Tempranillo as a tool to develop future adaptive strategies to face the impact of climate change on grapevine. Fruit-bearing cuttings of five clones (RJ43, CL306, T3, VN31, and 1084) were grown in temperature gradient greenhouses (TGGs), from fruit set to maturity, under two temperature regimes (ambient temperature vs. ambient temperature plus 4°C) and two CO2 levels (ambient, ca. 400 ppm, vs. elevated, 700 ppm). Treatments were applied separately or in combination. The analyses carried out included berry phenological development, the evolution in the concentration of must compounds (organic acids, sugars, and amino acids), and total skin anthocyanins. Elevated temperature hastened berry ripening, sugar accumulation, and malic acid breakdown, especially when combined with high CO2. Climate change conditions reduced the amino acid content 2 weeks after mid-veraison and seemed to delay amino acidic maturity. Elevated CO2 reduced the decoupling effect of temperature on the anthocyanin to sugar ratio. The impact of these factors, taken individually or combined, was dependent on the clone analyzed, thus indicating certain intravarietal variability in the response of Tempranillo to these climate change-related factors., This work was supported by the Ministerio de Economía y Competitividad of Spain (AGL2014-56075-C2-1-R), Fundación Universitaria de Navarra (2018), European Union (Erasmus+ grant to MA-A), Aquitaine Regional Council (AquiMOb grant to MA-A), and Asociación de Amigos de la Universidad de Navarra (doctoral grant to MA-A). This work also supported by the metaprogramme Adaptation of Agriculture and Forests to Climate Change (AAFCC) of the French National Institute for Agriculture, Food and Environment (INRAE), in the frame of LACCAVE2-21 project.

Atmospheric CO2 levels and global temperatures are expected to rise in the next decades, and viticulture must face these changes. Within this context, exploiting the intra-varietal diversity of grapevine (Vitis vinifera L.) can be a useful tool for the adaptation of this crop to climate change. The aim of the present work was to study the effect of elevated temperature and elevated levels of atmospheric CO2, both individually and combined, on the growth, phenology and carbon partitioning of five clones of the cultivar Tempranillo (RJ43, CL306, T3, VN31 and 1084). The hypothesis that clones within the same variety that differ in their phenological development may respond in a different manner to the above mentioned environmental factors from a physiological point of view was tested. Grapevine fruit-bearing cuttings were grown from fruit set to maturity under two temperature regimes: ambient (T) vs elevated (ambient + 4°C, T + 4), combined with two CO2 levels: ambient (ca. 400 ppm, ACO2) vs elevated (700 ppm, ECO2), in temperature-gradient greenhouses (TGGs). Considering all the clones, elevated temperature hastened grape development and increased vegetative growth, but reduced grape production, the later most likely associated with the heat waves recorded during the experiment. Plants in the elevated CO2 treatments showed a higher photosynthetic activity at veraison and an increased vegetative growth, but they showed signs of photosynthetic acclimation to ECO2 at maturity according to the C:N ratio, especially when combined with high temperature. The combination of ECO2 and T + 4, mimicking climate change environmental conditions, showed additive effects in some of the parameters analyzed. The clones showed differences in their phenological development, which conditioned some responses to elevated CO2 and temperature in terms of vegetative production and C partitioning into different organs. The work adds new knowledge on the use of different grapevine clones, that can be useful to improve the viticultural efficiency in future climate change scenarios., This work was funded by the Ministerio de Economía y Competitividad of Spain (AGL2014-56075-C2-1-R) and Asociación de Amigos de la Universidad de Navarra (grant to M. Arrizabalaga).

11 Pags.- 4 Tabls.- 6 Figs., In recent decades, agricultural production is being affected by a sharp increase in atmospheric CO2 concentration. Due to the greenhouse effect gases, crops are impacted by enhanced temperatures and concomitantly by increased scarce water availability. All arid and semiarid areas, including Mediterranean viticulture, must face these three climate change-related factors: atmospheric CO2 concentration and temperature increases, and scarce water for irrigation. Scarce water is a problem even in irrigated viticulture, as irrigation is becoming more and more restricted. Within this context, the aim of this work was to investigate grapevine (Vitis vinifera L. cv. red and white Tempranillo) vegetative and reproductive growth. Fruit-bearing cuttings were grown under elevated CO2 (around 700 μmol mol−1 or ppm, versus 400), high temperature (ambient temperature + 4 °C, versus ambient) and water deficit (cyclic drought, versus full irrigated) in temperature gradient greenhouses for three consecutive growing seasons (years 2013, 2014 and 2015). Climate change impacted markedly vegetative growth. Within the abovementioned factors, vegetative growth (total vegetative mass) was significantly reduced by drought (consistent the three years) and was associated to a low substrate water status and low leaf stomatal conductance. Elevated CO2 stimulated total vegetative mass, whereas leaf area was not affected. When plants were grown under elevated CO2, the largest increases were observed in leaf (white) and root (red) growth. In these experiments, elevated CO2 did not compensate the negative effects of water stress. An increase of the mean temperature 4 °C had no consequences on vegetative growth. Yield and yield-related traits were unaffected by the climate change scenario., Authors acknowledge Innovine European project (N° 311775), Aragón Government (A03 group) and “Ministerio de Ciencia e Innovación” of Spain (MCINN AGL2014-56075-C2-1-R) for funding and “Asociación de Amigos de la Universidad de Navarra” for T. Kizildeniz grant.

Telomere shortening with age has been documented in many organisms, butfew studies have reported telomere length measurements in amphibians, andno information is available for growth after metamorphosis, nor in wild popu-lations. We provide both cross-sectional and longitudinal evidence of nettelomere attrition with age in a wild amphibian population of natterjacktoads (Epidalea calamita). Based on age-estimation by skeletochronology andqPCR telomere length measurements in the framework of an individual-based monitoring programme, we confirmed telomere attrition in recapturedmales. Our results support that toads experience telomere attrition throughouttheir ontogeny, and that most attrition occurs during the first 1–2 years. We didnot find associations between telomere length and inbreeding or body con-dition. Our results on telomere length dynamics under natural conditionsconfirm telomere shortening with age in amphibians and provide quantifi-cation of wide telomere length variation within and among age-classes in awild breeding population., This work was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades, Spain (grant nos CGL2017-83131-P, CGL2017-83407-P) and Asociación de Amigos de la Universidad de Navarra (predoctoral grant to G.S.-M.).

Resumen del trabajo presentado en el 1st International Congress on Biosystems Engineering (ICOBEN), celebrado en Hatay (Turquía), del 24 al 27 de septiembre de 2019, Atmospheric CO2 concentration has increased from preindustrial period values of 280 ¿mol CO2 mol-1 air (ppm) to nowadays more than 400 ppm, and it is expected to reach 700 ppm by the end of this century. Due to its greenhouse effect, high CO2 concentration leads to higher temperatures that are being accompanied by less water availability. The aim of this study was to evaluate the changes in red and white Tempranillo grapevine vegetative and reproductive growth affected by the three climate change-related factors above mentioned, the water deficit, elevated CO2 and elevated temperature. Grapevine fruit-bearing cuttings were grown under elevated CO2, elevated temperature and drought conditions in temperature gradient greenhouses located at the University of Navarra (Pamplona, Spain) during three consecutive growing seasons (years 2013, 2014 and 2015) with the aim of detecting individual and/or combined effects of climate change-related factors in the Tempranillo white and red varieties. Drought reduced vegetative growth (total vegetative mass). Elevated CO2 stimulated leaf and root growth in both varieties. Water availability diminished negative effects of high temperature on vegetative growth. Yield response to climate-change related factors was year-depended, however it was affected by drought each year., Authors acknowledge INNOVINE European project (No 311775), Aragón Government (A03 group) and “Ministerio de Ciencia e Innovación” of Spain (MCINN AGL2014-56075-C2- 1-R) for funding and “Asociación de Amigos de la Universidad de Navarra” for T. Kizildeniz grant.

Trabajo presentado en el International Congress of Grapevine and Wine Sciences, celebrado en Logroño (España), del 7 al 9 de noviembre de 2018

10 Pags.- 4 Figs.- 3 Tabls., In the Mediterranean area, climate change is associated with atmospheric CO2 concentration increases, enhanced temperatures and scarce water availability, limiting seriously crop yield and decreasing quality. The aim of this study was to investigate the effects of elevated CO2, elevated temperature and water deficit, acting individually and/or interacting, on vegetative and reproductive growth, substrate and plant water status, and must quality in fruit-bearing cuttings of two grapevine (Vitis vinifera L.) cultivars (red and white Tempranillo). In four temperature gradient greenhouses, eight treatments were applied, from fruit set to maturity: CO2 level (400 versus 700 μmol mol−1), temperature (ambient versus ambient +4 °C), and water availability (full irrigation versus cyclic drought). Effects of climate change on grape yield and quality were cultivar dependent. Generally, red Tempranillo had more vegetative growth and grape yield than the white cultivar. Also, grape yield was less affected by the treatments than vegetative growth. Drought, especially under elevated temperature, drastically reduced vegetative growth, bunch fresh and dry weights in both cultivars. Interestingly, elevated CO2 attenuated these negative effects of drought. The effects of climatic factors on yield were not associated with a worse water status of the vegetative or reproductive organs. In red Tempranillo, the combination of elevated CO2, elevated temperature and drought reduced total polyphenol index (TPI), malic acid and increased color density, but did not modify anthocyanin concentration. In white Tempranillo, the combined action of the three factors associated with climate change modified only tartaric acid. In this latter cultivar, drought increased TPI under ambient temperature, regardless of CO2 level, when compared with full-irrigated plants. In conclusion, climate change-related factors (elevated CO2, elevated temperature and water deficit) individually (especially drought) and/or interacting affected to different extent red and white Tempranillo vegetative growth and yield. Drought combined with elevated temperatures reduced grapevine performance, and elevated CO2 mitigated such deleterious effect., Authors thanks Asociación de Amigos de la Universidad de Navarra (T. Kizildeniz PhD Grant), IAMZ-CIHEAM (I. Mekni Master Grant), European project (INNOVINE Call FP7-KBBE-2011-6, Proposal N° 311775), Spanish Ministry of Economy and Innovation (MINECO BFU2011-26989) and Aragón Government (A03 Research Group) for financial support.

46 Pags.- 4 Figs.- 4 Tabls., Although plant performance under elevated CO2 has been extensively studied in the past little is known about photosynthetic performance changing simultaneously CO2, water availability and temperature conditions. Moreover, despite of its relevancy in crop responsiveness to elevated CO2 conditions, plant level C balance is a topic that, comparatively, has received little attention. In order to test responsiveness of grapevine photosynthetic apparatus to predicted climate change conditions, grapevine (Vitis vinifera L. cv. Tempranillo) fruit-bearing cuttings were exposed to different CO2 (elevated, 700 ppm versus ambient, ca. 400 ppm), temperature (ambient versus elevated, ambient +4 °C) and irrigation levels (partial versus full irrigation). Carbon balance was followed monitoring net photosynthesis (AN, C gain), respiration (RD) and photorespiration (RL) (C losses). Modification of environment 13C isotopic composition (δ13C) under elevated CO2 (from -10.30 to -24.93 ‰) enabled the further characterization of C partitioning into roots, cuttings, shoots, petioles, leaves, rachides and berries. Irrespective of irrigation level and temperature, exposure to elevated CO2 induced photosynthetic acclimation of plants. C/N imbalance reflected the inability of plants grown at 700 ppm CO2 to develop strong C sinks. Partitioning of labeled C to storage organs (main stem and roots) did not avoid accumulation of labeled photoassimilates in leaves, affecting negatively Rubisco carboxylation activity. The study also revealed that, after 20 days of treatment, no oxidative damage to chlorophylls or carotenoids was observed, suggesting a protective role of CO2 either at current or elevated temperatures against the adverse effect of water stress., We thank Spanish Ministry of Science and Innovation (BFU2008-01405/BFI), Fundación Universitaria de Navarra (Plan de Investigación de la Universidad de Navarra), Caja Navarra and Gobierno de Aragón (A03 research group) for financial support, Asociación de Amigos de la Universidad de Navarra for Carolina Salazar-Parra grant. Iker Aranjuelo was the recipient of a “Ramon y Cajal” research grant funded by the Spanish Economy and Competitiveness Ministry.

14 Pags.- 1 Tabl.- 6 Figs., Foreseen climate change is expected to impact on grape composition, both sugar and pigment content. We tested the hypothesis that interactions between main factors associated with climate change (elevated CO2, elevated temperature, and water deficit) decouple sugars and anthocyanins, and explored the possible involvement of vegetative area, photosynthesis, and grape C uploading on the decoupling. Tempranillo grapevine fruit-bearing cuttings were exposed to CO2 (700 vs. 400 ppm), temperature (ambient vs. + 4 °C), and irrigation levels (partial vs. full) in temperature-gradient greenhouses. In a search for mechanistic insights into the underlying processes, experiments 1 and 2 were designed to maximize photosynthesis and enlarge leaf area range among treatments, whereas plant growth was manipulated in order to deliberately down-regulate photosynthesis and control vegetative area in experiments 3 and 4. Towards this aim, treatments were applied either from fruit set to maturity with free vegetation and fully irrigated or at 5–8% of pot capacity (experiments 1 and 2), or from veraison to maturity with controlled vegetation and fully irrigated or at 40% of pot capacity (experiments 3 and 4). Modification of air 13C isotopic composition under elevated CO2 enabled the further characterization of whole C fixation period and C partitioning to grapes. Increases of the grape sugars-to-anthocyanins ratio were highly and positively correlated with photosynthesis and grape 13C labeling, but not with vegetative area. Evidence is presented for photosynthesis, from fruit set to veraison, and grape C uploading, from veraison to maturity, as key processes involved in the establishment and development, respectively, of the grape sugars to anthocyanins decoupling., Science and Innovation (BFU2008-01405/BFI), Economy and Competitiveness (AGL2014-56075-C2-1-R) Spanish Ministries, Aragón Government (A03 group) and the Basque Government (IT-932-16) for financial support, Navarra University “Asociación de Amigos” for Carolina Salazar-Parra grant

[Aim]: To test the role of mountains as barriers to gene flow in co-distributed taxa with different life history traits. [Location]: Sierra de Guadarrama, Central Spain. [Methods]: We used larval genotypes of four amphibian species (Epidalea calamita, Hyla molleri, Pelophylax perezi and Pelobates cultripes) sampled on northern and southern slopes of Sierra de Guadarrama to describe genetic structure with F, migration rates per generation, clustering algorithms and resistance by elevation surfaces. We also recorded individual displacement events as a proxy of dispersal potential during a seven-year monitoring project based on capture–mark–recapture (CMR). [Results]: All species travelled longer cumulative distances than those reported in the study area for P. cultripes (0.71 km). Individuals of E. calamita travelled up to 3.55 km, followed by H. molleri (2.84 km) and P. perezi (1.51 km). Pairwise F estimates showed lower overall connectivity in P. cultripes. Average migration rates per generation were low in all species, with exceptions in same-slope populations of H. molleri and P. cultripes. Clustering algorithms consistently recovered well-differentiated population groups of P. cultripes in northern versus southern slopes, but widely admixed areas were observed in the other species, especially near mountain passes. Resistance by elevation surfaces showed a strong barrier effect of Sierra de Guadarrama in P. cultripes and suggested a potential role of topography in the genetic structure of E. calamita and H. molleri. [Main conclusions]: Sierra de Guadarrama currently acts as a strong barrier to gene flow for P. cultripes and, to a lesser extent, for E. calamita, H. molleri and P. perezi. This differential effect can be partly explained by differences in life history traits, including dispersal potential. Our findings support the general role of the Central System as a key feature shaping population connectivity and genetic variation in amphibian communities., G.S.M. was funded by Asociación de Amigos de la Universidad de Navarra. This research was funded by grants CGL2008-04271-C02-01/BOS and CGL2011-28300 (Ministerio de Ciencia e Innovación, Ministerio de Economía y Competitividad, Spain, and FEDER) toI.M.S., who was supported by funding from the Spanish ‘Ramón yCajal’(RYC-2007-01668) and ‘Severo Ochoa’ (SEV-2012-0262) programmes

Leguminous plants grown in sewage sludge-amended soils can acquire nitrogen by assimilation of nitrate and ammonium from the soil solution or from atmospheric-dinitrogen (N2) fixation through association with N2-fixing bacteria. We proposed that operation of both metabolic processes could contribute to alleviate the impact of drought in sludge-treated plants. A greenhouse experiment was conducted to evaluate the involvement of nodule metabolism in the use efficiency of water and N in sludge-treated plants. Treatments comprised (1) plants inoculated with rhizobia and amended with sewage sludge; (2) plants inoculated with rhizobia without any amendment; and (3) noninoculated plants supplied with ammonium nitrate, each under well-watered and drought conditions. Under drought, sludge-treated plants had increased plant growth and higher photosynthetic and water-use efficiencies than untreated plants. Drought stimulated nitrate reductase and GS/GOGAT activities but did not affect the activities of phosphoenolpyruvate carboxylase and malate dehydrogenase or the leghemoglobin concentration. The results suggest that under drought conditions, both N2 fixation and nitrate assimilation in nodules of sludge-treated plants contributed to improve plant N supply and to increase the drought tolerance of alfalfa. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim., Authors thank NILSA (Navarra de Infraestructuras Locales, S.A.) for providing sewage sludge and sludge analysis, A. Urdiáin and M. Oyárzun for technical assistance during measurements. M. L. Fiasconaro was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

Efforts to mitigate amphibian declines are hindered by a lack of information about basic aspects of their biology and demography. The effective to census population size ratio (N/N) is one of the most important parameters for the management of wildlife populations because it combines information on population abundance and genetic diversity and helps predict population viability in the long term. Few studies have calculated this ratio in amphibians, which sometimes show low ratios, associated with a higher extinction risk. Here we integrate field-based (capture-mark-recapture studies, egg string counts) and molecular approaches (estimation of the effective number of breeders (N) and the effective population size (N) based on genotypes from larval cohorts and candidate parents) to produce the first estimates of the N/N and N/N ratios in two amphibians, the Iberian ribbed newt Pleurodeles waltl and the western spadefoot Pelobates cultripes. Additionally, we investigate sex-biased dispersal in both species based on direct (field observations) and indirect (genetic) evidence. Both species showed similar ratios, slightly lower in Pleurodeles (0.21–0.24) than in Pelobates (0.25–0.30). Observed displacement rates were low in both species (P. waltl = 0.51%; P. cultripes = 1.23%). We found no evidence for sex-biased dispersal in P. cultripes, but both direct and indirect evidences suggest a tendency for female-biased dispersal in P. waltl. We discuss differences in the genetic estimates of N and N provided by three inference methods and the implications of our findings for the management of these species, characteristic of Mediterranean wetlands in the Iberian Peninsula and listed as Near Threatened., JGR was supported by the Consejo Superior de Investigaciones Científicas of Spain (CSIC) and the European Social Fund (ESF) (JAE-pre PhD fellowship). GSM is funded by a predoctoral grant provided by the Asociación de Amigos de la Universidad de Navarra. This research was funded by grants CGL2008-04271-C02-01/BOS, and CGL2011-28300 (Ministerio de Ciencia e Innovación -MICINN-), Ministerio de Economía y Competitividad -MEC-, Spain, and FEDER) to IMS, who was supported by funding from the Spanish “Ramón y Cajal” and Severo Ochoa (SEV-2012-0262) programs.

In recent decades, mean global temperatures have increased in parallel with a sharp rise in atmospheric carbon dioxide (CO2) levels, with apparent implications for precipitation patterns. The aim of the present work is to assess the sensitivity of different phenological stages of grapevine to temperature and to study the influence of other factors related to climate change (water availability and CO2 concentration) on this relationship. Grapevine phenological records from 9 plantings between 42.75°N and 46.03°N consisting of dates for budburst, flowering and fruit maturity were used. In addition, we used phenological data collected from 2 years of experiments with grapevine fruit-bearing cuttings with two grapevine varieties under two levels of water availability, two temperature regimes and two levels of CO2. Dormancy breaking and flowering were strongly dependent on spring temperature, while neither variation in temperature during the chilling period nor precipitation significantly affected budburst date. The time needed to reach fruit maturity diminished with increasing temperature and decreasing precipitation. Experiments under semi-controlled conditions revealed great sensitivity of berry development to both temperature and CO2. Water availability had significant interactions with both temperature and CO2; however, in general, water deficit delayed maturity when combined with other factors. Sensitivities to temperature and CO2 varied widely, but higher sensitivities appeared in the coolest year, particularly for the late ripening variety, ‘White Tempranillo’. The knowledge gained in whole plant physiology and multi stress approaches is crucial to predict the effects of climate change and to design mitigation and adaptation strategies allowing viticulture to cope with climate change., This work was funded by the Land Settlement Association, the University of Reading Research Endowment Trust and East Malling Trust, European Union (INNOVINE Call FP7-KBBE-2011-6, Proposal N°311775), Ministerio de Ciencia e Innovación of Spain (MCINN BFU 2011-26989), Ministerio de Economía y Competitividad of Spain (AGL2014-56075-C2-1-R), Aragón Government (A03 research group) and Asociación de Amigos de la Universidad de Navarra (grant to TK).

Elevated CO 2 may decrease alfalfa forage quality and in vitro digestibility through a drop in crude protein and an enhancement of fibre content. The aim of the present study was to analyse the effect of elevated CO 2, elevated temperature and Sinorhizobium meliloti strains (102F78, 102F34 and 1032 GMI) on alfalfa yield, forage quality and in vitro dry matter digestibility. This objective is in line with the selection of S. meliloti strains in order to maintain high forage yield and quality under future climate conditions. Plants inoculated with the 102F34 strain showed more DM production than those inoculated with 1032GMI; however, these strains did not show significant differences with 102F78 plants. Neutral or acid detergent fibres were not enhanced in plants inoculated with the 102F34 strain under elevated CO 2 or temperature and hence, in vitro dry matter digestibility was unaffected. Crude protein content, an indicator of forage quality, was negatively related to shoot yield. Plants inoculated with 102F78 showed a similar shoot yield to those inoculated with 102F34, but had higher crude protein content at elevated CO 2 and temperature. Under these climate change conditions, 102F78 inoculated plants produced higher quality forage. However, the higher digestibility of plants inoculated with the 102F34 strain under any CO 2 or temperature conditions makes them more suitable for growing under climate change conditions. In general, elevated CO 2 in combination with high temperature (Climate Change scenario) reduced IVDMD and CP content and enhanced fibre content, which means that animal production will be negatively affected. © 2012 Elsevier GmbH., This work was supported by Ministerio de Ciencia e Innovación (MICINN BFU2008-01405 and BFU2011-26989), Fundación Universitaria de Navarra (PIUNA-2008) and Fundación Caja Navarra. A. Sanz-Sáez was the recipient of a research grant from Asociación de Amigos de la Universidad de Navarra. The assistance of A. Urdiain, M. Oyarzun and L. González is also appreciated.

The aim of the present study was to analyse the effect of elevated CO 2 at different temperature, inoculation with three Sinorhizobium meliloti strains and growth in two seasons (summer and autumn) on plant growth, photosynthesis and N 2 fixation (estimated as apparent nitrogenase activity, ANA). One of the objectives was to demonstrate that most efficient N 2-fixing strains could improve alfalfa forage production. The second objective was to test if S. meliloti strains alter their efficiency under climate change conditions. The third objective was to show if seasonal changes may modify alfalfa productivity. Interaction of CO 2 and temperature enhanced alfalfa dry matter in both seasons; however, plants produced more dry matter in autumn than in summer, due to the negative effect of elevated summer temperature on N 2-fixation. Higher yield in summer corresponded to plants in symbiosis with 102F78 strain being not related to enhanced nodule dry matter or apparent nitrogenase activity but to putative lower carbon consumption for N 2 fixing process. Contrariwise, in autumn the highest yield was obtained by 102F34 as a consequence of increased nodule dry matter induced under elevated CO 2 and temperature. © 2011 Elsevier B.V., This work was supported by Ministerio de Ciencia e Innovación (MICINN BFU2008-01405), Fundación Universitaria de Navarra (PIUNA-2008) and Fundación Caja Navarra. A. Sanz-Sáez was the recipient of a research grant from Asociación de Amigos de la Universidad de Navarra. The assistance of A. Urdiain, M. Oyarzun and L. González is also appreciated.

Sewage sludge has been used as N fertilizer because it contains some of inorganic N, principally as nitrate and ammonium ions. However, sewage sludge addition to legumes could result in impaired nodule metabolism due to the presence of inorganic N from sludge. A greenhouse experiment was conducted to examine the effects of sewage sludge on growth, photosynthesis, nitrogen assimilation and nodule metabolism in alfalfa (Medicago sativa L. cv. Aragón). Plants were grown in pots with a mixture of perlite and vermiculite (2:1, v/v). The experiment included three treatments: (1) plants inoculated with rhizobia and amended with sewage sludge at rate of 10% (w/w) (RS); (2) plants inoculated with rhizobia without any amendment (R); and (3) non-inoculated plants fed with ammonium nitrate (N). N2-fixing plants had lower growth and sucrose phosphate synthase activity but higher photosynthesis than nitrate-fed plants because they compensated the carbon cost of the rhizobia. However, sewage sludge-treated plants evidenced a loss of carbon sink strength due to N2 fixation by means of decreased photosynthetic capacity, leaf chlorophylls and N concentration in comparison to untreated plants. Sewage sludge did no affect nodulation but decreased nodule enzyme activities involved in carbon and N metabolisms that may lead to accumulation of toxic N-compounds., This project was supported by NILSA. M.L. Fiasconaro was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

Beech forests naturally regenerating from clear-cutting can exhibit different microclimates depending on size of saplings and stem density. When beech trees are young and stem density is low, the level of radiation inside the ecosystem reaching the soil surface is high; consequently, air and soil temperatures rise and the soil water content may decrease. These microclimatic parameters presumably will affect the anatomy, photosynthesis, and carbon metabolism of beech leaves. We studied the morphology and physiology of sun and shade leaves of beech trees differing in age and growing within clear-cut areas with distinct microclimate. Results were compared with those of adult trees in an unmanaged forest. We selected a stand clear-cut in 2001 (14,000 trees ha−1), another clear-cut in 1996 (44,000 trees ha−1) and an unmanaged forest (1,000 trees ha−1). Photosynthetic photon flux density (PPFD) incident on sun leaves, air temperature, soil moisture, and soil temperature within the forests affected water status and carbohydrate storage in all trees. As trees became older, PPFD also influenced pigment composition and Rubisco activity in sun leaves. On the other hand, shade leaves from the oldest trees were the most sensitive to PPFD, air temperature, and soil moisture and temperature inside the forest. Contrariwise, microclimatic parameters slightly affected the physiology of shade leaves of the beech in the stand with the highest light attenuation. Air and soil temperatures were the parameters that most affected the photosynthetic pigments and carbohydrate storage in shade leaves of the youngest trees., Our research was supported by Fundación Universitaria de Navarra (FUNA, PIUNA) and Caja Navarra. Iván Closa was a recipient of a grant from Asociación de Amigos de la Universidad de Navarra (ADA).

Trabajo presentado a las II Jornadas del Grupo de Viticultura y Enología (SECH), celebradas en Madrid el 3 y 4 de noviembre de 2016., El calentamiento global proyectado puede afectar de forma negativa al proceso de maduración de la uva y a su composición. En este sentido, el empleo de diferentes variantes somáticas que se acumulan en cada variedad en el proceso de multiplicación vegetativa de la vid (Vitis vinifera L.) pueden ser una herramienta útil para mantener la calidad de la uva y el vino en condiciones de cambio climático. El objetivo de este trabajo fue evaluar el efecto de la temperatura elevada sobre la acumulación de azúcares y antocianinas en las bayas de trece variantes somáticas (accesiones) de vid cv. Tempranillo, que difieren en la longitud de su ciclo. Se cultivaron estaquillas fructíferas de trece accesiones de vid (86, 1052, 336, 518, 501, 349, 280, 825, 807, 814, 318, 56, y 1084, Gobierno de la Rioja) en condiciones controladas, y se sometieron, desde cuajado hasta madurez, a dos regímenes térmicos: 24/14ºC o 28/18ºC, día/noche. Se determinó el desarrollo fenológico como el número de días desde cuajado a envero y desde envero a madurez. Se analizó la concentración de sólidos solubles totales (SST) y antocianinas totales en piel en cuatro puntos del desarrollo de la baya: i) pre-envero (antes del inicio de la síntesis de antocianinas), ii) envero (50 % de la baya coloreada), iii) dos semanas después del envero y iv) madurez (21-23ºBrix). Las accesiones 349, 807, 814, 56 y 1084 presentaron los ciclos más largos y la 501 y 518 los más cortos. Las principales diferencias entre accesiones se observaron en el periodo comprendido entre envero y madurez. La temperatura elevada redujo significativamente el número de días transcurridos entre cuajado y madurez, con una mayor influencia antes del envero. Las variantes somáticas difirieron en la tasa de acumulación de SST y antocianinas. Los resultados indican, en general, un efecto de desacoplamiento térmico entre azúcares y antocianinas en la baya. Las variantes somáticas estudiadas difirieron en el grado de desacoplamiento entre ambos metabolitos, así las accesiones 501 y 56 fueron las que mostraron un mayor desequilibrio, especialmente en madurez, mientras que la 1052 y la 318 fueron las menos afectadas. Estas diferencias no estuvieron asociadas directamente con las distintas longitudes de ciclo (cuajadoenvero) que mostraron las accesiones estudiadas., Ministerio de Ciencia e Innovación (MCINN AGL2014-56075-C2-1-R), Unión Europea (INNOVINE Call FP7-KBBE-2011-6, Proposal Nº 311775), Gobierno de Aragón Grupo de investigación A03) y Asociación de Amigos de la Universidad de Navarra (Beca M.A).

9 Pags.- 3 Tabls.- 4 Figs., The increase in grape berry ripening rates associated to climate change is a growing concern for wine makers as it rises the alcohol content of the wine. The present work studied the combined effects of elevated CO2, temperature and UV-B radiation on leaf physiology and berry ripening rates. Three doses of UV-B: 0, 5.98, 9.66 kJ m−2 d−1, and two CO2–temperature regimes: ambient CO2-24/14 °C (day/night) (current situation) and 700 ppm CO2-28/18 °C (climate change) were imposed to grapevine fruit-bearing cuttings from fruit set to maturity under greenhouse-controlled conditions. Photosynthetic performance was always higher under climate change conditions. High levels of UV-B radiation down regulated carbon fixation rates. A transient recovery took place at veraison, through the accumulation of flavonols and the increase of antioxidant enzyme activities. Interacting effects between UV-B and CO2–temperature regimes were observed for the lipid peroxidation, which suggests that UV-B may contribute to palliate the signs of oxidative damage induced under elevated CO2–temperature. Photosynthetic and ripening rates were correlated. Thereby, the hastening effect of climate change conditions on ripening, associated to higher rates of carbon fixation, was attenuated by UV-B radiation., This work was supported by Ministerio de Ciencia e Innovación of Spain (MCINNBFU2011-26989), INNOVINE Call FP7-KBBE-2011-6, Proposal N° 311775, Asociación de Amigos de la Universidad de Navarra (grant to J. M-L) and UV4growth COST Action FA0906.

34 Pags.- 3 Tabls.- 6 Figs. The definitive version is available at: http://www.sciencedirect.com/science/journal/01689452, This work aims to characterize the physiological response of grapevine (Vitis vinifera L.) cv. Tempranillo to UV-B radiation under water deficit conditions. Grapevine fruit-bearing cuttings were exposed to three levels of supplemental biologically effective UV-B radiation (0, 5.98 and 9.66 kJ m−2 day−1) and two water regimes (well watered and water deficit), in a factorial design, from fruit-set to maturity under glasshouse-controlled conditions. UV-B induced a transient decrease in net photosynthesis (Anet), actual and maximum potential efficiency of photosystem II, particularly on well watered plants. Methanol extractable UV-B absorbing compounds (MEUVAC) concentration and superoxide dismutase activity increased with UV-B. Water deficit effected decrease in Anet and stomatal conductance, and did not change non-photochemical quenching and the de-epoxidation state of xanthophylls, dark respiration and photorespiration being alternative ways to dissipate the excess of energy. Little interactive effects between UV-B and drought were detected on photosynthesis performance, where the impact of UV-B was overshadowed by the effects of water deficit. Grape berry ripening was strongly delayed when UV-B and water deficit were applied in combination. In summary, deficit irrigation did not modify the adaptive response of grapevine to UV-B, through the accumulation of MEUVAC. However, combined treatments caused additive effects on berry ripening., This work was supported by Fundación Universitaria de Navarra, Navarra-Aquitaine Cooperation Program (Gobierno de Navarra and Conseil Régional d’Aquitaine), Ministerio de Ciencia e Innovación of Spain (MCINN BFU2011-26989), European Union (FP7-KBBE-2012-6-singlestage), Asociación de Amigos de la Universidad de Navarra (grant to J. M-L), UV4growth COST Action FA0906 and Gobierno de Aragón (A03 research group).

[Background and Aims] The application of deficit irrigation to grapevines modifies the hormonal status of berries, but little information about the influence of berry hormones on phenological sensitivity to water deficit is available. Therefore, the aim of this research was to assess the involvement of berry hormonal status in fruit composition in response to regulated deficit irrigation applied during different phenological stages in two grapevine cultivars., [Methods and Results] The study was carried out on fruiting cuttings of two cultivars of Vitis viniferaL., Tempranillo and Graciano. Treatments were: (i) early water deficit from fruitset to onset of veraison (early deficit); (ii) late water deficit from onset of veraison to harvest (late deficit); and (iii) plants regularly irrigated (Control). Both early water deficit and late water-deficit strategies modified evolution of indole-3-acetic acid, abscisic acid, salicylic acid and jasmonic acid, which was related to changes in berry size, increases in phenolic substances and accumulation of amines., [Conclusions] Differential sensitivity of Tempranillo and Graciano grapevines to seasonal water stress was mediated, at least in part, by alterations in hormonal status of berries at the time of water stress imposition., [Significance of Study] This study relates interspecific differences in the sensitivity of seasonal water-deficit irrigation to changes in the endogenous hormonal status of berries. © 2014 Australian Society of Viticulture and Oenology Inc., This project was supported by the Ministerio de Ciencia e Innovación (MCINN BFU2011-26989) of Spain. Maria Niculcea was the recipient of a grant from the Asociación de Amigos de la Universidad de Navarra.

The association of Lactuca sativa L. with arbuscular mycorrhizal fungi (AMF) can benefit growth and induce the accumulation of mineral nutrients and antioxidants in leaves. Our objectives were (1) to assess to what extent growing season influence growth and nutritional quality of the green-leaf Batavia Rubia Munguía (BRM) and the red-leaf Maravilla de Verano (MV) lettuce cultivars; (2) to test if the growing season affected the effectiveness of AMF as enhancers of the nutritional quality of the aforementioned cultivars; and (3) to check if inoculation with AMF allows extending the season to cultivate these two lettuce cultivars, without a loss of plant growth or nutritional quality. Results showed that winter was the most favourable growing season for both types of lettuce but mycorrhizal plants cultivated in other seasons different from winter achieved higher or similar shoot biomass than their respective non-inoculated controls in winter. AMF induced the accumulation of Fe, proteins, carotenoids and anthocyanins in both cultivars of lettuce in winter and spring. In summer and autumn, both types of lettuce showed increased levels of anthocyanins when associated with AMF. Mycorrhizal BRM also had enhanced levels of K, sugars and ascorbate in winter and spring as well as increased amounts of Mg and Cu in winter and summer. Mycorrhizal MV had increased quantities of Cu, Zn and sugars in spring and Mn in autumn. After estimating intakes of mineral elements through the consumption of 150. g per day of fresh leaves from mycorrhizal lettuces we may affirm that there would not be toxicological risk according to the Recommended Daily Allowances (RDAs). The application of AMF could allow extending cultivation of lettuce to other periods not appropriate for producing non-mycorrhizal plants. © 2013 Elsevier B.V., Marouane Baslam received a grant from Asociación de Amigos de la Universidad de Navarra (ADA).

Available online 24 May 2014, Continued emissions of CO2, derived from human activities, increase atmospheric CO2 concentration. The CO2 rise stimulates plant growth and affects yield quality. Effects of elevated CO2 on legume quality depend on interactions with N2-fixing bacteria and mycorrhizal fungi. Growth at elevated CO2 increases photosynthesis under short-term exposures in C3 species. Under long-term exposures, however, plants generally acclimate to elevated CO2 decreasing their photosynthetic capacity. An updated survey of the literature indicates that a key factor, perhaps the most important, that characteristically influences this phenomenon, its occurrence and extent, is the plant source–sink balance. In legumes, the ability of exchanging C for N at nodule level with the N2-fixing symbionts creates an extra C sink that avoids the occurrence of photosynthetic acclimation. Arbuscular mycorrhizal fungi colonizing roots may also result in increased C sink, preventing photosynthetic acclimation. Defoliation (Anthyllis vulneraria, simulated grazing) or shoot cutting (alfalfa, usual management as forage) largely increases root/shoot ratio. During re-growth at elevated CO2, new shoots growth and nodule respiration function as strong C sinks that counteracts photosynthetic acclimation. In the presence of some limiting factor, the legumes response to elevated CO2 is weakened showing photosynthetic acclimation. This survey has identified limiting factors that include an insufficient N supply from bacterial strains, nutrient-poor soils, low P supply, excess temperature affecting photosynthesis and/or nodule activity, a genetically determined low nodulation capacity, an inability of species or varieties to increase growth (and therefore C sink) at elevated CO2 and a plant phenological state or season when plant growth is stopped., Authors thank Genoma España (within a collaborative agreement with Genome Canada (Grapegen Project)), the Innovine Project (Combining innovation in vineyard management and genetic diversity for a sustainable European viticulture (Call FP7-KBBE-2012-6, Proposal N◦311775-INNOVINE)), the Spanish Ministry of Science and Innovation [grant numbers BFU2008-01405/BFI and BFU2011-26989], Fundación Universitaria de Navarra (Plan de Investigación de la Universidad de Navarra), Caja Navarra and Gobierno de Aragón (A03 research group) for financial support, Asociación de Amigos de la Universidad de Navarra for PhD Thesis grants.

13 Págs., 3 Figs., 4 Tabls., Water-deficit irrigation to grapevines reduces plant growth, yield, and berry growth, altering the ripening process, all of which may influence fruit composition and wine quality. Therefore, the goals of this study were (1) to investigate the influence of the main endogenous berry hormones, abscisic acid (ABA), indole-3-acetic acid (IAA), salicylic acid (SA), and jasmonic acid (JA), on berry growth and ripening under water-deficit conditions and (2) to analyze changes in fruit composition, specifically N compounds, under water deprivation. The study was carried out using container-grown Tempranillo grapevines grown under controlled conditions in a greenhouse. Two irrigation treatments were imposed: control (well-watered) and sustained deficit irrigation (SDI). Water deficit decreased leaf area and the source-to-sink ratio, reduced yield and berry size, and decreased concentrations of the main phenolic compounds. SDI also modified berry hormonal status. At the pea-size stage, SDI berries had lower IAA and higher JA and SA than nonstressed berries. At veraison (onset of ripening), accumulation of ABA was less accentuated in SDI than in control berries. At harvest, the content of amino acids and free ammonium was low in both treatments but SDI-treated berries showed a significant accumulation of amines. Results suggest that water restrictions to grapevines might be playing a physiological role in reducing berry growth through affecting hormone dynamics, phenolic synthesis, and the berry amino acid content and composition, which could compromise fruit quality. Possible roles of endogenous IAA controlling berry size and endogenous ABA and SA controlling levels of anthocyanins and flavonols at harvest are discussed. © 2013 Springer Science+Business Media New York., This work was supported by Fundación Universitaria de Navarra (2011) and Ministerio de Ciencia e Innovación (MCINN BFU2011-26989), Spain. F. Morales thanks Gobierno de Aragón (A03 research group) for financial support. M. Niculcea was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

9 Pags., 6 Figs., 2 Tabls., Lettuce (Lactuca sativa L.) is a major food crop within the European Union. The objective of this study was to test the potential nutritional quality of three types of lettuce consumed as salads: Lactuca sativa L. var. longifolia (commonly named Cogollos de Tudela) and two cultivars of Lactuca sativa L. var. capitata (Batavia Rubia Munguía and Maravilla de Verano). Distribution of sugars, proteins, minerals and antioxidant compounds (carotenoids, chlorophylls, phenolics, anthocyanins and ascorbate) between outer and inner leaves was tested. The potential benefits of each variety or cultivar were due to different compounds accumulated in leaves. Cogollos de Tudela showed higher levels of Mg and Ca in both external and internal leaves, and greater concentrations of Mn, chlorophylls and carotenoids in inner leaves than lettuces belonging to the var. capitata. Batavia Rubia Munguía had low quantity of Na and an important amount of K, Mg, Ca, Fe and Zn in outer leaves and it was the cultivar that accumulated the highest quantity of water in both outer and inner leaves. Maravilla de Verano showed the greatest contents of anthocyanins and ascorbate in external leaves. In the three cultivars some of the potentially beneficial compounds appeared in higher levels in the outer than in the inner leaves. Therefore, the complete elimination of the external leaves should be reconsidered when lettuces are consumed as salads or used as food crop for the ‘Fourth Range’ of vegetables., Marouane Baslam is the recipient of a grant from Asociación de Amigos de la Universidad de Navarra (ADA). Fermín Morales thanks Gobierno de Aragón (A03 research group) for financial support.

Trabajo presentado en el XIII Congresso Luso-Espanhol de Fisiologia Vegetal, celebrado en Lisboa del 24 al 28 de julio de 2013., Experiments were conducted in grapevine (Vitis vinifera L. cv. Tempranillo) fruit-bearing cuttings, using temperature gradient greenhouses (TGG), to investigate the possible effects of future climate scenarios: elevated CO2 (700 ppm) vs. ambient (ca.375 ppm), elevated temperature (+4ºC) vs. ambient and partia l vs. full irrigation, on C balance, partitioning and grape quality. Carbon balance was followed monitoring net photosynthesis (AN, C gain), respiration (RD) and photorespiration (RL) (C losses). For C partitioning characterization, 13 C isotopic composition (δ13C) was modified (-24.93 ‰) in the elevated CO2 TGGs, with respect to that of the ambient TGGs (-10.30 ‰). Elevated CO2 increased transiently (after 10 days of treatment) AN irrespective of water availability and temperature, but effects disappeared 10 days later. In most cases, no differences in RL were found. When grown under ambient temperature (regardless of water status), leaves respired more under elevated than under current CO2 concentrations at both sampling dates, whereas all leaves respired similarly to controls when grown under elevated temperature. Carbon and N isotopic composition (δ13C and δ15N respectively) was characterized in roots, cuttings, shoots, petioles, leaves, rachides and berries. Data revealed a low C-sink strength of cuttings and rachides, whereas leaf N decreased as a response to elevated CO2 . Plants exposed to elevated CO2 did acclimate. Such acclimation was reflected in lower AN rates measured at elevated CO2 and a higher C/N ratio in plants grown at elevated CO2. After 20 days of treatment, no oxidative damage to chlorophylls or carotenoids was observed, suggesting a protective role of CO2 either at current or elevated temperatures against the adverse effect of water stress. Treatments simulating climate change influenced g rapevine maturation. Climate change (elevated CO2 in interaction with water deficit, but irrespective of temperature) shortened maturation time. pH and extractable anthocyanins were not affected, whereas acidity, malic acid, total and potential anthocyanins, phenolic richness, colour intensity and tonality index were only slightly altered under some particular circumstances., Project funding by Spanish Ministry of Economy and Competitiveness [grant No. BFU2008-01405/BFI, BFU2011-26989 and AGL2011-30386-CO2-O2], Fundación Universitaria de Navarra (Plan de Investigación de la Universidad de Navarra), Caja Navarra, Gobierno de Aragón (A03 research group). Fellowships: Asociación de Amigos de la Universidad de Navarra to C S -P. Special thanks to A. Urdiain and M. Oyarzun for their technical assistance.

Lettuce, a major food crop within the European Union and the most used for the so-called 'Fourth Range' of vegetables, can associate with arbuscular mycorrhizal fungi (AMF). Mycorrhizal symbiosis can stimulate the synthesis of secondary metabolites, which may increase plant tolerance to stresses and enhance the accumulation of antioxidant compounds potentially beneficial to human health. Our objectives were to assess (1) if the application of a commercial formulation of AMF benefited growth of lettuce under different types and degrees of water deficits; (2) if water restrictions affected the nutritional quality of lettuce; and (3) if AMF improved the quality of lettuce when plants grew under reduced irrigation. Two cultivars of lettuce consumed as salads, Batavia Rubia Munguía and Maravilla de Verano, were used in the study. Four different water regimes were applied to both non-mycorrhizal and mycorrhizal plants: optimal irrigation (field capacity [FC]), a water regime equivalent to 2/3 of FC, a water regime equivalent to 1/2 of FC and a cyclic drought (CD). Results showed that mycorrhizal symbiosis improved the accumulation of antioxidant compounds, mainly carotenoids and anthocyanins, and to a lesser extent chlorophylls and phenolics, in leaves of lettuce. These enhancements were higher under water deficit than under optimal irrigation. Moreover, shoot biomass in mycorrhizal lettuces subjected to 2/3 of FC were similar to those of non-mycorrhizal plants cultivated under well-watered conditions. In addition, lettuces subjected to 2/3 FC had similar leaf RWC than their respective well-watered controls, regardless of mycorrhizal inoculation. Therefore, results suggest that mycorrhizal symbiosis can improve quality of lettuce and may allow restrict irrigation without reducing production. © 2011 Springer-Verlag., Marouane Baslam is the recipient of a grant from Asociación de Amigos de la Universidad de Navarra (ADA).

Photosynthetic carbon fixation (A N) and photosynthetic electron transport rate (ETR) are affected by different environmental stress factors, such as those associated with climate change. Under stress conditions, it can be generated an electron excess that cannot be consumed, which can react with O 2, producing reactive oxygen species. This work was aimed to evaluate the influence of climate change (elevated CO 2, elevated temperature and moderate drought) on the antioxidant status of grapevine (Vitis vinifera) cv. Tempranillo leaves, from veraison to ripeness. The lowest ratios between electrons generated (ETR) and consumed (A N + respiration + photorespiration) were observed in plants treated with elevated CO 2 and elevated temperature. In partially irrigated plants under current ambient conditions, electrons not consumed seemed to be diverted to alternative ways. Oxidative damage to chlorophylls and carotenoids was not observed. However, these plants had increases in thiobarbituric acid reacting substances, an indication of lipid peroxidation. These increases matched well with an early rise of H 2O 2 and antioxidant enzyme activities, superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and catalase (EC 1.11.1.6). Enzymatic activities were maintained high until ripeness. In conclusion, plants grown under current ambient conditions and moderate drought were less efficient to cope with oxidative damage than well-irrigated plants, and more interestingly, plants grown under moderate drought but treated with elevated CO 2 and elevated temperature were not affected by oxidative damage, mainly because of higher rates of electrons consumed in photosynthetic carbon fixation. © Physiologia Plantarum 2011., We thank the Spanish Ministry of Science and Innovation (BFU2008-01405/BFI), Fundacion Universitaria de Navarra (Plan de Investigaci ´ on´ de la Universidad de Navarra), Caja Navarra and Gobierno de Aragon (A03 research group) for financial support, ´ Asociacion de Amigos de la Universidad de Navarra for ´ Carolina Salazar-Parra grant, A. Urdiain, M. Oyarzun (University of Navarra) and A. Calvino (Estaci ˜ on Experimental de ´ Aula Dei-CSIC) for excellent technical assistance, and Station of Viticulture and Enology of Navarra (Olite, Navarra, Spain) for dormant cuttings supply

13 Pag., 9 Fig., Greenhouse experiments were conducted to investigate the impact of predicted climate change (elevated CO2, 700 μmol CO2 mol−1 air vs. ambient; elevated temperature, 28/18°C vs. 24/14°C, day/night; and partial irrigation, 40% of field capacity vs. well-irrigated) on grape berry quality characteristics during ripening. Grapevine (Vitis vinifera L. cv. Tempranillo) fruiting cuttings were used as experimental plant material. Climate change shortened the time between grape veraison and full maturity. At harvest time, many of the grape quality parameters determined were affected by the different grape maturity. The data were re-grouped according to total soluble solids to factor out changes due to the shortened time to maturity, and the effects on grape quality were then re-examined. Under current CO2 and temperature conditions, partial irrigation decreased berry malic acid concentration and facilitated anthocyanins extractability. Elevated CO2 and temperature decreased berry malic acid and total anthocyanins potential in well-irrigated plants and increased tonality index, irrespective of water availability. In partial irrigation conditions, elevated CO2 and temperature hindered the anthocyanins extractability. In summary, results indicate that climate change (elevated CO2, high temperature and partial irrigation) affects phenology and berry quality., Financial support from Spanish Ministry of Science and Innovation (BFU2008-01405/BFI), Fundación Universitaria de Navarra (Plan de Investigación de la Universidad de Navarra) and Caja Navarra is gratefully acknowledged. Fermín Morales wishes to thank Gobierno de Aragón (A03 research group) for financial support. Carolina Salazar-Parra was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

The physicochemical and biological properties of soils within an unmanaged beech stand and two stands clear-cut in 2001 or 1996 were studied and compared across the year 2008. The clear-cut stands were left to naturally regenerate and exhibited very different levels of tree density. Soil from the stand clear-cut in 2001 had the lowest contents of organic matter and nitrogen, showed high resistance to penetration and the pH varied throughout the seasons. Basal respiration achieved minimum values in summer in both the unmanaged stand and the stand clear-cut in 1996. However, basal respiration slightly fluctuated from spring to autumn in the stand clear-cut in 2001. The seasonal dynamics of protease and phosphatase activities were similar within the three stands: the maximum protease activity was detected in spring and the highest phophatase activity in winter. β-Glucosidase activity in autumn and dehydrogenase in winter were greater in the unmanaged than in the clear-cut stands. Moreover, dehydrogenase activity was extremely low in the stand clear-cut in 1996. Microclimatic parameters within the stands were significantly correlated with several biological properties of soils, with microclimate being strongly determined by the density of trees. Results also suggested that ectomycorrhizal fungi would be key components of the soil microflora in the beech forests., Our research was supported by Fundación Universitaria de Navarra (FUNA, PIUNA) and Caja Navarra. Iván Closa was a recipient of a grant from the Asociación de Amigos de la Universidad de Navarra (ADA)

17 Pags.- 6 Tabls.- 5 Figs., The projected increase in mean temperatures caused by climate change is expected to have detrimental impacts on berry quality. Microorganisms as arbuscular mycorrhizal fungi (AMF) produce numerous benefits to host plants and can help plants to cope with abiotic stresses such as high temperature. The aims of this research were to characterise the response of three clones of Vitis vinifera L. cv. Tempranillo to elevated temperatures and to determine whether AMF inoculation can improve berry antioxidant properties under these conditions. The study was carried out on three fruit-bearing cuttings clones of cv. Tempranillo (CL-260, CL-1048 and CL-1089) inoculated with AMF or uninoculated and subjected to two temperature regimes (day–night: 24°C-14°C and 28°C-18°C) during berry ripening. Results showed that clonal diversity of Tempranillo resulted in different abilities to respond to elevated temperature and AMF inoculation. In CL-1048, AMF inoculation improved parameters related to phenolic maturity such as anthocyanin content and increased antioxidant activity under elevated temperature, demonstrating a protective role of AMF inoculation against warming effects on berry quality. The results therefore suggest that selection of new clones and/or the implementation of measures to promote the association of grapevines with AMF could be strategies to improve berry antioxidant properties under future warming conditions., This work was supported by Spanish Ministry of Science and Innovation (BFU2011-26989) and INNOVINE European project (INNOVINE 311775). F. Morales thanks Aragón Government (A03 Research Group) for financial support. N. Torres was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

Pelophylax perezi is an Iberian green waterfrog with high tolerance to habitat alteration that at times shows local population growth and demographic expansion, even where other species decline. However, pond destruction, invasive predators and hybridisation with other European waterfrog species (P. ridibundus) threaten many of its populations across its range. Hybrids of P. perezi and P. ridibundus (P. kl. grafi) can breed successfully with the former parental species after discarding the whole P. perezi genome in the germinal line, thus representing a sexual parasite for P. perezi. However, little is known about the extent of the contact zone of this hybridogenetic complex. Due to the morphological similarity of the three taxa, molecular tools are needed to delineate their respective ranges. Here we characterise a set of 16 microsatellite markers specifically developed for P. perezi. These markers showed moderate to high polymorphism (2–17 alleles/locus) in two populations from central Spain (n=20 and n=23), allowing individual identification of frogs. Seven of these markers cross-amplified in individuals of P. ridibundus from southern France (3–8 alleles/locus). These markers were used to genotype samples along a transect from southern France to eastern Spain, encompassing both pure and hybrid individuals. Sample assignment to each taxon was based on the new microsatellite loci and compared with nuclear and mitochondrial sequence data. Our results show that these markers are useful to distinguish P. ridibundus, P. pereziand the hybrid form P. kl. grafi from each other, even when sample sizes are low. The newly characterised markers will also be useful in demographic and phylogeographic studies in P. perezi and are thus a valuable tool for evolutionary and conservation oriented research, This research was funded by grants CGL2008-04271-C02- 01/BOS, and CGL2011-28300 (Ministerio de Ciencia e Innovación- MICINN), Ministerio de Economía y Competitividad- MEC, Spain, and FEDER). G. Sánchez-Montes is funded by a predoctoral grant provided by the Asociación de Amigos de la Universidad de Navarra. E. Recuero is supported by a DGAPA-UNAM postdoctoral fellowship. J. Gutiérrez-Rodríguez is supported by the Consejo Superior de Investigaciones Científicas of Spain (CSIC) and the European Social Fund (ESF) (JAE-pre PhD fellowship). IMS was funded by the project ‘Biodiversity, Ecology and Global Change’, co-financed by North Portugal Regional Operational Programme 2007/2013 (ON.2–O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF) and is currently supported by funding from the Spanish Severo Ochoa Program (SEV- 2012-0262).

The response of phenolic composition of skins from Vitis vinifera L. 'Tempranillo' and 'Graciano' grapes to water-deficit irrigation during berry growth and ripening was evaluated. The study was carried out using container-grown grapevines grown under controlled conditions in a greenhouse. Two irrigation treatments were imposed: control (well-watered) and sustained deficit irrigation (SDI). Twenty-eight phenolic compounds, including anthocyanins, flavonols and monomeric flavan-3-ols (catechins) as well as phenolic acids derivatives have been identified in the extracts prepared from the berry skins at physiological maturity. For both varieties, water deficit reduced leaf area and leaf area to crop mass ratio, and decreased berry size. However, there were no changes in juice total soluble solids, pH or total polyphenolic content. Water deficit resulted in decreased must titratable acidity in 'Graciano' berries. In 'Tempranillo', water limitation reduced total anthocyanins and flavonols, and increased hydroxycinnamic acids. In 'Graciano', water deficit resulted in increased flavonols and reduced catechins. Altogether, we concluded that under water-deficit irrigation, 'Graciano' grapes presented a differential composition of phenolic compounds that could result in improved fruit quality., This project was supported by Fundación Universitaria de Navarra (2011) and the Ministerio de Ciencia e Innovación (MCINN BFU2011-26989) of Spain. M. NICULCEA was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

Purpose: The objective of this work was to evaluate the effect in the immune response produced by CpG oligodeoxynucleotides (ODN) co-encapsulated with the antigen ovalbumin (OVA) within poly(lactic-co-glycolic) acid (PLGA) 502 and 752 microparticles (MP). Methods: MP were prepared by blending 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) with PLGA and Total Recirculation One Machine System (TROMS) technology and contained OVA along with CpG sequences associated to DOTAP. After confirming the integrity of both encapsulated molecules, BALB/c mice were immunized with the resulting MP and OVA-specific antibodies and cytokine production were assessed in order to determine the immunological profile induced in mice. Results: One μm near non-charged MP co-encapsulated very efficiently both OVA and CpG ODN. The release of both OVA and CpG was slow and incomplete irrespective of polymer. The results of the immune response induced in BALB/c mice indicated that, depending on the PLGA polymer used, co-encapsulation did not improve the immunogenicity of the antigen, compared either with the simply co-administration of both antigen and CpG, or with the microencapsulated antigen. Thus, mice immunized with OVA associated to PLGA 756 displayed an IgG2a characterized response which was biased to an IgG1 profile in case of CpG co-encapsulation. On the contrary, the co-encapsulation of CpG with OVA into PLGA 502 significantly improved the isotype shifting in comparison with the one showed by mice immunized with OVA loaded PLGA 502. Conclusion: This study underlines the importance of MP characteristics to fully exploit simultaneous antigen and CpG ODN particulate delivery as effective vaccine construct., BSR had a grant from Asociación de Amigos de la Universidad de Navarra and the work was financially supported by “Ministerio de Educación y Ciencia” (SAF 2004-07150) and Caja de Ahorros de Navarra (Programa Tú eliges: Tú decides).