In-depth analysis of the Quercus suber metabolome under drought stress and recovery reveals potential key metabolic players

Cork oak (Quercus suber L.) is a species of ecological, social and economic importance in the Mediterranean region. Given its xerophytic adaptability, the study of cork oak's response to drought stress conditions may provide important data in the global scenario of climate change. The mechanisms behind cork oak's adaptation to drought conditions can inform the design and development of tools to better manage this species under the changing climate patterns. Metabolomics is one of the most promising omics layers to capture a snapshot of a particular physiological state and to identify putative biomarkers of stress tolerance. Drastic changes were observed in the leaf metabolome of Q. suber between the different experimental conditions, namely at the beginning of the drought stress treatment, after one month under drought and post rehydration. All experimental treatments were analyzed through sPLS to inspect for global changes and stress and rehydration responses were analyzed independently for specific alterations. This allowed a more in-depth study and a search for biomarkers specific to a given hydric treatment. The metabolome analyses showed changes in both primary and secondary metabolism, but highlighted the role of secondary metabolism. In addition, a compound-specific response was observed in stress and rehydration. Key compounds such as L-phenylalanine and epigallocatechin 3-gallate were identified in relation to early drought response, terpenoid leonuridine and the flavonoid glycoside (-)-epicatechin-3'-O-glucuronide in long-term drought response, and flavone isoscoparine was identified in relation to the recovery process. The results here obtained provide novel insights into the biology of cork oak, highlighting pathways and metabolites potentially involved in the response of this species during drought and recovery that may be essential for its adaptation to long periods of drought. It is expected that this knowledge can encourage further functional studies in order to validate potential biomarkers of drought and recovery that maybe used to support decision-making in cork oak breeding programs.publishe

Conserved epigenetic mechanisms could play a key role in regulation of photosynthesis and development-related genes during needle development of Pinus radiata

Needle maturation is a complex process that involves cell growth, differentiation and tissue remodelling towards the acquisition of full physiological competence. Leaf induction mechanisms are well known; however, those underlying the acquisition of physiological competence are still poorly understood, especially in conifers. We studied the specific epigenetic regulation of genes defining organ function (PrRBCS and PrRBCA) and competence and stress response (PrCSDP2 and PrSHMT4) during three stages of needle development and one de-differentiated control. Gene-specific changes in DNA methylation and histone were analysed by bisulfite sequencing and chromatin immunoprecipitation (ChIP). The expression of PrRBCA and PrRBCS increased during needle maturation and was associated with the progressive loss of H3K9me3, H3K27me3 and the increase in AcH4. The maturation-related silencing of PrSHMT4 was correlated with increased H3K9me3 levels, and the repression of PrCSDP2, to the interplay between AcH4, H3K27me3, H3K9me3 and specific DNA methylation. The employ of HAT and HDAC inhibitors led to a further determination of the role of histone acetylation in the regulation of our target genes. The integration of these results with high-throughput analyses in Arabidopsis thaliana and Populus trichocarpa suggests that the specific epigenetic mechanisms that regulate photosynthetic genes are conserved between the analysed species

Molecular Research on Stress Responses in Quercus spp.: From Classical Biochemistry to Systems Biology through Omics Analysis

The genus Quercus (oak), family Fagaceae, comprises around 500 species, being one of the most important and dominant woody angiosperms in the Northern Hemisphere. Nowadays, it is threatened by environmental cues, which are either of biotic or abiotic origin. This causes tree decline, dieback, and deforestation, which can worsen in a climate change scenario. In the 21st century, biotechnology should take a pivotal role in facing this problem and proposing sustainable management and conservation strategies for forests. As a non-domesticated, long-lived species, the only plausible approach for tree breeding is exploiting the natural diversity present in this species and the selection of elite, more resilient genotypes, based on molecular markers. In this direction, it is important to investigate the molecular mechanisms of the tolerance or resistance to stresses, and the identification of genes, gene products, and metabolites related to this phenotype. This research is being performed by using classical biochemistry or the most recent omics (genomics, epigenomics, transcriptomics, proteomics, and metabolomics) approaches, which should be integrated with other physiological and morphological techniques in the Systems Biology direction. This review is focused on the current state-of-the-art of such approaches for describing and integrating the latest knowledge on biotic and abiotic stress responses in Quercus spp., with special reference to Quercus ilex, the system on which the authors have been working for the last 15 years. While biotic stress factors mainly include fungi and insects such as Phytophthora cinnamomi, Cerambyx welensii, and Operophtera brumata, abiotic stress factors include salinity, drought, waterlogging, soil pollutants, cold, heat, carbon dioxide, ozone, and ultraviolet radiation. The review is structured following the Central Dogma of Molecular Biology and the omic cascade, from DNA (genomics, epigenomics, and DNA-based markers) to metabolites (metabolomics), through mRNA (transcriptomics) and proteins (proteomics). An integrated view of the different approaches, challenges, and future directions is critically discussed

Metabolome Integrated Analysis of High-Temperature Response in Pinus radiata

The integrative omics approach is crucial to identify the molecular mechanisms underlying high-temperature response in non-model species. Based on future scenarios of heat increase, Pinus radiata plants were exposed to a temperature of 40 degrees C for a period of 5 days, including recovered plants (30 days after last exposure to 40 degrees C) in the analysis. The analysis of the metabolome using complementary mass spectrometry techniques (GC-MS and LC-Orbitrap-MS) allowed the reliable quantification of 2,287 metabolites. The analysis of identified metabolites and highlighter metabolic pathways across heat time exposure reveal the dynamism of the metabolome in relation to high-temperature response in P. radiata, identifying the existence of a turning point (on day 3) at which P. radiata plants changed from an initial stress response program (shorter-term response) to an acclimation one (longer-term response). Furthermore, the integration of metabolome and physiological measurements, which cover from the photosynthetic state to hormonal profile, suggests a complex metabolic pathway interaction network related to heat-stress response. Cytokinins (CKs), fatty acid metabolism and flavonoid and terpenoid biosynthesis were revealed as the most important pathways involved in heat-stress response in P. radiata, with zeatin riboside (ZR) and isopentenyl adenosine (iPA) as the key hormones coordinating these multiple and complex interactions. On the other hand, the integrative approach allowed elucidation of crucial metabolic mechanisms involved in heat response in P. radiata, as well as the identification of thermotolerance metabolic biomarkers (L-phenylalanine, hexadecanoic acid, and dihydromyricetin), crucial metabolites which can reschedule the metabolic strategy to adapt to high temperature

Kaolin and salicylic acid alleviate summer stress in rainfed olive orchards by modulation of distinct physiological and biochemical responses

In a changing world, the search for new agronomic practices that help crops to maintain and/or increase yields and quality is a continuous challenge. We aim to evaluate kaolin (KL) and salicylic acid (SA) effectiveness as summer stress alleviating agents through physiological, biochemical and immunohistochemical analysis. Olive trees (Olea europaea L. cv. Cobrançosa) grown under rainfed conditions were sprayed with 5% KL and 100 μM SA, at the beginning of summer, during two consecutive years. KL enhanced relative water content (RWC), stomatal conductance (gs) net photosynthesis (A) and leaf indole-3-acetic acid (IAA) signal, and decreased leaf sclerophylly, secondary metabolites and non-structural carbohydrates accumulation and abscisic acid (ABA).The trees treated with SA showed changes on IAA and ABA dynamics, and an enhancement in RWC, gs, A, soluble proteins, and leaf P and Mg concentrations during the summer. Notably, KL and SA also allowed a faster restauration of the physiological functions during stress relief. In sum, KL and SA foliar sprays alleviated the negative effects induced by summer stress in olive trees performance, by modulation of distinct physiological and biochemical responses.info:eu-repo/semantics/publishedVersio

Kaolin spray induces changes in ABA and IAA immunodistribution in olive leaves.

The climate change scenarios predicted the accentuation of drought and high temperature events during the summer season in the Mediterranean region, coming up harmful consequences to important crops of this region, such as olive tree (Olea europaea L.). Stress conditions often stimulate changes in plants production, distribution or signal transduction of phytohormones as a response, then modifying their physiology and biochemistry. Kaolin (KL) is a reflecting clay that applied on leaves surface reduce the common damages promoted by heat load and high irradiance levels, being important to test its effect on olive tree hormonal dynamics and physiological parameters. Olive trees of a rainfed orchard in Northeast Portugal, were sprayed with kaolin 5% (KL) and water (C). The immunodistribution of two hormones, abscisic acid (ABA) and indoleacetic acid (IAA), and some physiological and growth responses were accessed. The ABA signal was substantially more pronounced than the IAA signal in all the analyzed leaves, concomitant with the stressful conditions of which these plants were subjected. In general, ABA signal showed a uniform distribution throughout the leaf in both treatments. However, its intensity was higher in C than in KL-sprayed leaves, reflecting the better water status and higher stomatal conductance of the last plants. While in C leaves was observed a uniform distribution of IAA signal trough the leaf limb and an almost absence of signal in the main vascular tissues, in KL-sprayed leaves was observed a higher signal intensity in the main vascular tissues and upper palisade parenchyma. These data suggest its transport and an active growth in KL plants, concomitant with the higher increase of canopy volume. Thus, immunodetection appears to be an efficient tool to understand the translocation of IAA and ABA in plants treated with abiotic stress alleviating products and clarify their role in regulating the physiological responses.info:eu-repo/semantics/publishedVersio

Kaolin and salicylic acid alleviate summer stress effects on rainfed olive orchards through distinct physiological and biochemical processes

In a changing world, the search for new agronomic practices that help crops to maintain and/or increase yields and quality is a continuous challenge. Olive trees cultivated under rainfed conditions were sprayed with 5% kaolin (KL) and 100 μM salicylic acid (SA) during two consecutive years in the beginning of the summer season. Exogenous KL enhanced relative water content (RWC), stomatal conductance (gs) net photosynthesis (A) and IAA immunodetection, and decreased leaf sclerophylly, secondary metabolites and non-structural carbohydrates accumulation, ABA signal and DNA methylation, contributing to higher growth and yield. The plants treated with SA showed an enhancement in RWC, gs, A, soluble proteins, IAA, ABA and DNA methylation immunodetection and leaf P and Mg concentrations during the summer, leading to higher yield. Thus, KL and SA alleviated some of the negative effects induced by summer stress in olive tree performance, allowing a faster restauration of the physiological functions during the stress relief and leading to higher yieldsThis work was funded by the INTERACT project – “Integrative Research in Environment, Agro-Chains and Technology”, no. NORTE-01-0145-FEDER-000017, in its lines of research entitled ISAC, co-financed by the European Regional Development Fund (ERDF) through NORTE 2020 (North Regional Operational Program 2014/2020).info:eu-repo/semantics/publishedVersio

31 visiones actuales de la transparencia

Treinta y una aportaciones de autores nacionales y extranjeros sobre la transparencia de las instituciones públicas y de los sujetos obligados por la Ley 19/2013 de 9 de diciembre y el derecho de acceso a la información pública

Laboratorio en abierto: aprendendiendo a copiar ADN.2

El objetivo principal del proyecto es la puesta a punto de recursos educativos en abierto (REA) dirigidos a los alumnos de secundaria. La propuesta pretende desarrollar habilidades, para la resolución de problemas científicos, a través de retos que despierten el interés y la imaginación de los alumnos de secundaria. En esta propuesta la resolución de los problemas planteados estaría basada en la aplicación de una herramienta que ha revolucionado la genética y biología, la reacción en Cadena de la Polimerasa, conocida como PCR