Análisis de los factores responsables de la evolución de Angiospermas durante el Cuaternario : un estudio macro- y microevolutivo en "Linaria Sect. Supinae"

Programa de Doctorado en Estudios MedioambientalesEn la presente memoria de tesis, se descubren patrones evolutivos de un grupo de plantas con flor formado por la sect. Supinae del género Linaria (Piantaginaceae). Se presentan cinco manuscritos cientlficos en los que se han utilizado distintos niveles taxonómicos (género, sección, subsección,especie y población) como hipótesis evolutivas. Cada manuscrito tiene objetivos específicos relevantes a distintas escalas espaciales y temporales. Se presta especial atención a las consecuencias evolutivas derivadas de las distintas estrategias reproductivas de las especies. Concretamente tras evaluar la monofilia de la sect. Supinae,se discuten los patrones evolutivos relacionados con la hibridación entre especies, la autogamia y la alogamia;además se analiza el papel de los polinizadores en la evolución de algunas especies alógamas. La introducción comienza describiendo herramientas fundamentales utilizadas en la presente memoria de tesis. Posteriormente se resumen el rol que pueden tener las diversas estrategias reproductivas en la evolución de las plantas con flor. Tras la presentación de cinco manuscritos,los resultados principales de esta investigación se discuten en el contexto de la evolución de plantas con flor durante el Cuaternario.Universidad Pablo de Olavide. Centro de Estudios de Postgrad

Evidence of gastrointestinal sensing and gut-brain communication in rainbow trout (Oncorhynchus mykiss) in response to the aqueous extract of fishmeal and its free amino acid fraction

Using rainbow trout (Oncorhynchus mykiss) as a model, we aimed to obtain information about the gastrointestinal tract (GIT) amino acid sensing capacity and hormone production along regions of the GIT, in response to proline (Pro), to a solution of free amino acids (FAA) mimicking the composition of a fishmeal (FM) aqueous extract (FM-FAA), or to the whole FM aqueous extract (FM-AQE). In addition, we evaluated central responses (in hypothalamus) in mechanisms regulating food intake, 2 h following intragastric administration of these treatments. The presence of Pro in the GIT elicited changes in amino acid sensing systems and in the production of GIT hormones, especially in the more proximal regions in parallel with an anorectic response in hypothalamus. The intragastric administration of FM-AQE induced increased production of the anorectic hormones peptide tyrosine-tyrosine (PYY) and cholecystokinin (CCK) that occurred 20 min post-treatment in the proximal and middle intestine of this treatment. These changes occurred in parallel with an anorectic response in the hypothalamus 2 h post-treatment. The treatment with FM-FAA elicited a comparable anorectic response in the hypothalamus at 2 h post-treatment, which was associated however with a more complex response in the GIT. This included a comparable increased production of the anorectic hormones PYY and CCK in the proximal and middle intestine, but also a decreased production of the orexigenic hormone ghrelin (GHRL) in the stomach, 20 min after FM-FAA administration. These effects were also accompanied by some changes in parameters related to amino acid sensing systems mediated by receptors, which were not observed in the FM-AQE treatment. Overall, results indicate that all treatments elicited a response in elements of gut sensing mechanisms and gut-brain axis, despite important differences in the specific genes (likely having different substrate specificities), GIT areas and times in which responses were observedXunta de Galicia | Ref. ED431B 2022/01Ministerio de Universidades | Ref. FPU19/00122Agencia Estatal de Investigación | Ref. IJC2019-039166-IFinanciado para publicación en acceso aberto: Universidade de Vigo/CISU

Integrated Membrane Process for the Treatment and Reuse of Residual Table Olive Fermentation Brine and Anaerobically Digested Sludge Centrate

[EN] Management of wastewater is a major challenge nowadays, due to increasing water demand, growing population and more stringent regulations on water quality. Wastewaters from food conservation are especially difficult to treat, since they have high salinity and high organic matter concentration. The aim of this work is the treatment of the effluent from a table olive fermentation process (FTOP) with the aim of reusing it once the organic matter is separated. The process proposed in this work consists of the following membrane-based technologies: Ultrafiltration (UF) (UP005, Microdyn Nadir), Forward Osmosis (FO) (Osmen2521, Hydration Technology Innovation) and Nanofiltration (NF) (NF245, Dow). The FO process was implemented to reduce the salinity entering the NF process, using the FTOP as draw solution and, at the same time, to concentrate the centrate produced in the sludge treatment of a municipal wastewater treatment plant with the aim of obtaining a stream enriched in nutrients. The UF step achieved the elimination of 50% of the chemical oxygen demand of the FTOP. The UF permeate was pumped to the FO system reducing the volume of the anaerobically digested sludge centrate (ADSC) by a factor of 3 in 6.5 h. Finally, the ultrafiltrated FTOP diluted by FO was subjected to NF. The transmembrane pressure needed in the NF stage was 40% lower than that required if the ultrafiltration permeate was directly nanofiltered. By means of the integrated process, the concentration of organic matter and phenolic compounds in the FTOP decreased by 97%. Therefore, the proposed process was able to obtain a treated brine that could be reused in other processes and simultaneously to concentrate a stream, such as the ADSC.This research was funded by CDTI (Centre for Industrial and Technological Development) depending on the Spanish Ministry of Science and Innovation through the INNPRONTA program, grant number IPT-20111020.Carbonell Alcaina, C.; Soler-Cabezas, JL.; Bes-Piá, M.; Vincent Vela, MC.; Mendoza Roca, JA.; Pastor-Alcañiz, L.; Alvarez Blanco, S. (2020). Integrated Membrane Process for the Treatment and Reuse of Residual Table Olive Fermentation Brine and Anaerobically Digested Sludge Centrate. Membranes. 10(10):1-14. https://doi.org/10.3390/membranes10100253S1141010Abou-Elela, S. I., Kamel, M. M., & Fawzy, M. E. (2010). Biological treatment of saline wastewater using a salt-tolerant microorganism. Desalination, 250(1), 1-5. doi:10.1016/j.desal.2009.03.022Ferrer-Polonio, E., Iborra-Clar, A., Mendoza-Roca, J. A., & Pastor-Alcañiz, L. (2014). Fermentation brines from Spanish style green table olives processing: treatment alternatives before recycling or recovery operations. Journal of Chemical Technology & Biotechnology, 91(1), 131-137. doi:10.1002/jctb.4550Reid, E., Liu, X., & Judd, S. J. (2006). Effect of high salinity on activated sludge characteristics and membrane permeability in an immersed membrane bioreactor. Journal of Membrane Science, 283(1-2), 164-171. doi:10.1016/j.memsci.2006.06.021Ferrer-Polonio, E., Mendoza-Roca, J. A., Iborra-Clar, A., Alonso-Molina, J. ., & Pastor-Alcañiz, L. (2015). Comparison of two strategies for the start-up of a biological reactor for the treatment of hypersaline effluents from a table olive packaging industry. Chemical Engineering Journal, 273, 595-602. doi:10.1016/j.cej.2015.03.062Li, H., Meng, F., Duan, W., Lin, Y., & Zheng, Y. (2019). Biodegradation of phenol in saline or hypersaline environments by bacteria: A review. 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A., & Pastor-Alcañiz, L. (2018). Ultrafiltration of residual fermentation brines from the production of table olives at different operating conditions. Journal of Cleaner Production, 189, 662-672. doi:10.1016/j.jclepro.2018.04.127Kiai, H., García-Payo, M. C., Hafidi, A., & Khayet, M. (2014). Application of membrane distillation technology in the treatment of table olive wastewaters for phenolic compounds concentration and high quality water production. Chemical Engineering and Processing: Process Intensification, 86, 153-161. doi:10.1016/j.cep.2014.09.007Vu, M. T., Ansari, A. J., Hai, F. I., & Nghiem, L. D. (2018). Performance of a seawater-driven forward osmosis process for pre-concentrating digested sludge centrate: organic enrichment and membrane fouling. Environmental Science: Water Research & Technology, 4(7), 1047-1056. doi:10.1039/c8ew00132dSoler-Cabezas, J. L., Mendoza-Roca, J. A., Vincent-Vela, M. C., Luján-Facundo, M. J., & Pastor-Alcañiz, L. (2018). Simultaneous concentration of nutrients from anaerobically digested sludge centrate and pre-treatment of industrial effluents by forward osmosis. Separation and Purification Technology, 193, 289-296. doi:10.1016/j.seppur.2017.10.058Duan, J., Litwiller, E., Choi, S.-H., & Pinnau, I. (2014). Evaluation of sodium lignin sulfonate as draw solute in forward osmosis for desert restoration. Journal of Membrane Science, 453, 463-470. doi:10.1016/j.memsci.2013.11.029Hu, B., Jiang, M., Zhao, S., Ji, X., Shu, Q., Tian, B., … Zhang, L. (2019). Biogas slurry as draw solution of forward osmosis process to extract clean water from micro-polluted water for hydroponic cultivation. Journal of Membrane Science, 576, 88-95. doi:10.1016/j.memsci.2019.01.029Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. 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Current status and future challenges of table olive processing wastewater valorization. Biochemical Engineering Journal, 112, 103-113. doi:10.1016/j.bej.2016.04.008Carbonell-Alcaina, C., Corbatón-Báguena, M.-J., Álvarez-Blanco, S., Bes-Piá, M. A., Mendoza-Roca, J. A., & Pastor-Alcañiz, L. (2016). Determination of fouling mechanisms in polymeric ultrafiltration membranes using residual brines from table olive storage wastewaters as feed. Journal of Food Engineering, 187, 14-23. doi:10.1016/j.jfoodeng.2016.04.016Sayadi, S., Allouche, N., Jaoua, M., & Aloui, F. (2000). Detrimental effects of high molecular-mass polyphenols on olive mill wastewater biotreatment. Process Biochemistry, 35(7), 725-735. doi:10.1016/s0032-9592(99)00134-xLi, Y., Xu, Z., Xie, M., Zhang, B., Li, G., & Luo, W. (2020). Resource recovery from digested manure centrate: Comparison between conventional and aquaporin thin-film composite forward osmosis membranes. Journal of Membrane Science, 593, 117436. doi:10.1016/j.memsci.2019.117436Bargeman, G., Vollenbroek, J. M., Straatsma, J., Schroën, C. G. P. H., & Boom, R. M. (2005). Nanofiltration of multi-component feeds. Interactions between neutral and charged components and their effect on retention. Journal of Membrane Science, 247(1-2), 11-20. doi:10.1016/j.memsci.2004.05.022Koyuncu, I., Topacik, D., & Wiesner, M. R. (2004). Factors influencing flux decline during nanofiltration of solutions containing dyes and salts. Water Research, 38(2), 432-440. doi:10.1016/j.watres.2003.10.001Virga, E., Spruijt, E., de Vos, W. M., & Biesheuvel, P. M. (2018). Wettability of Amphoteric Surfaces: The Effect of pH and Ionic Strength on Surface Ionization and Wetting. Langmuir, 34(50), 15174-15180. doi:10.1021/acs.langmuir.8b02875Van der Bruggen, B., Cornelis, G., Vandecasteele, C., & Devreese, I. (2005). 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The Colonization History of Juniperus brevifolia (Cupressaceae) in the Azores Islands

Background A central aim of island biogeography is to understand the colonization history of insular species using current distributions, fossil records and genetic diversity. Here, we analyze five plastid DNA regions of the endangered Juniperus brevifolia, which is endemic to the Azores archipelago. Methodology/Principal Findings The phylogeny of the section Juniperus and the phylogeographic analyses of J. brevifolia based on the coalescence theory of allele (plastid) diversity suggest that: (1) a single introduction event likely occurred from Europe; (2) genetic diversification and inter-island dispersal postdated the emergence of the oldest island (Santa Maria, 8.12 Ma); (3) the genetic differentiation found in populations on the islands with higher age and smaller distance to the continent is significantly higher than that on the younger, more remote ones; (4) the high number of haplotypes observed (16), and the widespread distribution of the most frequent and ancestral ones across the archipelago, are indicating early diversification, demographic expansion, and recurrent dispersal. In contrast, restriction of six of the seven derived haplotypes to single islands is construed as reflecting significant isolation time prior to colonization. Conclusions/Significance Our phylogeographic reconstruction points to the sequence of island emergence as the key factor to explain the distribution of plastid DNA variation. The reproductive traits of this juniper species (anemophily, ornithochory, multi-seeded cones), together with its broad ecological range, appear to be largely responsible for recurrent inter-island colonization of ancestral haplotypes. In contrast, certain delay in colonization of new haplotypes may reflect intraspecific habitat competition on islands where this juniper was already present.B. Rumeu and J.L. Blanco-Pastor were funded by a Spanish National Research Council grant µ(CSIC: JAE-PRE; http://www.csic.es/web/guest/programa-ja​e). This contribution is framed within the projects CGL2010-18759 and PI2007/053 and financed by the Spanish Ministry of Science and Innovation (http://www.micinn.es), and the Canary Island Government (http://www.gobcan.es), respectively, both of which were partially funded by the European Union. The Organismo Autónomo de Parques Nacionales (http://www.mma.es/secciones/el_ministeri​o/organismos/oapn/) also financed part of this work (project 051/2010). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewe

Annual and perennial Medicago show signatures of parallel adaptation to climate and soil in highly conserved genes

Human induced environmental change may require rapid adaptation of plant populations and crops, but the genomic basis of environmental adaptation remain poorly understood. We analysed polymorphic loci from the perennial crop Medicago sativa (alfalfa or lucerne) and the annual legume model species M. truncatula to search for a common set of candidate genes that might contribute to adaptation to abiotic stress in both annual and perennial Medicago species. We identified a set of candidate genes of adaptation associated with environmental gradients along the distribution of the two Medicago species. Candidate genes for each species were detected in homologous genomic linkage blocks using genome-environment (GEA) and genome-phenotype association analyses. Hundreds of GEA candidate genes were species-specific, of these, 13.4% (M. sativa) and 24% (M. truncatula) were also significantly associated with phenotypic traits. A set of 168 GEA candidates were shared by both species, which was 25.4% more than expected by chance. When combined, they explained a high proportion of variance for certain phenotypic traits associated with adaptation. Genes with highly conserved functions dominated among the shared candidates and were enriched in gene ontology terms that have shown to play a central role in drought avoidance and tolerance mechanisms by means of cellular shape modifications and other functions associated with cell homeostasis. Our results point to the existence of a molecular basis of adaptation to abiotic stress in Medicago determined by highly conserved genes and gene functions. We discuss these results in light of the recently proposed omnigenic model of complex traits

Pleistocene climate changes, and not agricultural spread, accounts for range expansion and admixture in the dominant grassland species <i>Lolium perenne</i> L.

International audienceAim: Grasslands have been pivotal in the development of herbivore breeding since the Neolithic and still represent the most widespread agricultural land use across Europe. However, it remains unclear whether the current large‐scale genetic variation of plant species found in natural grasslands of Europe is the result of human activities or natural processes. Location: Europe. Taxon: Lolium perenne L. (perennial ryegrass). Methods: We reconstructed the phylogeographic history of L. perenne, a dominant grassland species, using 481 natural populations, including 11 populations of closely related taxa. We combined Genotyping‐by‐Sequencing (GBS) and pool‐Sequencing (pool‐Seq) to obtain high‐quality allele frequency calls of ~500 k SNP loci. We performed genetic structure analyses and demographic reconstructions based on the site frequency spectrum (SFS). We additionally used the same genotyping protocol to assess the genomic diversity of a set of 32 cultivars representative of the L. perenne cultivars widely used for forage purposes. Results: Expansion across Europe took place during the Würm glaciation (12–110 kya), a cooling period that decreased the dominance of trees in favour of grasses. Splits and admixtures in L. perenne fit historical climate changes in the Mediterranean basin. The development of agriculture in Europe (7–3.5 kya), that caused an increase in the abundance of grasslands, did not have an effect on the demographic patterns of L. perenne. We found that most modern cultivars are closely related to natural diversity from north-western Europe. Thus, modern cultivars do not represent the wide genetic variation found in natural populations. Main conclusions: Demographic events in L. perenne can be explained by the changing climatic conditions during the Pleistocene. Natural populations maintain a wide genomic variability at continental scale that has been minimally exploited by recent breeding activities. This variability constitutes valuable standing genetic variation for future adaptation of grasslands to climate change, safeguarding the agricultural services they provide

Development and Characterization of a Tacrolimus/Hydroxypropyl-β-Cyclodextrin Eye Drop

Uveitis is a vision inflammatory disorder with a high prevalence in developing countries. Currently, marketed treatments remain limited and reformulation is usually performed to obtain a tacrolimus eye drop as a therapeutic alternative in corticosteroid-refractory eye disease. The aim of this work was to develop a mucoadhesive, non-toxic and stable topical ophthalmic formulation that can be safely prepared in hospital pharmacy departments. Four different ophthalmic formulations were prepared based on the tacrolimus/hydroxypropyl-β-cyclodextrin (HPβCD) inclusion complexes’ formation. Phase solubility diagrams, Nuclear Magnetic Resonance (NMR) and molecular modeling studies showed the formation of 1:1 and 1:2 tacrolimus/HPβCD inclusion complexes, being possible to obtain a 0.02% (w/v) tacrolimus concentration by using 40% (w/v) HPβCD aqueous solutions. Formulations also showed good ophthalmic properties in terms of pH, osmolality and safety. Stability studies proved these formulations to be stable for at least 3 months in refrigeration. Ex vivo bioadhesion and in vivo ocular permanence showed good mucoadhesive properties with higher ocular permanence compared to the reference pharmacy compounding used in clinical settings (t1/2 of 86.2 min for the eyedrop elaborated with 40% (w/v) HPβCD and Liquifilm® versus 46.3 min for the reference formulation). Thus, these novel eye drops present high potential as a safe alternative for uveitis treatment, as well as a versatile composition to include new drugs intended for topical ophthalmic administrationThis research was partially supported by the Spanish Ministry of Science, Innovation and Universities (RTI2018-099597-B-100), the ISCIII (PI17/00940, RETICS Oftared, RD16/0008/0003 and RD12/0034/0017) and by Xunta de Galicia, grant numbers GPC2013/015 and GRC2017/015S

High-Throughput Genome-Wide Genotyping To Optimize the Use of Natural Genetic Resources in the Grassland Species Perennial Ryegrass (Lolium perenne L.)

The natural genetic diversity of agricultural species is an essential genetic resource for breeding programs aiming to improve their ecosystem and production services. A large natural ecotype diversity is usually available for most grassland species. This could be used to recombine natural climatic adaptations and agronomic value to create improved populations of grassland species adapted to future regional climates. However describing natural genetic resources can be long and costly. Molecular markers may provide useful information to help this task. This opportunity was investigated for Lolium perenne L., using a set of 385 accessions from the natural diversity of this species collected right across Europe and provided by genebanks of several countries. For each of these populations, genotyping provided the allele frequencies of 189,781 SNP markers. GWAS were implemented for over 30 agronomic and/or putatively adaptive traits recorded in three climatically contrasted locations (France, Belgium, Germany). Significant associations were detected for hundreds of markers despite a strong confounding effect of the genetic background; most of them pertained to phenology traits. It is likely that genetic variability in these traits has had an important contribution to environmental adaptation and ecotype differentiation. Genomic prediction models calibrated using natural diversity were found to be highly effective to describe natural populations for almost all traits as well as commercial synthetic populations for some important traits such as disease resistance, spring growth or phenological traits. These results will certainly be valuable information to help the use of natural genetic resources of other species

Canonical correlations reveal adaptive loci and phenotypic responses to climate in perennial ryegrass

Germplasm from perennial ryegrass (Lolium perenne L.) natural populations is useful for breeding because of its adaptation to a wide range of climates. Climate‐adaptive genes can be detected from associations between genotype, phenotype and climate but an integrated framework for the analysis of these three sources of information is lacking. We used two approaches to identify adaptive loci in perennial ryegrass and their effect on phenotypic traits. First, we combined Genome‐Environment Association (GEA) and GWAS analyses. Then, we implemented a new test based on a Canonical Correlation Analysis (CANCOR) to detect adaptive loci. Furthermore, we improved the previous perennial ryegrass gene set by de novo gene prediction and functional annotation of 39,967 genes. GEA‐GWAS revealed eight outlier loci associated with both environmental variables and phenotypic traits. CANCOR retrieved 633 outlier loci associated with two climatic gradients, characterized by cold‐dry winter versus mild‐wet winter and long rainy season versus long summer, and pointed out traits putatively conferring adaptation at the extremes of these gradients. Our CANCOR test also revealed the presence of both polygenic and oligogenic climatic adaptations. Our gene annotation revealed that 374 of the CANCOR outlier loci were positioned within or close to a gene. Co‐association networks of outlier loci revealed a potential utility of CANCOR for investigating the interaction of genes involved in polygenic adaptations. The CANCOR test provides an integrated framework to analyse adaptive genomic diversity and phenotypic responses to environmental selection pressures that could be used to facilitate the adaptation of plant species to climate change

A Guide to Carrying Out a Phylogenomic Target Sequence Capture Project

High-throughput DNA sequencing techniques enable time- and cost-effective sequencing of large portions of the genome. Instead of sequencing and annotating whole genomes, many phylogenetic studies focus sequencing effort on large sets of pre-selected loci, which further reduces costs and bioinformatic challenges while increasing coverage. One common approach that enriches loci before sequencing is often referred to as target sequence capture. This technique has been shown to be applicable to phylogenetic studies of greatly varying evolutionary depth. Moreover, it has proven to produce powerful, large multi-locus DNA sequence datasets suitable for phylogenetic analyses. However, target capture requires careful considerations, which may greatly affect the success of experiments. Here we provide a simple flowchart for designing phylogenomic target capture experiments. We discuss necessary decisions from the identification of target loci to the final bioinformatic processing of sequence data. We outline challenges and solutions related to the taxonomic scope, sample quality, and available genomic resources of target capture projects. We hope this review will serve as a useful roadmap for designing and carrying out successful phylogenetic target capture studies. © Copyright © 2020 Andermann, Torres Jiménez, Matos-Maraví, Batista, Blanco-Pastor, Gustafsson, Kistler, Liberal, Oxelman, Bacon and Antonelli