Caracterización molecular y funcional del gen PATHOGEN AND CIRCADIAN CONTROLLED 1 (PCC1) en Arabidopsis thaliana

Las plantas son capaces de modificar los patrones de desarrollo tras percibir ciertos tipos de estrés. En Arabidopsis, se identificó PCC1 como un regulador positivo de la transición floral en respuesta al estrés generado por irradiación con luz UV-C. El análisis de plantas transgénicas pPCC1::GUS muestra que PCC1 se expresa durante los primeros días de desarrollo en estomas y haces vasculares de cotiledones. En hojas verdaderas en formación se detecta tinción GUS en su parte basal, incluyendo los haces vasculares, y se va extendiendo completamente a toda la superficie de hojas completamente formadas. Líneas que expresan construcciones de RNAi para PCC1 (iPCC1) presentan reducidos niveles de FT y, consecuentemente, una floración más tardía. El mecanismo por el cual PCC1 podría regular la transición floral parece estar relacionado con alteraciones en la transmisión de la señal por luz. Concomitantemente, las plantas iPCC1 muestran fenotipos parcialmente escotomorfogénicos en los distintos tipos de luz ensayados de forma independiente de la acumulación y señalización de GAs. El transcriptoma diferencial de plantas iPCC1 versus plantas silvestres muestra una clara implicación de PCC1 en procesos relacionados con defensa. De acuerdo con este hecho, hemos observado que las plantas iPCC1 son más susceptibles a la infección con el oomiceto hemi-biotrofo Phytophtora brassicae y más resistentes al hongo necrotrofo Botrytis cinerea. Además, las líneas iPCC1 presentan una regulación al alza de genes de respuesta a ABA, y una mayor sensibilidad a esta fitohormona para los distintos fenotipos analizados. Finalmente, entre los genes alterados en las líneas iPCC1 se observa una sobrerepresentación de genes implicados en el metabolismo y en el transporte de lípidos. La pérdida de función de PCC1 conlleva una reducción del 70% en los niveles de fosfatidilinositol, y en menor medida de otros tipos de lípidos polares como la fosfatidilserina o la fosfatidilcolina. Además, el análisis de la composición de ácidos grasos de cada tipo de lípidos polares revela un mayor grado de insaturación de sus cadenas laterales, fundamentalmente en la fosfatidilserina y el fosfatidilinositol. PCC1 es una proteína asociada a la membrana plasmática por su extremo carboxiterminal, el cual es responsable además de la formación de homodímeros. Aunque queda por dilucidiar los mecanismos por los cuales PCC1 puede regular procesos tan dispares molecularmente como la respuesta a patógenos y la transición floral, hemos observado que PCC1 interacciona con la subunidad CSN5 del signalosoma (CSN), lo que sugiere que PCC1 podría actuar como un regulador de la función de CSN, y en última instancia, de la degradación de proteínas por ubiquitinación.Mir Moreno, R. (2013). Caracterización molecular y funcional del gen PATHOGEN AND CIRCADIAN CONTROLLED 1 (PCC1) en Arabidopsis thaliana [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/29751TESI

Doubled haploids in eggplant

[EN] This review compiles the most relevant advances made in the production of doubled haploid plants in eggplant, the application of doubled haploid lines in breeding programs, and the future perspectives for the development of alternative technologies for doubled haploid generation in this species. Eggplant is a solanaceous crop cultivated worldwide for its edible fruit. Eggplant breeding programs are mainly aimed to the generation of F1 hybrids by crossing two highly homozygous, pure lines, which are traditionally obtained upon several self crossing generations, which is an expensive and time consuming process. Alternatively, fully homozygous, doubled haploid (DH) individuals can be induced from haploid cells of the germ line in a single generation. Several attempts have been made to develop protocols to produce eggplant DHs principally using anther culture and isolated microspore culture. Eggplant could be considered a moderately recalcitrant species in terms of ability for DH production. Anther culture stands nowadays as the most valuable technology to obtain eggplant DHs. However, the theoretical possibility of having plants regenerated from somatic tissues of the anther walls cannot be ruled out. For this reason, the use of isolated microspores is recommended when possible. This approach still has room for improvement, but it is largely genotype-dependent. In this review, we compile the most relevant advances made in DH production in eggplant, their application to breeding programs, and the future perspectives for the development of other, less genotype-dependent, DH technologies.This research was funded by the Valencian Government, grant number CDEIGENT 2018/023 to RMM and by the Spanish MICINN, grant number PID2020-115763RB-I00 to JMSS. ACS is the recipient of a predoctoral contract from the FPU program of the Spanish Government.Mir Moreno, R.; Calabuig-Serna, A.; Seguí-Simarro, JM. (2021). Doubled haploids in eggplant. Biology. 10(7):1-16. https://doi.org/10.3390/biology10070685S11610

The Highly Embryogenic Brassica napus DH4079 Line Is Recalcitrant to Agrobacterium-Mediated Genetic Transformation

[EN] Brassica napus is a species of high agronomic interest, used as a model to study different processes, including microspore embryogenesis. The DH4079 and DH12075 lines show high and low em-bryogenic response, respectively, which makes them ideal to study the basic mechanisms control-ling embryogenesis induction. Therefore, the availability of protocols for genetic transformation of these two backgrounds would help to generate tools to better understand this process. There are some reports in the literature showing the stable transformation of DH12075. However, no equivalent studies in DH4079 have been reported to date. We explored the ability of DH4079 plants to be genetically transformed. As a reference to compare with, we used the same protocols to transform DH12075. We used three different protocols previously reported as successful for B. napus stable transformation with Agrobacterium tumefaciens and analyzed the response of plants. Whereas DH12075 plants responded to genetic transformation, DH4079 plants were completely recalcitrant, not producing any single regenerant out of the 1784 explants transformed and cul-tured. Additionally, an Agrobacterium rhizogenes transient transformation assay was performed on both lines, and only DH12075, but no DH4079 seedlings, responded to A. rhizogenes infection. Therefore, we propose that the DH4079 line is recalcitrant to Agrobacterium-mediated transfor-mation.This research was funded by MCIN/AEI/10.13039/501100011033, grant number PID2020-115763RB-I00 to JMSS and by Generalitat Valenciana, grant number CDEIGENT 2018/023 to RM. ACS is recipient of a PhD contract (FPU17/00715) from the Spanish Ministerio de Educacion.Calabuig-Serna, A.; Mir Moreno, R.; Porcel, R.; Seguí-Simarro, JM. (2023). The Highly Embryogenic Brassica napus DH4079 Line Is Recalcitrant to Agrobacterium-Mediated Genetic Transformation. Plants. 12(10). https://doi.org/10.3390/plants12102008121

Salt and Water Stress Tolerance in Ipomoea purpurea and Ipomoea tricolor, Two Ornamentals with Invasive Potential

[EN] Invasive plants pose a significant threat to biodiversity, especially under the current unstable climatic conditions. This study aimed to test the salt and drought tolerance of two ornamental species of the genus Ipomoea during germination and vegetative growth. Germination tests were performed in the presence of increasing NaCl concentrations or iso-osmotic PEG concentrations¿to mimic the osmotic stress caused by drought. Both species showed great invasive potential because of their high seed germination percentages and rapid germination under control (distilled water) and salt stress conditions, up to 200 mM NaCl. Germination and early seedling development were more affected in the presence of PEG. Subsequently, water stress (complete withholding of irrigation) and salt stress (watering with 100 mM and 200 mM NaCl) treatments were applied to young plants for three weeks, when all plants were harvested, to determine several morphological and biochemical parameters. Both species were sensitive to water deficit but relatively resistant to salt stress. Their salt stress responses were similar, based mainly on the inhibition of Na+ and the activation of K+ transport from roots to leaves and the uptake and accumulation of Ca2+; however, I. tricolor showed a slightly higher tolerance to salt stress than I. purpurea. Although I. tricolor has only been locally reported as invasive and is generally considered a `low-risk¿ species, our results indicate that it may have an invasive potential even higher than I. purpurea, a recognised invasive weed, spread into areas with moderate salinity, affecting agricultural land or natural habitats of ecological interestD.-M.M. is supported by a pre-doctoral contract from the Polytechnic University of Valencia, Spain and R.M. by a CDEIGENT (2018/2023) grant from Generalitat Valenciana.Mircea, DM.; Li, R.; Blasco Giménez, L.; Vicente, O.; Sestras, AF.; Sestras, RE.; Boscaiu, M.... (2023). Salt and Water Stress Tolerance in Ipomoea purpurea and Ipomoea tricolor, Two Ornamentals with Invasive Potential. Agronomy. 13(9). https://doi.org/10.3390/agronomy1309219813

Physiological and Biochemical Responses to Water Stress and Salinity of the Invasive Moth Plant, Araujia sericifera Brot., during Seed Germination and Vegetative Growth

[EN] Araujia sericifera is an invasive plant with an increasing presence in South East Spain, where it produces damage to native trees and shrubs and citric orchards. As the climatic conditions in the study area are becoming harsher due to the climate change, the stress tolerance of this species has been studied during germination and vegetative growth. Growth parameters, photosynthetic pigments, ion accumulation, and antioxidant mechanisms were analysed in plants that were subjected to water deficit and salt stress. Seed germination was reduced by salinity but 50% of the seeds still germinated at 50 mM NaCl. The ungerminated seeds did not lose their germination capacity as shown in `recovery¿ germination assays in distilled water. Germination was less affected by osmotic stress that was induced by polyethylene glycol (PEG), and germination velocity increased in the recovery treatments after exposure to NaCl or PEG. Plant growth was practically unaffected by 150 mM NaCl but inhibited by higher NaCl concentrations or severe drought stress. Nevertheless, all the plants survived throughout the experiment, even under high salinity (600 mM NaCl). A. sericifera relative stress tolerance relies, at least to some extent, on effective antioxidant mechanisms that are based on flavonoid biosynthesis and the activation of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, and glutathione reductase.Bellache, M.; Moltó, N.; Allal Benfekih, L.; Torres-Pagán, N.; Mir Moreno, R.; Verdeguer Sancho, MM.; Boscaiu, M.... (2022). Physiological and Biochemical Responses to Water Stress and Salinity of the Invasive Moth Plant, Araujia sericifera Brot., during Seed Germination and Vegetative Growth. Agronomy. 11(2):1-20. https://doi.org/10.3390/agronomy1202036112011

Multidrug resistance protein 1 localization in lipid raft domains and prostasomes in prostate cancer cell lines

Background: One of the problems in prostate cancer (CaP) treatment is the appearance of the multidrug resistance phenotype, in which ATP-binding cassette transporters such as multidrug resistance protein 1 (MRP1) play a role. Different localizations of the transporter have been reported, some of them related to the chemoresistant phenotype. Aim: This study aimed to compare the localization of MRP1 in three prostate cell lines (normal, androgen-sensitive, and androgen-independent) in order to understand its possible role in CaP chemoresistance. Methods: MRP1 and caveolae protein markers were detected using confocal microscopy, performing colocalization techniques. Lipid raft isolation made it possible to detect these proteins by Western blot analysis. Caveolae and prostasomes were identified by electron microscopy. Results: We show that MRP1 is found in lipid raft fractions of tumor cells and that the number of caveolae increases with malignancy acquisition. MRP1 is found not only in the plasma membrane associated with lipid rafts but also in cytoplasmic accumulations colocalizing with the prostasome markers Caveolin-1 and CD59, suggesting that in CaP cells, MRP1 is localized in prostasomes. Conclusion: We hypothesize that the presence of MRP1 in prostasomes could serve as a reservoir of MRP1; thus, taking advantage of the release of their content, MRP1 could be translocated to the plasma membrane contributing to the chemoresistant phenotype. The presence of MRP1 in prostasomes could serve as a predictor of malignancy in Ca

Bioactive compounds recovery optimization from vine pruning residues using conventional heating and microwave-assisted extraction methods

Polyphenol compounds from vine pruning residue (VPR) were extracted by conventional heating and microwave-assisted treatments. For each treatment, total phenolic compounds and their antioxidant activity were optimized by experimental design. Maximal extraction of polyphenolic compounds (2.17g/100g VPR) was obtained at 80°C, 120min and 45% of ethanol by conventional heating, and 2.37g/100g of VPR were extracted by microwave-assisted process at 120°C, 5min and 60% of ethanol. Ellagic acid and apigenin were the predominant polyphenolic compounds in the extracts, achieving concentration of 68.65 and 208.23mg/100g VPR, respectively for conventional heating and 185.15 and 118.84mg/100g of VPR for microwave-assisted treatment. The results showed reduction of extraction time and energy consumption for microwave-assisted treatment leading to cost-effective technology for the extraction of polyphenol compounds. Furthermore, the results hereby compiled allow for the tailor-made extraction of specific high-value compounds from a renewable biomass as vine pruning residue.Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unitBioTecNorte operation(NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do NorteInternational Cooperation Program CNPq/CSFinfo:eu-repo/semantics/publishedVersio

Los Batanes: A trap for the Pyrenean wild goat during the Late Pleistocene (Spain)

Los Batanes cave (1025 m.a.s.l.) is a karstic system formed by pressure ducts, near the locality of Biescas in the Upper Gallego Valley (Huesca, Spain). Many fossil remains were retrieved from the cave sediments. The minimum sediment calendar age was determined to be 12.770 ± 60 BP. This date indicates that the deposit was formed before the Late Pleistocene-Early Holocene transition at the Pyrenees. Here we analyze the faunal assemblage of the site that is only conformed by remains of Pyrenean wild goat (Capra pyrenaica pyrenaica), which is a recently extinct subspecies of Iberian wild goat (C. pyrenaica). In the site, 1079 remains of Iberian wild goat have been recovered being one of the most numerous both in minimum number of individuals (MNI) and number of identified specimens (NISP) recovered until now in a natural trap in the Spanish Pyrenees. The population of Los Batanes shows a wide range of ages between juvenile and senile individuals and the MNI estimated from the number of right metatarsus is nine. The taphonomic features indicate that the cave acted as a trap for the goats which inhabited the vicinity of the cave. These goats probably stumbled and fell to the bottom of the pit and they could not get out, dying inside. Due to the origin of the accumulation numerous complete bones have been recovered. These fossil allow us to perform a biometric analysis that indicates that population of Los Batanes is in the range size of other goats from Late Pleistocene of Iberian Peninsula

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

Measuring performance on the Healthcare Access and Quality Index for 195 countries and territories and selected subnational locations: A systematic analysis from the Global Burden of Disease Study 2016

Background: A key component of achieving universal health coverage is ensuring that all populations have access to quality health care. Examining where gains have occurred or progress has faltered across and within countries is crucial to guiding decisions and strategies for future improvement. We used the Global Burden of Diseases, Injuries, and Risk Factors Study 2016 (GBD 2016) to assess personal health-care access and quality with the Healthcare Access and Quality (HAQ) Index for 195 countries and territories, as well as subnational locations in seven countries, from 1990 to 2016. Methods Drawing from established methods and updated estimates from GBD 2016, we used 32 causes from which death should not occur in the presence of effective care to approximate personal health-care access and quality by location and over time. To better isolate potential effects of personal health-care access and quality from underlying risk factor patterns, we risk-standardised cause-specific deaths due to non-cancers by location-year, replacing the local joint exposure of environmental and behavioural risks with the global level of exposure. Supported by the expansion of cancer registry data in GBD 2016, we used mortality-to-incidence ratios for cancers instead of risk-standardised death rates to provide a stronger signal of the effects of personal health care and access on cancer survival. We transformed each cause to a scale of 0-100, with 0 as the first percentile (worst) observed between 1990 and 2016, and 100 as the 99th percentile (best); we set these thresholds at the country level, and then applied them to subnational locations. We applied a principal components analysis to construct the HAQ Index using all scaled cause values, providing an overall score of 0-100 of personal health-care access and quality by location over time. We then compared HAQ Index levels and trends by quintiles on the Socio-demographic Index (SDI), a summary measure of overall development. As derived from the broader GBD study and other data sources, we examined relationships between national HAQ Index scores and potential correlates of performance, such as total health spending per capita. Findings In 2016, HAQ Index performance spanned from a high of 97\ub71 (95% UI 95\ub78-98\ub71) in Iceland, followed by 96\ub76 (94\ub79-97\ub79) in Norway and 96\ub71 (94\ub75-97\ub73) in the Netherlands, to values as low as 18\ub76 (13\ub71-24\ub74) in the Central African Republic, 19\ub70 (14\ub73-23\ub77) in Somalia, and 23\ub74 (20\ub72-26\ub78) in Guinea-Bissau. The pace of progress achieved between 1990 and 2016 varied, with markedly faster improvements occurring between 2000 and 2016 for many countries in sub-Saharan Africa and southeast Asia, whereas several countries in Latin America and elsewhere saw progress stagnate after experiencing considerable advances in the HAQ Index between 1990 and 2000. Striking subnational disparities emerged in personal health-care access and quality, with China and India having particularly large gaps between locations with the highest and lowest scores in 2016. In China, performance ranged from 91\ub75 (89\ub71-93\ub76) in Beijing to 48\ub70 (43\ub74-53\ub72) in Tibet (a 43\ub75-point difference), while India saw a 30\ub78-point disparity, from 64\ub78 (59\ub76-68\ub78) in Goa to 34\ub70 (30\ub73-38\ub71) in Assam. Japan recorded the smallest range in subnational HAQ performance in 2016 (a 4\ub78-point difference), whereas differences between subnational locations with the highest and lowest HAQ Index values were more than two times as high for the USA and three times as high for England. State-level gaps in the HAQ Index in Mexico somewhat narrowed from 1990 to 2016 (from a 20\ub79-point to 17\ub70-point difference), whereas in Brazil, disparities slightly increased across states during this time (a 17\ub72-point to 20\ub74-point difference). Performance on the HAQ Index showed strong linkages to overall development, with high and high-middle SDI countries generally having higher scores and faster gains for non-communicable diseases. Nonetheless, countries across the development spectrum saw substantial gains in some key health service areas from 2000 to 2016, most notably vaccine-preventable diseases. Overall, national performance on the HAQ Index was positively associated with higher levels of total health spending per capita, as well as health systems inputs, but these relationships were quite heterogeneous, particularly among low-to-middle SDI countries. Interpretation GBD 2016 provides a more detailed understanding of past success and current challenges in improving personal health-care access and quality worldwide. Despite substantial gains since 2000, many low-SDI and middle- SDI countries face considerable challenges unless heightened policy action and investments focus on advancing access to and quality of health care across key health services, especially non-communicable diseases. Stagnating or minimal improvements experienced by several low-middle to high-middle SDI countries could reflect the complexities of re-orienting both primary and secondary health-care services beyond the more limited foci of the Millennium Development Goals. Alongside initiatives to strengthen public health programmes, the pursuit of universal health coverage hinges upon improving both access and quality worldwide, and thus requires adopting a more comprehensive view-and subsequent provision-of quality health care for all populations