Genomics of osmotic stress responses in Arabidopsis and crops

Drought and soil salinity limit available land for agriculture and reduce crop yields by imposing osmotic stress on plants. The identification of genes involved in plant tolerance mechanisms is a key goal to develop crops better capable to handle these stresses. The aim of this project was to identify genes involved in the plant response to osmotic stress using genomic approaches both in the model plant Arabidopsis thaliana and in a major crop in the Mediterranean basin, Solanum lycopersicum. Previously, several genes whose expression was differentially regulated in Solanum tuberosum culture cells adapted to increasing concentrations of polyethylene glycol (PEG) were identified. Here, the functional role of fifty of these genes was verified by studying their orthologues in A. thaliana. Homozygous knockout lines for each gene were subjected to a large-scale phenotype screening in order to identify genes involved in adaptation to osmotic stress. Using this strategy, we have identified 3 genes whose function in stress response was so far unknown: the splicing factor DRT111, the putative negative regulator of TOR pathway TIP41-like, the subunit of Pol III SIN-like. A detailed functional characterization for these three selected genes was performed through the use of specific advanced molecular technologies. Interestingly, germination analysis on plants with alterated expression of DRT111 as well as the reported association with another important splicing factor SUA suggest that DRT111 is involved in pre-mRNA splicing of ABI3, regulating ABA-related inhibition of seed germination. Similarly, TIP41-like possibly affects several pathways in the ABA-mediated response to osmotic stress. The abolished expression of TIP41-like leads to ABA hypersensitive phenotypes at germination and seedling stage as well as reduction in root development, indicating alteration in ABA biosynthesis and/or perception. Finally, plants with abolished expression of SIN-like show severe reduction in root growth, suggesting that the mutation in SIN-like leads to phenotypes mainly associated with altered cell development that could be explained by an alteration of RNAs transcribed by Pol III. Growing evidence, indeed, demonstrates that RNA regulatory mechanisms participate in the modulation of abiotic stress responses. RNA binding proteins play an important role in such mechanisms, but relatively few have been characterized in plants so far. Among the differentially expressed genes in S.tuberosum adapted cells, the Arabidopsis ortholog AtRGGA, encoding a glycine-rich RNA-binding protein, was previously partially characterized in responses to drought and salt stress. Here we analyze the protein role in the RNA regulatory mechanisms. As expected from the presence of Arg/Gly motif, we provide evidence that AtRGGA is capable to binding ribosomal RNAs both in vitro and in vivo. To identify protein partners of AtRGGA, yeast two-hybrid screening was performed using an Arabidopsis cDNA library. Most of the identified proteins are involved in RNA processing, transport and ribosome biogenesis. Therefore, the obtained results indicate a role of AtRGGA in transcriptional and post-transcriptional control of gene expression during osmotic stress. Furthermore, this study also gives a complete picture of the mechanisms employed to maintain a cellular and whole-plant homeostasis in tomato during drought stress. Transcriptome profile was analyzed in M82 genotype during two cycles of prolonged drought stress and one of rewatering, monitoring physiological parameters as well as ABA and proline accumulation. The observed reduction in leaf gas exchanges was concomitant with a general arrest of transcriptional activities. In particular, Gene Onthology (GO) categories such as cell proliferation and cell cycle were significantly enriched in the down-regulated fraction of genes upon drought stress. As expected, ABA and proline accumulated after prolonged water deficit, driving the observed enrichment of stress related GOs in the up-regulated gene fractions, which included transcripts putatively involved in stomatal movements as well as cellular homeostasis maintenance. Taken together, our results revealed promising mechanisms of plant acclimation to stress. DRT111, TIP41-like, SIN-like and AtRGGA are active participants in the response to drought and salinity and represent interesting targets for future use in crop species. Furthermore, the analyzed interconnection between the physiological responses and gene expression in tomato has given other interesting candidate genes that could play novel roles in drought tolerance and adaptation

Transcriptomic and splicing changes underlying tomato responses to combined water and nutrient stress

Tomato is a horticultural crop of high economic and nutritional value. Suboptimal environmental conditions, such as limited water and nutrient availability, cause severe yield reductions. Thus, selection of genotypes requiring lower inputs is a goal for the tomato breeding sector. We screened 10 tomato varieties exposed to water deficit, low nitrate or a combination of both. Biometric, physiological and molecular analyses revealed different stress responses among genotypes, identifying T270 as severely affected, and T250 as tolerant to the stresses applied. Investigation of transcriptome changes caused by combined stress in roots and leaves of these two genotypes yielded a low number of differentially expressed genes (DEGs) in T250 compared to T270, suggesting that T250 tailors changes in gene expression to efficiently respond to combined stress. By contrast, the susceptible tomato activated approximately one thousand and two thousand genes in leaves and roots respectively, indicating a more generalized stress response in this genotype. In particular, developmental and stress-related genes were differentially expressed, such as hormone responsive factors and transcription factors. Analysis of differential alternative splicing (DAS) events showed that combined stress greatly affects the splicing landscape in both genotypes, highlighting the important role of AS in stress response mechanisms. In particular, several stress and growth-related genes as well as transcription and splicing factors were differentially spliced in both tissues. Taken together, these results reveal important insights into the transcriptional and post-transcriptional mechanisms regulating tomato adaptation to growth under reduced water and nitrogen inputs

Von Hippel-Lindau–dependent polycythemia is endemic on the island of Ischia: identification of a novel cluster

AbstractChuvash polycythemia (MIM 263400) is an autosomal recessive disorder characterized by a high hemoglobin level, relatively high serum erythropoietin, and early death. It results from a Von Hippel-Lindau (VHL) gene mutation (C598T) that causes increased HIF-1α activity and erythrocyte production in the face of normoxia. This polycythemia is endemic in Chuvashia, whereas its worldwide frequency is very low. We investigated the incidence of the Chuvash-type VHL mutation in Campania (South Italy) and identified 14 affected subjects (5 families). Twelve live on the island of Ischia (Bay of Naples). From analysis of the mutated allele, we found that the disease was more frequent on Ischia (0.070) than in Chuvashia (0.057). The haplotype of all patients matched that identified in the Chuvash cluster, thereby supporting the single-founder hypothesis. We also found that nonaffected heterozygotes had increased HIF-1α activity, which might confer a biochemical advantage for mutation maintenance. In conclusion, we have identified the first large cluster of Chuvash erythrocytosis outside Chuvashia, which suggests that this familial polycythemia might be endemic in other regions of the world

Fermentation of Vaccinium floribundum Berries with Lactiplantibacillus plantarum Reduces Oxidative Stress in Endothelial Cells and Modulates Macrophages Function

Accumulating evidence suggests that high consumption of natural antioxidants promotes health by reducing oxidative stress and, thus, the risk of developing cardiovascular diseases. Similarly, fermentation of natural compounds with lactic acid bacteria (LAB), such as Lactiplantibacillus plantarum, enhances their beneficial properties as regulators of the immune, digestive, and cardiovascular system. We investigated the effects of fermentation with Lactiplantibacillus plantarum on the antioxidant and immunomodulatory effects of Pushgay berries (Vaccinium floribundum, Ericaceae family) in human umbilical vein endothelial cells (HUVECs) and macrophage cell line RAW264.7. Polyphenol content was assayed by Folin-Ciocalteu and HPLC-MS/MS analysis. The effects of berries solutions on cell viability or proliferation were assessed by WST8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt and Lactate dehydrogenase (LDH) release, Trypan blue exclusion test, and Alamar blue assay. Antioxidant activity was evaluated by a cell-based chemiluminescent probe for the detection of intracellular H2O2 production in HUVECs. Heme oxygenase-1 (HO-1) expression levels were investigated by RT-qPCR. Glutathione reductase (GR), glutathione peroxidase (Gpx), superoxide dismutase (SOD), and catalase (CAT) activities, as markers of intracellular antioxidant defense, were evaluated by spectrophotometric analysis. The immunomodulatory activity was examined in RAW 264.7 by quantification of inducible nitric oxide synthase (iNOS) and Tumor Necrosis Factor-alpha (TNF alpha) by RT-qPCR. Data showed that fermentation of Pushgay berries (i) enhances the content of quercetin aglycone, and (ii) increases their intracellular antioxidant activity, as indicated by the reduction in H2O2-induced cell death and the decrease in H2O2-induced HO-1 gene expression in HUVECs treated for 24 h with fermented berries solution (10 mu g/mL). Moreover, treatment with Pushgay berries for 72 h (10 mu g/mL) promotes cells growth in RAW 264.7, and only fermented Pushgay berries increase the expression of iNOS in the same cell line. Taken together, our results show that LAB fermentation of Pushgay berries enhances their antioxidant and immunomodulatory properties

Salinity and ABA Seed Responses in Pepper: Expression and Interaction of ABA Core Signaling Components

Abscisic acid (ABA) plays an important role in various aspects of plant growth and development, including adaptation to stresses, fruit development and ripening. In seeds, ABA participates through its core signaling components in dormancy instauration, longevity determination, and inhibition of germination in unfavorable environmental conditions such as high soil salinity. Here, we show that seed germination in pepper was delayed but only marginally reduced by ABA or NaCl with respect to control treatments. Through a similarity search, pepper orthologs of ABA core signaling components PYL (PYRABACTIN RESISTANCE1-LIKE), PP2C (PROTEIN PHOSPHATASE2C), and SnRK2 (SUCROSE NONFERMENTING1 (SNF1)-RELATED PROTEIN KINASE2) genes were identified. Gene expression analyses of selected members showed a low abundance of PYL and SnRK2 transcripts in dry seeds compared to other tissues, and an up-regulation at high concentrations of ABA and/or NaCl for both positive and negative regulators of ABA signaling. As expected, in hydroponically-grown seedlings exposed to NaCl, only PP2C encoding genes were up-regulated. Yeast two hybrid assays performed among putative pepper core components and with Arabidopsis thaliana orthologs confirmed the ability of the identified proteins to function in ABA signaling cascade, with the exception of a CaABI isoform cloned from seeds. BiFC assay in planta confirmed some of the interactions obtained in yeast. Altogether, our results indicate that a low expression of perception and signaling components in pepper seeds might contribute to explain the observed high percentages of seed germination in the presence of ABA. These results might have direct implications on the improvement of seed longevity and vigor, a bottleneck in pepper breeding

Cardiac magnetic resonance predictors of left ventricular remodelling following acute ST elevation myocardial infarction: The VavirimS study

Left ventricular (LV) remodelling (REM) ensuing after ST-elevation myocardial infarction (STEMI), has typically been studied by echocardiography, which has limitations, or cardiac magnetic resonance (CMR) in early phase that may overestimate infarct size (IS) due to tissue edema and stunning. This prospective, multicenter study investigated LV-REM performing CMR in the subacute phase, and 6 months after STEMI

Monitoraggio integrato di un'area marino-costiera: la foce del fiume Volturno (Mar Tirreno centrale)

Vengono presentati i risultati dell’attività di monitoraggio svolta nel corso del progetto PONa3_00363 I-AMICA (Infrastruttura di Alta tecnologia per il Monitoraggio Integrato Climatico-Ambientale; www.i-amica.it/i-amica/), nell’ambito delle attività relative all’Obiettivo Realizzativo 4.4 (Processi di interfaccia biosfera-idrosfera e funzionalità degli ecosistemi costieri). L’attività, che ha avuto come scopo l’acquisizione di conoscenze avanzate sulle dinamiche e/o variazioni nel tempo degli ecosistemi marino-costieri in relazione ai processi fisici, chimici e biologici che caratterizzano il loro habitat, si è sviluppata attraverso la sperimentazione di nuove metodologie di monitoraggio in relazione alle specifiche caratteristiche dell’area marino-costiera prospiciente la foce del fiume Volturno (Golfo di Gaeta). In particolare, oltre che sulle tecniche di monitoraggio classiche, lo studio si è principalmente focalizzato sulla acquisizione ed interpretazione di dati ambientali sia in colonna d’acqua che nei sedimenti a fondo mare e sulla identificazione di specie e/o associazioni di specie significative (bio-indicatori) da un punto di vista ambientale ed indicative dello stato di salute del sistema costiero. Poiché il sistema costiero rappresenta una struttura naturale complessa e delicata, la cui evoluzione è il risultato di delicati equilibri fisici, chimici e biologici, fortemente condizionabili dagli interventi antropici, l’attività di monitoraggio è stata integrata da studi sulla variazione della linea di costa, da studi sismostratigrafici della piana deltizia, sedimentologici e morfo-batimetrici dei fondali.Published1-714A. Oceanografia e climaJCR Journa

Epidemiological characteristics of COVID-19 cases and estimates of the reproductive numbers 1 month into the epidemic, Italy, 28 January to 31 March 2020

BackgroundOn 20 February 2020, a locally acquired coronavirus disease (COVID-19) case was detected in Lombardy, Italy. This was the first signal of ongoing transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the country. The number of cases in Italy increased rapidly and the country became the first in Europe to experience a SARS-CoV-2 outbreak.AimOur aim was to describe the epidemiology and transmission dynamics of the first COVID-19 cases in Italy amid ongoing control measures.MethodsWe analysed all RT-PCR-confirmed COVID-19 cases reported to the national integrated surveillance system until 31 March 2020. We provide a descriptive epidemiological summary and estimate the basic and net reproductive numbers by region.ResultsOf the 98,716 cases of COVID-19 analysed, 9,512 were healthcare workers. Of the 10,943 reported COVID-19-associated deaths (crude case fatality ratio: 11.1%) 49.5% occurred in cases older than 80 years. Male sex and age were independent risk factors for COVID-19 death. Estimates of R0 varied between 2.50 (95% confidence interval (CI): 2.18-2.83) in Tuscany and 3.00 (95% CI: 2.68-3.33) in Lazio. The net reproduction number Rt in northern regions started decreasing immediately after the first detection.ConclusionThe COVID-19 outbreak in Italy showed a clustering onset similar to the one in Wuhan, China. R0 at 2.96 in Lombardy combined with delayed detection explains the high case load and rapid geographical spread. Overall, Rt in Italian regions showed early signs of decrease, with large diversity in incidence, supporting the importance of combined non-pharmacological control measures

Diversity and ethics in trauma and acute care surgery teams: results from an international survey

Background Investigating the context of trauma and acute care surgery, the article aims at understanding the factors that can enhance some ethical aspects, namely the importance of patient consent, the perceptiveness of the ethical role of the trauma leader, and the perceived importance of ethics as an educational subject. Methods The article employs an international questionnaire promoted by the World Society of Emergency Surgery. Results Through the analysis of 402 fully filled questionnaires by surgeons from 72 different countries, the three main ethical topics are investigated through the lens of gender, membership of an academic or non-academic institution, an official trauma team, and a diverse group. In general terms, results highlight greater attention paid by surgeons belonging to academic institutions, official trauma teams, and diverse groups. Conclusions Our results underline that some organizational factors (e.g., the fact that the team belongs to a university context or is more diverse) might lead to the development of a higher sensibility on ethical matters. Embracing cultural diversity forces trauma teams to deal with different mindsets. Organizations should, therefore, consider those elements in defining their organizational procedures. Level of evidence Trauma and acute care teams work under tremendous pressure and complex circumstances, with their members needing to make ethical decisions quickly. The international survey allowed to shed light on how team assembly decisions might represent an opportunity to coordinate team member actions and increase performance