NAC61 regulates late- and post-ripening osmotic, oxidative, and biotic stress responses in grapevine

During late- and post-ripening stages, grape berry undergoes profound biochemical and physiological changes whose molecular control is poorly understood. Here, we report the role of NAC61, a grapevine NAC transcription factor, in regulating different processes involved in berry ripening progression. NAC61 is highly expressed during post-harvest berry dehydration and its expression pattern is closely related to sugar concentration. The ectopic expression of NAC61 in Nicotiana benthamiana leaves resulted in low stomatal conductance, high leaf temperature, tissue collapse and a higher relative water content. Transcriptome analysis of grapevine leaves transiently overexpressing NAC61 and DNA affinity purification and sequencing analyses allowed us to narrow down a list of NAC61-regulated genes. Direct regulation of the stilbene synthase regulator MYB14, the osmotic stress-related gene DHN1b, the Botrytis cinerea susceptibility gene WRKY52, and NAC61 itself was validated. We also demonstrate that NAC61 interacts with NAC60, a proposed master regulator of grapevine organ maturation, in the activation of MYB14 and NAC61 expression. Overall, our findings establish NAC61 as a key player in a regulatory network that governs stilbenoid metabolism and osmotic, oxidative, and biotic stress responses that are the hallmark of late- and post-ripening grape stages.NAC61 regulates stilbene biosynthesis and abiotic/biotic stress responses that hallmark late- and post-ripening developmental stages in grapes. NAC61 self-activates and synergistically interacts with the master ripening regulator NAC60

Plant Signals Anticipate the Induction of the Type III Secretion System in Pseudomonas syringae pv. actinidiae, Facilitating Efficient Temperature-Dependent Effector Translocation

Disease resistance in plants depends on a molecular dialogue with microbes that involves many known chemical effectors, but the time course of the interaction and the influence of the environment are largely unknown. The outcome of host-pathogen interactions is thought to reflect the offensive and defensive capabilities of both players. When plants interact with Pseudomonas syringae, several well-characterized virulence factors contribute to early bacterial pathogenicity, including the type III secretion system (T3SS), which must be activated by signals from the plant and environment to allow the secretion of virulence effectors. The manner in which these signals regulate T3SS activity is still unclear. Here, we strengthen the paradigm of the plant-pathogen molecular dialogue by addressing overlooked details concerning the timing of interactions, specifically the role of plant signals and temperature on the regulation of bacterial virulence during the first few hours of the interaction. Whole-genome expression profiling after 1 h revealed that the perception of plant signals from kiwifruit or tomato extracts anticipated T3SS expression in P. syringae pv. actinidiae compared to apoplast-like conditions, facilitating more efficient effector transport in planta, as revealed by the induction of a temperature-dependent hypersensitive response in the nonhost plant Arabidopsis thaliana Columbia-0 (Col-0). Our results show that in the arms race between plants and bacteria, the temperature-dependent timing of bacterial virulence versus the induction of plant defenses is probably one of the fundamental parameters governing the outcome of the interaction. IMPORTANCE Plant diseases-their occurrence and severity-result from the impact of three factors: the host, the pathogen, and the environmental conditions, interconnected in the disease triangle. Time was further included as a fourth factor accounting for plant disease, leading to a more realistic three-dimensional disease pyramid to represent the evolution of disease over time. However, this representation still considers time only as a parameter determining when and to what extent a disease will occur, at a scale from days to months. Here, we show that time is a factor regulating the arms race between plants and pathogens, at a scale from minutes to hours, and strictly depends on environmental factors. Thus, besides the arms possessed by pathogens and plants per se, the opportunity and the timing of arms mobilization make the difference in determining the outcome of an interaction and thus the occurrence of plant disease

Guidelines for the use and interpretation of diagnostic methods in adult food allergy

Food allergy has an increasing prevalence in the general population and in Italy concerns 8 % of people with allergies. The spectrum of its clinical manifestations ranges from mild symptoms up to potentially fatal anaphylactic shock. A number of patients can be diagnosed easily by the use of first- and second-level procedures (history, skin tests and allergen specific IgE). Patients with complex presentation, such as multiple sensitizations and pollen-food syndromes, frequently require a third-level approach including molecular diagnostics, which enables the design of a component-resolved sensitization profile for each patient. The use of such techniques involves specialists' and experts' skills on the issue to appropriately meet the diagnostic and therapeutic needs of patients. Particularly, educational programs for allergists on the use and interpretation of molecular diagnostics are needed

ECMO for COVID-19 patients in Europe and Israel

Since March 15th, 2020, 177 centres from Europe and Israel have joined the study, routinely reporting on the ECMO support they provide to COVID-19 patients. The mean annual number of cases treated with ECMO in the participating centres before the pandemic (2019) was 55. The number of COVID-19 patients has increased rapidly each week reaching 1531 treated patients as of September 14th. The greatest number of cases has been reported from France (n = 385), UK (n = 193), Germany (n = 176), Spain (n = 166), and Italy (n = 136) .The mean age of treated patients was 52.6 years (range 16–80), 79% were male. The ECMO configuration used was VV in 91% of cases, VA in 5% and other in 4%. The mean PaO2 before ECMO implantation was 65 mmHg. The mean duration of ECMO support thus far has been 18 days and the mean ICU length of stay of these patients was 33 days. As of the 14th September, overall 841 patients have been weaned from ECMO support, 601 died during ECMO support, 71 died after withdrawal of ECMO, 79 are still receiving ECMO support and for 10 patients status n.a. . Our preliminary data suggest that patients placed on ECMO with severe refractory respiratory or cardiac failure secondary to COVID-19 have a reasonable (55%) chance of survival. Further extensive data analysis is expected to provide invaluable information on the demographics, severity of illness, indications and different ECMO management strategies in these patients

eCPMV NANOPARTICLES: THE POTENTIAL OF A BIO-INSPIRED STRATEGY FOR PLANT PROTECTION

Agricultural yield is dramatically affected by crop pests and pathogens, which are considered a major threat to food security. The intense and indiscriminate use of pesticides led to an increase in resistance occurrence in microorganisms and environmental issues, leading to the awareness towards the need for new alternative strategies respectful of both health and environment. Nanotechnology is one of the most promising technologies of the modern era. Indeed, its multidisciplinary nature makes it fascinating and exploitable in many fields, precisely including agriculture. Nanoagrochemicals can indeed provide a valuable alternative to common pesticides, due to a more targeted and enhanced efficacy, resulting in a lower dosage of active substances and a reduction of possible off-target effects. Although the biosafety of nanoagrochemicals is still not completely understood, their development for plant protection represents a promising strategy to achieve a sustainable and environmentally friendly crop production system. Plant viruses can play an important role in this context since they are naturally occurring proteinaceous nanoparticles able to carry, protect, and deliver a cargo. Moreover, the biosafety concerns about the use of plant viruses for crop protection have been exceeded by the development of nucleic acid-free virus nanoparticles, named empty virus-like particles (eVLPs). The Cowpea mosaic virus (CPMV) can be produced as eVLPs (eCPMV) through transient expression in Nicotiana benthamiana plants, at high titres and in an inexpensive manner. Additionally, eCPMV can be functionalized with peptides that can be displayed on the external surface by genetic manipulation or chemical conjugation, thus offering a wide range of possibilities for their application in plant protection. In this work, eCPMV nanoparticles have been exploited in an attempt to develop new products applicable for crop protection, using different approaches that rely on the physical stability of the eCPMV capsid to expose natural bioactive peptides or on the capacity of the empty eCPMV cavity to load natural molecules: (i) eCPMV as a scaffold for antimicrobial peptide (AMP) exposure, (ii), eCPMV as a plant immunity-triggering nanoparticle, and (iii) eCPMV as a functional nanocarrier.While it has not been possible to produce eCPMV harbouring AMPs on the surface, eCPMV functionalized with different elicitors were successfully expressed and purified and showed the capacity to induce plant molecular defence responses. Moreover, eCPMV nanoparticles were loaded with different molecules, efficiently further released in apoplastic-mimicking conditions, thus establishing the possibility for the cargo to be delivered within plant tissues. Overall, the data enforce to reconsider the paradigms regarding eCPMV functionalization, in particular in terms of peptide features required for genetical modification. Moreover, they support that eCPMV is a promising tool to develop new nanobiopesticides applicable in agriculture. Finally, they unveil unexpected effects of eCPMV nanoparticles, such as their capacity to disturb fungal growth and to fully protect plant from bacterial infection, which will certainly deserve attention for future researc

Temperature plays a role in the ability of Pseudomonas syringae strains carrying a recognized avirulence gene to induce a hypersensitive response in Arabidopsis thaliana

International audienc

MIRROR: Methodological Innovation to Remodel the Electric Loads to Reduce Economic OR Environmental Impact of User

Demand for electricity is constantly increasing, and production is facing new constraints due to the current world situation. An alternative to standard energy production methodologies is based on the use of renewable sources; however, these methodologies do not produce energy consistently due to weather factors. This results in a significant commitment of the user who must appropriately distribute loads in the most productive time slots. In this paper, a comparison is made between two methods of predicting solar energy production, one statistical and the other meteorological. For this work, a system capable of presenting the scheduling of household appliances is tested. The system is able to predict the energy consumption of the users and the energy production of the solar system. The system is tested using data from three different users, and the mean percentage of consumption reduction is about 77.73%. This is achieved through optimized programming of appliance use that also considers user comfort

Identification, characterization, and expression analysis of calmodulin and calmodulin-like genes in grapevine (Vitis vinifera) reveal likely roles in stress responses

Calcium (Ca2+) is an ubiquitous key second messenger in plants, where it modulates many developmental and adaptive processes in response to various stimuli. Several proteins containing Ca2+ binding domain have been identified in plants, including calmodulin (CaM) and calmodulin-like (CML) proteins, which play critical roles in translating Ca2+ signals into proper cellular responses. In this work, a genome-wide analysis conducted in Vitis vinifera identified three CaM- and 62 CML-encoding genes. We assigned gene family nomenclature, analyzed gene structure, chromosomal location and gene duplication, as well as protein motif organization. The phylogenetic clustering revealed a total of eight subgroups, including one unique clade of VviCaMs distinct from VviCMLs. VviCaMs were found to contain four EF-hand motifs whereas VviCML proteins have one to five. Most of grapevine CML genes were intronless, while VviCaMs were intron rich. All the genes were well spread among the 19 grapevine chromosomes and displayed a high level of duplication. The expression profiling of VviCaM/VviCML genes revealed a broad expression pattern across all grape organs and tissues at various developmental stages, and a significant modulation in biotic stress-related responses. Our results highlight the complexity of CaM/CML protein family also in grapevine, supporting the versatile role of its different members in modulating cellular responses to various stimuli, in particular to biotic stresses. This work lays the foundation for further functional and structural studies on specific grapevine CaMs/CMLs in order to better understand the role of Ca2+-binding proteins in grapevine and to explore their potential for further biotechnological applications

Isolated sphenoid inflammatory diseases associated with visual impairment: 15-year experience at a single institution

We present a retrospective series of 11 patients treated for isolated sphenoid inflammatory disease (ISID) with visual impairment in the period between 1994 and 2008. The series included 7 females and 4 males. All patients, preoperatively assessed by CT or MR, underwent endoscopic sinus surgery and broad-spectrum antibiotic therapy. The procedure was always performed in an emergency setting, with an interval from the onset of visual impairment ranging between 1 and 40 days (mean 9 days). The possibility of recovery was related to both the modality of onset and severity of the deficit. All patients with reduction of the visual field reported significant improvement after surgery. Patients with decreased visual acuity obtained partial or complete resolution, while in patients with preoperative blindness no improvement was observed. Moreover, no postoperative improvement was noticed in the case of severe deficits with sudden onset, whereas the treatment of mild deficits was successful even some weeks after their occurrence. In conclusion, although some factors may predict the likelihood of recovery, any patient with ISID associated with visual impairment should receive immediate medical and surgical treatment. Endoscopic surgery should be considered the technique of choice

Low-Cost Handheld Spectrometry for Detecting Flavescence Dorée in Vineyards

This study was conducted to evaluate the potential of low-cost hyperspectral sensors for the early detection of Flavescence dorée (FD) from asymptomatic samples prior to symptom development. In total, 180 leaf spectra from 60 randomly selected plants (three leaves per plant) were collected by using two portable mini-spectrometers (Hamamatsu: 340–850 nm and NIRScan: 900–1700 nm) at five vegetative growth stages in a vineyard with grape variety Garganega. High differences in the Hamamatsu spectra of the two groups were found in the VIS-NIR (visible–near infrared) spectral region while very small differences were observed in the NIRScan spectra. We analyzed the spectral data of two sensors by using all bands, features reduced by an ensemble method, and by genetic algorithms (GA) to discriminate the asymptomatic healthy (FD negative) and diseased (FD positive) leaves using five different classifiers. Overall, high classification accuracies were found in case of the Hamamatsu sensor compared to the NIRScan sensor. The feature selection techniques performed better compared to all bands, and the highest classification accuracy of 96% was achieved when GA features of the Hamamatsu sensor were used with the logistic regression (LR) classifier on test samples. A slightly low accuracy of 85% was achieved when the features (selected by the ensemble method) of the Hamamatsu sensor were used with the support vector machine (SVM) classifier by using leave-one-out (LOO) cross-validation on the whole dataset. Results demonstrated that employing a feature selection technique can provide a valid tool for determining the optimal bands that can be used to identify FD disease in the vineyard. However, further validation studies are required, as this study was conducted using a small dataset and from the single grapevine variety