Carbon budget of the vineyard \u2013 A new feature of sustainability

Vineyards received scarce attention in relation to the continuous monitoring of carbon fluxes and the assessment of their overall budget, as a common believe is that agricultural crops cannot be net carbon sinks. Indeed, many technical inputs, massive periodical harvests, and the repeated disturbances of upper soil layers, all contribute to a substantial loss both of the old and newly-synthesized organic matter. Woody perennials, however, can behave differently: they grow a permanent structure, stand undisturbed in the same field for decades, originate abundant pruning debris, and are often grass-covered. We have been monitoring the Net Ecosystem Exchange (NEE) by eddy covariance and the carbon partitioning in a temperate vineyard in North Eastern Italy. Five complete yearly budgets confirm a steady and substantial sink capacity of the system, with a yearly NEE around 800\u2013900 gC m 122, grape harvest representing about 20\u201325% of it. Biometrical assessment of growth and partitioning show a good agreement with micrometeorological measurements and demonstrate a large input of organic matter into the soil. Even if it can be objected that this sink may be only temporary and the built-up can be substantially disrupted at the end of the vineyard life cycle, these results show that there is a concrete possibility of storing carbon in temperate-climate vineyards, possibly contributing to the global carbon budget. This sink capacity might be accounted in the official calculation of wine carbon footprint and represents a new, relevant feature of their sustainability

Sentinel-2 Data Analysis and Comparison with UAV Multispectral Images for Precision Viticulture

Precision viticulture (PV) requires the use of technologies that can detect the spatial and temporal variability of vineyards and, at the same time, allow useful information to be obtained at sustainable costs. In order to develop a cheap and easy-to-handle operational monitoring scheme for PV, the aim of this work was to evaluate the possibility of using Sentinel-2 multispectral images for long-term vineyard monitoring through the Normalized Difference Vegetation Index (NDVI). Vigour maps of two vineyards located in northeastern Italy were computed from satellite imagery and compared with those derived from UAV multispectral images; their correspondence was evaluated from qualitative and statistical points of view. To achieve this, the UAV images were roughly resampled to 10 m pixel size in order to match the spatial resolution of the satellite imagery. Preliminary results show the potential use of open source Sentinel-2 platforms for monitoring vineyards, highlighting links with the information given in the agronomic bulletins and identifying critical areas for crop production. Despite the large differences in spatial resolution, the results of the comparison between the UAV and Sentinel-2 data were promising. However, for long-term vineyard monitoring at territory scale, further studies using multispectral sensor calibration and groundtruth data are required

Transcriptome pathways in leaf and root of grapevine genotypes with contrasting drought tolerance

Most of the world’s wine-producing regions are subjected to seasonal drought,and,in the light of the dramatic climate-change events occurring in recent years, the selection of resistant rootstocks is becoming a crucial factor for the development of sustainable agricultural models to ensure optimal grape berry development and ripening. In this study, roots and leaves of 101.14 (drought-susceptible) and M4 (drought-tolerant) rootstocks were sampled in progressive drought and mRNA-seq profiles were evaluated. Physiological characterization indicated that only M4 was able to maintain high leaf transpiration and net assimilation rates under severe stress conditions. Statistical analyses, carried out on mRNA-seq data, highlighted that “treatment” (water stress) and “genotype” (rootstock-genotype) seem to be the main variables explaining differential gene expression in roots and leaves tissues, respectively. Upon water-stress, roots and leaves of the tolerant genotype M4 exhibit a higher induction of stilbenes (i.e., STS) and flavonoids (e.g., CHS, F3H, FLS) biosynthetic genes. Moreover, the higher expression of STS genes in M4 is coupled with an up-regulation of WRKYs transcription factors. STS genes promoter regions, extracted from whole genome of M4 and 101.14, highlighted a higher number of WBOX cis elements (binding site for WRKYs) in the tolerant genotype

WAVE2 is regulated by multiple phosphorylation events within its VCA domain.

addresses: Department of Biochemistry, Faculty of Medical and Veterinary Sciences, University of Bristol, Bristol, United Kingdom.notes: PMCID: PMC2798068OnlineOpen Article. This is a copy of an article published in Cell Motility and the Cytoskeleton. This journal is available online at: http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1949-3592The definitive version is available at www3.interscience.wiley.comThe (Wiskott-Aldrich Syndrome Protein)-family verprolin homologous protein (WAVE) family of proteins occupies a pivotal position in the cell, converting extracellular signals into the formation of branched filamentous (F) actin structures. WAVE proteins contain a verprolin central acidic (VCA) domain at their C-terminus, responsible for binding to and activating the Arp2/3 complex, which in-turn nucleates the formation of new actin filaments. Here we identify five Casein Kinase 2 (CK2) phosphorylation sites within the VCA domain of WAVE2, serines 482, 484, 488, 489, and 497. Phosphorylation of these sites is required for a high affinity interaction with the Arp2/3 complex. Phosphorylation of ser 482 and 484 specifically inhibits the activation of the Arp2/3 complex by the WAVE2 VCA domain, but has no effect on the affinity for the Arp2/3 complex when the other phosphorylation sites are occupied. We demonstrate phosphorylation of all five sites on endogenous WAVE2 and show that their mutation to non-phosphorylatable alanine residues inhibits WAVE2 function in vivo, inhibiting cell ruffling and disrupting the integrity of the leading edge of migrating cells

Grapevine Rootstocks Differentially Affect the Rate of Ripening and Modulate Auxin-Related Genes in Cabernet Sauvignon Berries

In modern viticulture, grafting commercial grapevine varieties on interspecific rootstocks is a common practice required for conferring resistance to many biotic and abiotic stresses. Nevertheless, the use of rootstocks to gain these essential traits is also known to impact grape berry development and quality, although the underlying mechanisms are still poorly understood. In grape berries, the onset of ripening (v\ue9raison) is regulated by a complex network of mobile signals including hormones such as auxins, ethylene, abscisic acid, and brassinosteroids. Recently, a new rootstock, designated M4, was selected based on its enhanced tolerance to water stress and medium vigor. This study investigates the effect of M4 on Cabernet Sauvignon (CS) berry development in comparison to the commercial 1103P rootstock. Physical and biochemical parameters showed that the ripening rate of CS berries is faster when grafted onto M4. A multifactorial analysis performed on mRNA-Seq data obtained from skin and pulp of berries grown in both graft combinations revealed that genes controlling auxin action (ARF and Aux/IAA) represent one of main categories affected by the rootstock genotype. Considering that the level of auxin tightly regulates the transcription of these genes, we investigated the behavior of the main gene families involved in auxin biosynthesis and conjugation. Molecular and biochemical analyses confirmed a link between the rate of berry development and the modulation of auxin metabolism. Moreover, the data indicate that this phenomenon appears to be particularly pronounced in skin tissue in comparison to the flesh

A multicentre case control study on complicated coeliac disease: two different patterns of natural history, two different prognoses.

Abstract Background: Coeliac disease is a common enteropathy characterized by an increased mortality mainly due to its complications. The natural history of complicated coeliac disease is characterised by two different types of course: patients with a new diagnosis of coeliac disease that do not improve despite a strict gluten-free diet (type A cases) and previously diagnosed coeliac patients that initially improved on a gluten-free diet but then relapsed despite a strict diet (type B cases). Our aim was to study the prognosis and survival of A and B cases. Methods: Clinical and laboratory data from coeliac patients who later developed complications (A and B cases) and sex- and age-matched coeliac patients who normally responded to a gluten-free diet (controls) were collected among 11 Italian centres. Results: 87 cases and 136 controls were enrolled. Complications tended to occur rapidly after the diagnosis of coeliac disease and cumulative survival dropped in the first months after diagnosis of complicated coeliac disease. Thirty-seven cases died (30/59 in group A, 7/28 in group B). Type B cases presented an increased survival rate compared to A cases. Conclusions: Complicated coeliac disease is an extremely serious condition with a high mortality and a short survival. Survival depends on the type of natural history. Keyword: Celiac disease, Complications, EATL, Prognosis, Glutens, Gluten-free die

Medium-resolution multispectral data from sentinel-2 to assess the damage and the recovery time of late frost on Vineyards

In a climate-change context, the advancement of phenological stages may endanger viticultural areas in the event of a late frost. This study evaluated the potential of satellite-based remote sensing to assess the damage and the recovery time after a late frost event in 2017 in northern Italian vineyards. Several vegetation indices (VIs) normalized on a two-year dataset (2018-2019) were compared over a frost-affected area (F) and a control area (NF) using unpaired two-sample t-test. Furthermore, the must quality data (total acidity, sugar content and pH) of F and NF were analyzed. The VIs most sensitive in the detection of frost damage were Chlorophyll Absorption Ratio Index (CARI), Enhanced Vegetation Index (EVI), and Modified Triangular Vegetation Index 1 (MTVI1) (-5.26%,-16.59%, and-5.77% compared to NF, respectively). The spectral bands Near-Infrared (NIR) and Red Edge 7 were able to identify the frost damage (-16.55 and-16.67% compared to NF, respectively). Moreover, CARI, EVI, MTVI1, NIR, Red Edge 7, the Normalized Difference Vegetation Index (NDVI) and the Modified Simple Ratio (MSR) provided precise information on the full recovery time (+17.7%, +22.42%, +29.67%, +5.89%, +5.91%, +16.48%, and +8.73% compared to NF, respectively) approximately 40 days after the frost event. The must analysis showed that total acidity was higher (+5.98%), and pH was lower (-2.47%) in F compared to NF. These results suggest that medium-resolution multispectral data from Sentinel-2 constellation may represent a cost-effective tool for frost damage assessment and recovery management

Metformin attenuates the effect of Staphylococcus aureus on airway tight junctions by increasing PKCζ-mediated phosphorylation of occludin.

Airway epithelial tight junction (TJ) proteins form a resistive barrier to the external environment, however, during respiratory bacterial infection TJs become disrupted compromising barrier function. This promotes glucose flux/accumulation into the lumen which acts as a nutrient source for bacterial growth. Metformin used for the treatment of diabetes increases transepithelial resistance (TEER) and partially prevents the effect of bacteria but the mechanisms of action are unclear. We investigated the effect of metformin and Staphylococcus aureus on TJ proteins, zonula occludins (ZO)-1 and occludin in human airway epithelial cells (H441). We also explored the role of AMP-activated protein kinase (AMPK) and PKCζ in metformin-induced effects. Pretreatment with metformin prevented the S. aureus-induced changes in ZO-1 and occludin. Metformin also promoted increased abundance of full length over smaller cleaved occludin proteins. The nonspecific PKC inhibitor staurosporine reduced TEER but did not prevent the effect of metformin indicating that the pathway may involve atypical PKC isoforms. Investigation of TJ reassembly after calcium depletion showed that metformin increased TEER more rapidly and promoted the abundance and localization of occludin at the TJ. These effects were inhibited by the AMPK inhibitor, compound C and the PKCζ pseudosubstrate inhibitor (PSI). Metformin increased phosphorylation of occludin and acetyl-coA-carboxylase but only the former was prevented by PSI. This study demonstrates that metformin improves TJ barrier function by promoting the abundance and assembly of full length occludin at the TJ and that this process involves phosphorylation of the protein via an AMPK-PKCζ pathway

Utilizing GIS tools to analyze viticultural choices under climate change scenario in North-East of Italy

Vineyard areas are constantly decreasing in Italy as well as in Europe. North-eastern regions in Italy are showing an opposite trend, steadily expanding with increased winegrowing areas. In viticulture and wine production, climate is arguably the most critical aspect in ripening fruit to achieve optimum characteristics to produce a given wine style. According to WMO and IPCC, climate is changing and the world is experiencing unprecedented climate extremes. Despite recent zoning aimed at defining key factors in determining the suitability of a given region for specific varieties and wine types, the expansion of viticulture in North East of Italy has led to some irrational planting choices about row orientation, dimensions, and slope. Under these conditions, the consequences of some extreme weather events may be more severe. The main objective of this study was to verify whether row orientation, aspect, and slope of vineyards, in combination with climate conditions, may affect yield and fruit quality. An area localized in the Northern Italy was analyzed, taking advantage of QGIS tools. The investigated parameters included: row orientation, slope, area, age of plantation, aspect ratio and distance between rows. Such variables have been combined with management information (planting distances, scion/rootstock combination, use of irrigation) and environmental information (yearly weather conditions). Data resulting from GIS analysis, vineyard management and environmental information have been correlated with 10-years yield and must quality parameters. Furthermore, satellite imagery from sample vineyards were collected and investigated in order to analyze the responses of the plants to different weather conditions. The results of the analysis highlighted how the mean slope of investigated vineyards is in general ranging between 1 and 3 degrees, with a prevalent Southern exposure. Rows do not exhibit a dominant orientation, mainly due to the following reasons: - the reduced dimensions available for vine cultivation, especially in hilly areas, where the vineyards are planted along contours, in order to limit erosion - the need for mechanisation, which calls for longer rather than larger rows. The results enabled to create a connection between row orientation, climate and soil conditions, and grapevine yield and quality responses to be considered as a guide for future planting choices more suitable to the restrictions imposed by increasing extreme weather events

Protein Kinase CK2 Mutants Defective in Substrate Recognition PURIFICATION AND KINETIC ANALYSIS

Five mutants of protein kinase CK2 α subunit in which altogether 14 basic residues were singly to quadruply replaced by alanines (K74A,K75A,K76A,K77A; K79A, R80A,K83A; R191A,R195A,K198A; R228A; and R278A, K279A,R280A) have been purified to near homogeneity either as such or after addition of the recombinant β subunit. By this latter procedure five mutated tetrameric holoenzymes were obtained as judged from their subunit composition, sedimentation coefficient on sucrose gradient ultracentrifugation, and increased activity toward a specific peptide substrate as compared with the isolated α subunits. The kinetic constants and the phosphorylation efficiencies (Vmax/Km) of all the mutants with the parent peptide RRRADDSDDDDD and a series of derivatives, in which individual aspartic acids were replaced by alanines, have been determined. Three mutants, namely K74A,K75A,K76A,K77A; K79A,R80A, K83A; and R191A,R195A,K198A display dramatically lower phosphorylation efficiency and 8-50-fold higher Km values with the parent peptide, symptomatic of reduced attitude to bind the peptide substrate as compared with CK2 wild type. Such differences either disappear or are attenuated if the mutants R191A,R195A, K198A; K79A,R80A,K83A; and K74A,K75A,K76A,K77A are assayed with the peptides RRRADDSADDDD, RRRADDSDDADD, and RRRADDSDDDAA, respectively. In contrast, the phosphorylation efficiencies of the other substituted peptides decrease more markedly with these mutants than with CK2 wild type. These data show that one or more of the basic residues clustered in the 191-198, 79-83, and 74-77 sequences are implicated in the recognition of the acidic determinants at positions +1, +3, and +4/+5, respectively, and that if these residues are mutated, the relevance of the other acidic residues surrounding serine is increased. In contrast the other two mutants, namely R228A and R278A,K279A, R280A, display with all the peptides Vmax values higher than CK2 wild type, counterbalanced however by somewhat higher Kmvalues. It can be concluded from these data that all the five mutations performed are compatible with the reconstitution of tetrameric holoenzyme, but all of them influence the enzymatic efficiency of CK2 to different extents. Although the basic residues mutated in the 74-77, 79-83, and 191-198 sequences are clearly implicated in substrate recognition by interacting with acidic determinants at variable positions downstream from serine, the other basic residues seem to play a more elusive and/or indirect role in catalysis