The grapevine industry is of high economic importance in several countries worldwide. Its growing market demand led to acceleration of the whole production processes, implying increasing water resources use at the expense of environmental water balance and hydrological cycle. Furthermore, in recent decades climate change and the consequent expansion of drought have further compromised water availability, making current agricultural systems even more fragile from ecological and economical perspectives. Consequently, farmers' income and welfare are increasingly unpredictable and unstable. Therefore, it is urgent to improve the resilience of vineyards, and of agro-ecosystems in general, by developing sustainable and environmentally-friendly farming practices by more rational biological and natural resources use. The PRIMA project PROSIT addresses these challenges by characterizing and harnessing grapevine-associated microbiota to propose innovative and sustainable agronomic practices. In particular, PROSIT aims to determine the efficacy of natural microbiomes transferred from grapevines adapted to arid climate to commonly cultivated grapevine cultivars. In doing so it will test those natural microbiome effects on drought tolerance. This multidisciplinary project will thus utilize in vitro culture techniques, bioimaging, microbiological tests, metabolomics, metabarcoding and epigenetic analyses. These will be combined to shed light on molecular mechanisms triggered in plants by microbial associations upon water stress. To this end it is hoped that the project will serve as a blueprint not only for studies uncovering the microbiome role in drought stress in a wide range of species, but also for analyzing its effect on a wide range of stresses commonly encountered in modern agricultural systems.

The environmental impact and health of ruminants fed with forage legumes depend on the herbage's concentration and structure of proanthocyanidins (PAs). Unfortunately, the primary forage legumes (alfalfa and clover) do not contain substantial levels of PAs. No significant progress has been made to induce PAs to agronomically valuable levels in their edible organs by biotechnological approaches thus far. Building this trait requires a profound knowledge of PA regulators and their interplay in species naturally committed to accumulating these metabolites in the target organs. Against this background, we compared the shoot transcriptomes of two inter-fertile Lotus species, namely Lotus tenuis and Lotus corniculatus, polymorphic for this trait, to search for differentially expressed MYB and bHLH genes. We then tested the expression of the above-reported regulators in L. tenuis x L. corniculatus interspecific hybrids, several Lotus spp., and different L. corniculatus organs with contrasting PA levels. We identified a novel MYB activator and MYB-bHLH-based complexes that, when expressed in Nicotiana benthamiana, trans-activated the promoters of L. corniculatus anthocyanidin reductase and leucoanthocyanidin reductase 1 genes. The last are the two critical structural genes for the biosynthesis of PAs in Lotus spp. Competition between MYB activators for the transactivation of these promoters also emerged. Overall, by employing Lotus as a model genus, we refined the transcriptional network underlying PA biosynthesis in the herbage of legumes. These findings are crucial to engineering this trait in pasture legumes.

The future of biomaterial production will leverage biotechnology based on the domestication of cells as biological factories. Plants, algae, and bacteria can produce low-environmental impact biopolymers. Here, two strategies were developed to produce a biopolymer derived from a bioengineered vacuolar storage protein of the common bean (phaseolin; PHSL). The cys-added PHSL* forms linear-structured biopolymers when expressed in the thylakoids of transplastomic tobacco leaves by exploiting the formation of inter-chain disulfide bridges. The same protein without signal peptide (?PHSL*) accumulates in Escherichia coli inclusion bodies as high-molarmass species polymers that can subsequently be oxidized to form disulfide crosslinking bridges in order to increase the stiffness of the biomaterial, a valid alternative to the use of chemical crosslinkers. The E. coli cells produced 300 times more engineered PHSL, measured as percentage of total soluble proteins, than transplastomic tobacco plants. Moreover, the thiol groups of cysteine allow the site-specific PEGylation of ?PHSL*, which is a desirable functionality in the design of a protein-based drug carrier. In conclusion, ?PHSL* expressed in E. coli has the potential to become an innovative biopolymer.

Model species continue to underpin groundbreaking plant science research. At the same time, the phylogenetic resolution of the land plant tree of life continues to improve. The intersection of these 2 research paths creates a unique opportunity to further extend the usefulness of model species across larger taxonomic groups. Here we promote the utility of the Arabidopsis thaliana model species, especially the ability to connect its genetic and functional resources, to species across the entire Brassicales order. We focus on the utility of using genomics and phylogenomics to bridge the evolution and diversification of several traits across the Brassicales to the resources in Arabidopsis, thereby extending scope from a model species by establishing a "model clade." These Brassicales-wide traits are discussed in the context of both the model species Arabidopsis and the family Brassicaceae. We promote the utility of such a "model clade" and make suggestions for building global networks to support future studies in the model order Brassicales.

A dataset of high-quality 7k SNP profiles of 1,038 unique Eurasian grapevine varieties was used to infer the most likely grapevine migration events, a spatial ancestry estimation, and a model about the origin of Eurasian grapevine germplasm. The comparison of putative gene flow scenarios from Caucasus throughout Europe aided to fit the more reliable spreading routes around the Mediterranean Basin. The same dataset was also used to assess the population genetic diversity, structure, and relatedness of Italian varieties. These data suggested a different history between Northern and Southern Italian grapevines, appearing clearly split into two different clusters. Interestingly, the Italian genotypes were shown to be distinguishable from all the other Eurasian populations for the first time. The same SNP panel was used to determine parental relationships, identifying the main parents of traditional Italian and closely related cultivars. The parentage network suggested that Italian germplasm largely originated from a few key parents distributed into several geographical areas of genetic influence, with more or less large overlaps. These key cultivars are 'Bombino bianco', 'Garganega'/'Grecanico', 'Mantonico bianco', 'Orsolina'/'Coccalona nera', 'Muscat à petits grains blanc', 'Malvasia odorosissima', 'Sangiovese', 'Sciaccarello', 'Visparola' and 'Vulpea'. The pedigree reconstruction by fullsib and second-degree relationships highlighted the pivotal role of some cultivars, such as 'Visparola', until now scarcely known. A hypothetic migration of this variety from Southern to Northern Italy along the Eastern side, as well as 'Sangiovese' migration from Southern to Central Italy along the Western side might be supposed. Moreover, 'Muscat à petits grains blanc', mainly through its offspring 'Malvasia odorosissima', furnished a consistent contribution to the development of many aromatic grapes grown in the Northern-Western part of the Italian Peninsula. These results represent the most complete study of grapevine Italian population genetics that has been carried out until now on the Italian germplasm.

Nitrate uptake by plants primarily relies on two gene families: Nitrate transporter 1/peptide transporter (NPF) and Nitrate transporter 2 (NRT2). Here, we extensively characterized the NPF and NRT2 families in the durum wheat genome, revealing 211 NPF and 20 NRT2 genes. The two families share many Cis Regulatory Elements (CREs) and Transcription Factor binding sites, highlighting a partially overlapping regulatory system and suggesting a coordinated response for nitrate transport and utilization. Analyzing RNA-seq data from 9 tissues and 20 cultivars, we explored expression profiles and co-expression relationships of both gene families. We observed a strong correlation between nucleotide variation and gene expression within the NRT2 gene family, implicating a shared selection mechanism operating on both coding and regulatory regions. Furthermore, NPF genes showed highly tissue-specific expression profiles, while NRT2s were mainly divided in two co-expression modules, one expressed in roots (NAR2/NRT3 dependent) and the other induced in anthers and/ovaries during maturation. Our evidences confirmed that the majority of these genes were retained after small-scale duplication events, suggesting a neo- or sub-functionalization of many NPFs and NRT2s. Altogether, these findings indicate that the expansion of these gene families in durum wheat could provide valuable genetic variability useful to identify NUE-related and candidate genes for future breeding programs in the context of low-impact and sustainable agriculture.

Climate change poses a major threat on agriculture, thus on food security. Drought stress, a factor in climate change, is a major problem for barley production, since it simultaneously affects morphological, physiological and biochemical traits. The present work was conducted to provide comprehensive information regarding barley genotypes response and adaptation to drought stress by using a high throughput phenotyping approach. Different barley genotypes were grown in a controlled environment greenhouse. Control plants were kept fully irrigated at 100% field capacity (FC), while the treated plants were stressed by reducing irrigation to 50% of FC. The effects of water deficit on barley genotypes development in terms of early detection of plant response to stress. Morpho-physiological parameters were recorded using Scanalyzer 3D High Throughput Phenotyping platform together with more conventional phenotyping methods to identify and select a set of putative drought tolerant genotypes. The results showed significant differences among genotypes in drought stress response based on digital and traditional indices. Among the selected tolerant genotypes, the best performer was a doubled haploid line derived by a cross Roho×Ardhaoui.

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Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), commonly called the red palm weevil (RPW), is one of the most severe palm pests of the tropics and Mediterranean countries. In these latter countries, this pest attacked mainly Phoenix canariensis, the most used ornamental palm for gardens and urban green spots. However, still is not understood if this preference for this host palm is determined by the attractiveness of its volatile organic compounds (VOCs) or to resistance/avoidance mechanisms present in the other palm species. In this study we investigate, in electro-physiological and behavioral experiment, the role of the P. canariensis VOCs in this host preference behavior versus other three palm species: Chamaerops humilis, Trachycarpus fortunei and Washingtonia filifera. In EAG recordings P. canariensis VOC extracts, elicited a stronger response on RPW antennae rather than solvent or C. humilis extract, while similar responses were recorded using W. filifera and T. fortunei extracts. In dual choice arena bioassays, females of RPW showed feeding preference for the P. canariensis in comparison with T. fortunei and C. humilis. Overall, the present findings suggest that this preference behavior is mediated by VOCs emitted.

The EuroVegChecklist (EVC) is currently considered to be the basis for the phytosociological classification of the European vegetation, and it is therefore important that it is continually updated, improved and refined. This article revises the nomenclature of three Mediterranean grassland-related classes and some of their contents down to the rank of association, namely Helianthemetea guttati, Poetea bulbosae and Stipo giganteae-Agrostietea castellanae, on the basis of the fourth edition of the International Code of Phytosociological Nomenclature (ICPN). The results show that the correct author citations for the first two classes are as follows: Tuberarietea guttatae Braun-Blanquet 1973, and Poetea bulbosae Rivas Goday et Rivas-Martínez ex Navarro Andrés et Valle Gutiérrez 1984. Instead, the name Stipo giganteae-Agrostietea castellanae is invalid and it is replaced by the new class name Agrostietea castellanae. In addition, we designate the neotype of the association Corynephoro articulati-Helianthemetum guttati (Corynephoro divaricati-Tuberarietum guttatae nom. corr. nov. et mut. nov.), the lectotype of the Poo bulbosae-Trifolietum subterranei, and mutate the names Helianthemetalia guttati and Helianthemion guttati to Tuberarietalia guttatae and Tuberarion guttatae, respectively, as well as two names of associations with a name-giving taxon belonging to the genus Tuberaria.

For ninety years, the alliance name Festucion valesiacae has been widely and almost exclusively used to designate the alliance of steppic, xeric grasslands on deep soils from Central Europe to western Ukraine. However, there is an earlier, hardly used heterotypic synonym, the Festucion sulcatae, that would be the correct name according to the rules [recte: Festucion rupicolae nom. corr.]. In order to preserve a well-established name, we propose to conserve the name Festucion valesiacae against the name Festucion sulcatae. In addition, we typify the name Festucion rupicolae Soó 1930 nom. corr. with the association Festuco rupicolae-Stipetum pennatae Soó 1930 nom. corr., for which we also select a neotype. This proposal is supported by the fact that the alliance Festucion valesiacae is the conserved type of the order Festucetalia valesiacae.

Plant-derived nanovesicles (PDNVs) have become attractive alternatives to mammalian cell-derived extracellular vesicles (EVs) both as therapeutic approaches and drug-delivery vehicles. In this study, we isolated tomato fruit-derived NVs and separated them by the iodixanol density gradient ultracentrifugation (DGUC) into twelve fractions. Three visible bands were observed at densities 1.064 ± 0.007 g/mL, 1.103 ± 0.006 g/mL and 1.122 ± 0.012 g/mL. Crude tomato PDNVs and DGUC fractions were characterized by particle size-distribution, concentration, lipid and protein contents as well as protein composition using mass spectrometry-based proteomics. Cytotoxicity and anti-inflammatory activity of the DGUC fractions associated to these bands were assessed in the lipopolysaccharide (LPS)-stimulated human monocytic THP-1 cell culture. The middle and the low-density visible DGUC fractions of tomato PDNVs showed a significant reduction in LPS-induced inflammatory IL-1? cytokine mRNA production. Functional analysis of proteins identified in these fractions reveals the presence of 14-3-3 proteins, endoplasmic reticulum luminal binding proteins and GTP binding proteins associated to gene ontology (GO) term GO:0050794 and the regulation of several cellular processes including inflammation. The most abundant middle-density DGUC fraction was loaded with curcumin using direct loading, sonication and extrusion methods and anti-inflammatory activity was compared. The highest entrapment efficiency and drug loading capacity was obtained by direct loading. Curcumin loaded by sonication increased the basal anti-inflammatory activity of tomato PDNVs.

In qualità di membro dalla commissione tecnico scientifica della regione Puglia sono stati presentati risultati di quanto fatto dalla suddetta commissione per la registrazione di leguminose e cereali pugliesi nel registro regionale delle varietà a rischio di estinzione.

The intensive application of agro-chemicals, and in particular of copper (Cu)-based compounds, causes increasing pollution of agricultural land, with serious risks for human health. Aromatic plants used for purposes other than food, can be considered for the exploitation and/or remediation of metal-polluted substrates, since contamination of the essential oils by these metals was not significant. Myrtle (Myrtus communis L.) is a Mediterranean evergreen shrub whose essential oil has many commercial applications. In this work, the effect of an excess of Cu in respect to control conditions was assessed on M. communis growth and foliar terpene composition. Metal accumulation in roots and shoots was also evaluated for the possible use of this species in phytoremediation. The amount of Cu applied in our experiments minimally affected the terpene profiles of in vitro grown plants, whereas no variations were detected in in vivo plants. The presence of the metal in the soil did not significantly impair plant growth, thus allowing its cultivation on polluted substrates. On the other hand, the amount of Cu in the plant was not enough to result in a significant reduction of Cu levels in the soil. Therefore, myrtle plants proved to be good candidates for the re-vegetation of Cu-contaminated lands.

Exposing fruits and vegetables to UVB radiation post-harvest is a technique used to modify secondary metabolites and prolong their shelf life. The aim of the present study was to evaluate the effects of UVB irradiation on the chemical and physical characteristics of fig cv. Dottato fruits. The UVB irradiation was 2.26 Wm-2. Two exposure times were carried out: 10 and 60 min resulting in a UVB dose of 1.4 and 8.1 kJm-2, respectively. In the control, the UVB was eliminated by a polyester film (control -UVB). After treatment, the fig fruits were stored and analyzed at different times until decay. Quality parameters (decay, weight loss, color, chlorophyll, and firmness) and physicochemical parameters (soluble solids content, pH parameters, and titratable acidity) were positively influenced by irradiation. Total and individual sugars increased gradually during the storage period in both the skin and the flesh, with glucose being higher after 10 days in the UVB treated samples. Total carotenoid content increased gradually during the storage period, with a marked increase in the +UVB fruit. The content of total and individual polyphenols was positively influenced by UVB treatment, with the UVB treated samples showing the highest values at both 7 and 10 days. The study showed an increase in by-products in both the skin and the flesh. This research confirms the effectiveness of UVB radiation in improving the nutritional qualities and shelf life of Ficus carica fruits.

Biostimulants (BSs) are natural materials (i.e., organic or inorganic compounds, and/or microorganisms) having beneficial effects on plant growth and productivity, and able to improve resilience/tolerance to biotic and abiotic stresses. Therefore, they represent an innovative alternative to the phyto- and agrochemicals, being environmentally friendly and a valuable tool to cope with extreme climate conditions. The objective of this study was to investigate the effects of several biomolecules (i.e., Xylanase, ?-Glucosidase, Chitinase, and Tramesan), alone or in combinations, on lettuce plant growth and quality. With this aim, the influence of these biomolecules on biomass, pigment content, and antioxidant properties in treated plants were investigated. Our results showed that Xylanase and, to a lesser extent, ?-Glucosidase, have potentially biostimulant activity for lettuce cultivation, positively influencing carotenoids, total polyphenols, and ascorbic acid contents; similar effects were found with respect to antioxidative properties. Furthermore, the effect of the more promising molecules (Xylanase and ?-Glucosidase) was also evaluated in kiwifruit cultured cells to test their putative role as sustainable input for plant cell biofactories. The absence of phytotoxic effects of both molecules at low doses (0.1 and 0.01 ?M), and the significantly enhanced cell biomass growth, indicates a positive impact on kiwifruit cells.

Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass production of hypocotyl explants for micrografting, as a continuous source for haploid and doubled haploid plants, and for germplasm conservation. It is so far the only pathway for the genetic modification of grapevines through transformation. The single-cell origin of somatic embryos makes them an ideal explant for mutation breeding as the resulting mutants will be chimera-free. In the present research, two combinations of plant growth regulators and different explants from flower buds at two stages of maturity were tested in regard to the efficiency of callusing and embryo formation from the callus produced in three white grape cultivars. Also, the treatment of somatic embryos with the chemical mutagen ethyl methanesulfonate (EMS) was optimised. Medium 2339 supplemented with ?-naphthoxyacetic acid (5 ?M) and 6-benzylaminopurine (BAP--9.0 ?M) produced significantly more calluses than medium 2337 supplemented with 2,4-dichlorophenoxyacetic acid (4.5 ?M) and BAP (8.9 ?M) in all explants. The calluses produced on medium 2337 were harder and more granular and produced more SEs. Although the stage of the maturity of floral bud did not have a significant effect on the callusing of the explants, calluses produced from immature floral bud explants in the premeiotic stage produced significantly more SEs than those from more mature floral buds. Overall, immature ovaries and cut floral buds exposing the cut ends of filaments, style, etc., tested for the first time in grapevine SE, produced the highest percentage of embryogenic calluses. It is much more efficient to cut the floral bud and culture than previously reported explants such as anthers, ovaries, stigmas and styles during the short flowering period when the immature flower buds are available. When the somatic embryos of the three cultivars were incubated for one hour with 0.1% EMS, their germination was reduced by 50%; an ideal treatment considered to obtain a high frequency of mutations for screening. Our research findings will facilitate more efficient SE induction in grapevines and inducing mutations for improving individual traits without altering the genetic background of the cultivar.

Climate change is favouring the northward shift of Mediterranean oaks which are expanding their ranges at their leading edges. However, Mediterranean oaks have recalcitrant seeds (desiccation sensitive) that do not form seed banks, depending on climate conditions after seed fall and interactions between genetic determinants to germinate. Here we investigate the potential adaptation of Quercus suber range-wide populations to increasing spring temperature in germination timing and rates. We sowed 701 acorns from 9 populations from 10 mother trees each, at 15, 20 and 25 °C and monitored germination daily during 90 days. We modelled germination timing through Cox's proportional-hazards models, assessed populations' adaptation to spring temperature transfer distances and quantified the effect of acorn mass and storage duration on germination probability and timing with fixed-effects models. We used the mixed-effects models to predict germination climatic niches under current and RCP 8.5 2080 scenarios. Differences in germination timing were due to both the population origin and temperature treatment; germination rates showed a sub-optimality towards warmer-than-origin temperatures. The timing of germination decreased along with spring temperatures increment, with germination in 2080 predicted to occur 12 days earlier than to date in central Iberia. Warmer spring temperatures significantly accelerate the germination of recalcitrant Mediterranean species, which could alter seedlings' developmental environment and ultimately populations' regeneration and species composition. As such, germination timing should receive more attention from scientists and stakeholders and should be included in forest vulnerability assessments and assisted migration programs aiming at long-term forest regenera- tion to adapt forests to climate change.

Irrigation is key to increasing crop yield and meeting the global demand for food. This study reports the assessment of tomato water consumption by bioristor, a new in vivo an Organic ElectroChemical Transistor-based biosensor. Bioristor enables direct, real-time acquisition of biophysical information about the plant's water requirements directly from the plant sap, and thus the water input can be adjusted accordingly. The aim of this study is to demonstrate the efficacy of bioristor in rapidly detecting changes in the plant's water status enhancing water use and irrigation efficiency in tomato cultivation with significant savings in the water supply. To this end, experiments were carried out in 2018 and 2020 in Parma (Italy) in tomato fields under different water regimes. The sensor response index (R) produced by bioristor recorded the real time plant health status, highlighting an excess in the water supplied as well as the occurrence of drought stress during the growing season. In both years, bioristor showed that the amount of water supplied could have been reduced by 36% or more. Bioristor also measured the timing and duration of leaf wetting: 438 h and 409 h in 2018 and 2020, respectively. These results open up new perspectives in irrigation efficiency and in more sustainable approaches to pesticide application procedures.

New mutations provide the raw material for evolution and adaptation. The distribution of fitness effects (DFE) describes the spectrum of effects of new mutations that can occur along a genome, and is, therefore, of vital interest in evolutionary biology. Recent work has uncovered striking similarities in the DFE between closely related species, prompting us to ask whether there is variation in the DFE among populations of the same species, or among species with different degrees of divergence, that is whether there is variation in the DFE at different levels of evolution. Using exome capture data from six tree species sampled across Europe we characterized the DFE for multiple species, and for each species, multiple populations, and investigated the factors potentially influencing the DFE, such as demography, population divergence, and genetic background. We find statistical support for the presence of variation in the DFE at the species level, even among relatively closely related species. However, we find very little difference at the population level, suggesting that differences in the DFE are primarily driven by deep features of species biology, and those evolutionarily recent events, such as demographic changes and local adaptation, have little impact.