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Grapevines store non-structural carbohydrates (NSC) during late summer to sustain plant development at the onset of the following spring’s growth. Starch is the main stored carbohydrate, found in the wood-ray parenchyma of roots and canes. Although the relationship between hydraulic and plant photosynthetic performance is well-recognized, little research has been done on the long-term effects of drought in grapevines adopting different strategies to cope with water stress (i.e. isohydric and anisohydric). We performed our study by exposing two different grape cultivars (Syrah and Cabernet Sauvignon) to a short but severe drought stress, at two stages of the growing season (July and September). No marked differences in the physiological and hydraulic responses of the two varieties were found, probably due to our experimental conditions. However, anatomical and biochemical characterization of overwintering canes pointed out several interesting outcomes. We found a significant and parallel increase of starch and medullar ray number in both cultivars exposed to early water stress. We hypothesize that stressed vines limited their carbon allocation to growth, while shifting it to starch accumulation, with a most evident effect in the period of intense photosynthetic activity. We also speculate that a different aptitude to osmotic adjustment may underlay variation in starch increase and the specific involvement of bark NSC in the two cultivars.

The effects of two deficit irrigation regimes (50% ETC and 20% ETC) on skin and seed phenolic compounds of the red Vitis vinifera L. cultivar 'Refošk' were studied during the veraison - maturation period. The study was undertaken for two successive seasons in the Karst area of North-East Italy, where rainfall is scarce and subterranean drainage limits surface water. To trace the effect of water deficit on both phenol biosynthesis and berry size reduction, phenols were extracted from lyophilised, cryomilled skins and seeds using organic solvents and evaluated in skin and seeds, per units of both dry and fresh weight. The anthocyanin concentration was higher in 20% ETC early after veraison, whereas at harvest the difference was not significant. A decreasing trend with some fluctuations in proanthocyanidin (PA) concentration in skins and seeds was found during maturation with both water regimes. 20% ETC resulted in a decrease in skin PA concentration and increase in seed PA concentration during ripening. As regards PA structural characteristics, there was a trend for higher galloylation and polymerisation of seeds and skins during ripening with the 20% ETC irrigation regime, whereas the effect on prodelphinidins in skins was inconsistent. In 2018, when most of the differences in vine water status between irrigation regimes occurred during veraison - maturation, the concentrations of PAs were affected more significantly than the mean degree of polymerisation and percentage of galloylation. In contrast, in 2019, when differences in vine water status occurred in the pre-veraison-maturation period, the structural characteristics of PAs were affected more significantly than concentrations, especially galloylation of skins and polymerisation of seeds. The results of the study showed that the biosynthesis of PAs, their polymerization and galloylation can be significantly modulated by water deficit regimes applied in the vineyard, in combination with the meteorological course of the season. [ABSTRACT FROM AUTHOR]

The need to minimise the impact of phytosanitary treatments for disease control boosted researchers to implement techniques with less environmental impact. The development of technologies using molecular mechanisms based on the modulation of metabolism by short dsRNA sequences appears promising. The intrinsic fragility of polynucleotides and the high cost of these techniques can be circumvented by nanocarriers that protect the bioactive molecule enabling high efficiency delivery to the leaf surface and extending its half-life. In this work, a specific protocol was developed aiming to assess the best methodological conditions for the synthesis of low-size chitosan nanoparticles (NPs) to be loaded with nucleotides. In particular, NPs have been functionalised with partially purified Green Fluorescent Protein dsRNAs (GFP dsRNA) and their size, surface charge and nucleotide retention capacity were analysed. Final NPs were also stained with FITC and sprayed on Nicotiana benthamiana leaves to assess, by confocal microscopy, both a distribution protocol and the fate of NPs up to 6 days after application. Finally, to confirm the ability of NPs to increase the efficacy of dsRNA interference, specific tests were performed: by means of GFP dsRNA-functionalised NPs, the nucleotide permanence during time was assessed both in vitro on detached wild-type N. benthamiana leaves and in planta; lastly, the inhibition of Botrytis cinerea on single leaves was also evaluated, using a specific fungal sequence (Bc dsRNA) as the NPs' functionalising agent. The encouraging results obtained are promising in the perspective of long-lasting application of innovative treatments based on gene silencing. [ABSTRACT FROM AUTHOR]

Xylem embolism is one of the possible outcomes of decreasing xylem pressure when plants face drought. Recent studies have proposed a role for non-structural carbohydrates (NSCs) in osmotic pressure generation, required for refilling embolized conduits. Potted cuttings of grapevine Grenache and Barbera, selected for their adaptation to different climatic conditions, were subjected to a drought stress followed by re-irrigation. Stem embolism rate and its recovery were monitored in vivo by X-ray micro-computed tomography (micro-CT). The same plants were further analyzed for xylem conduit dimension and NSC content. Both cultivars significantly decreased Ψpd in response to drought and recovered from xylem embolism after re-irrigation. However, although the mean vessel diameter was similar between the cultivars, Barbera was more prone to embolism. Surprisingly, vessel diameter was apparently reduced during recovery in this cultivar. Hydraulic recovery was linked to sugar content in both cultivars, showing a positive relationship between soluble NSCs and the degree of xylem embolism. However, when starch and sucrose concentrations were considered separately, the relationships showed cultivar-specific and contrasting trends. We showed that the two cultivars adopted different NSC-use strategies in response to drought, suggesting two possible scenarios driving conduit refilling. In Grenache, sucrose accumulation seems to be directly linked to embolism formation and possibly sustains refilling. In Barbera, maltose/maltodextrins could be involved in a conduit recovery strategy via the formation of cell-wall hydrogels, likely responsible for the reduction of conduit lumen detected by micro-CT. [ABSTRACT FROM AUTHOR]

BACKGROUND: In recent years, biofungicides have drawn increasing interest in vineyards for a more sustainable integrated and copper‐limited pest management. Among alternatives, botanicals could represent valuable tools, being rich sources of biologically active compounds. Conversely to the well‐known antioxidant and biological properties in relation to health benefits, investigation on bioactivity of hot pungent Capsicum sp. products against fungal phytopathogens in vineyards is still scarce. Therefore, the present study aimed at exploring the biologically active compounds profile of a chili pepper (Capsicum chinense Jacq.) pod extract and its antimicrobial properties against some of the major fungal and Oomycetes pathogens of grapevine, including Botrytis cinerea Pers., Guignardia bidwellii (Ellis) Viala & Ravaz and Plasmopara viticola (Berk. & M.A. Curtis) Berl. & De Toni. RESULTS: The ethyl acetate‐extracted oleoresin from the most pungent varieties was rich in capsaicinoids and polyphenols (371.09 and 268.5 μg mg−1 dry weight, respectively). Capsaicin and dihydrocapsaicin, hydroxycinnamic and hydroxybenzoic acids and quercetin derivatives were the most abundant, while carotenoids represented only a minor fraction. The oleoresin was efficient to inhibit all three pathogenic fungi and ED50 values were determined, evidencing that G. bidwellii was the more sensitive (0.233 ± 0.034 mg mL−1). CONCLUSION: The results suggested a potentiality of chili pepper extract for the control of some important grapevine pathogens, their possible application being helpful for the recommended limitation in extensive use of copper in vineyard. The complex mixture of high amounts of capsaicinoids, associated to specific phenolic acids and other minor bioactive components might contribute to the observed antimicrobial action of chili pepper extract. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Premise: Despite great attention given to the relationship between plant growth and carbon balance in alpine tree species, little is known about shrubs at the treeline. We hypothesized that the pattern of main nonstructural carbohydrates (NSCs) across elevations depends on the interplay between phenotypic trait plasticity, plant–plant interaction, and elevation. Methods: We studied the pattern of NSCs (i.e., glucose, fructose, sucrose, and starch) in alpine stands of Vaccinium myrtillus (above treeline) across an elevational gradient. In the same plots, we measured key growth traits (i.e., anatomical stem features) and shrub cover, evaluating putative relationships with NSCs. Results: Glucose content was positively related with altitude, but negatively related with shrub cover. Sucrose decreased at high altitude and in older populations and increased with higher percentage of vascular tissue. Starch content increased at middle and high elevations and in stands with high shrub cover. Moreover, starch content was negatively related with the number of xylem rings and the percentage of phloem tissue, but positively correlated with the percentage of xylem tissue. Conclusions: We found that the increase in carbon reserves across elevations was uncoupled from plant growth, supporting the growth limitation hypothesis, which postulates NSCs accumulate at high elevation as a consequence of low temperature. Moreover, the response of NSC content to the environmental stress caused by elevation was buffered by phenotypic plasticity of plant traits, suggesting that, under climate warming conditions, shrub expansion due to enhanced plant growth would be pronounced in old but sparse stands. [ABSTRACT FROM AUTHOR]

In striking analogy with Saccharomyces cerevisiae , etiolated pea stem mitochondria did not show appreciable Ca2+ uptake. Only treatment with the ionophore ETH129 (which allows electrophoretic Ca2+ equilibration) caused Ca2+ uptake followed by increased inner membrane permeability, membrane depolarization and Ca2+ release. Like the permeability transition (PT) of mammals, yeast and Drosophila, the PT of pea stem mitochondria was stimulated by diamide and phenylarsine oxide and inhibited by Mg-ADP and Mg-ATP, suggesting a common underlying mechanism; yet, the plant PT also displayed distinctive features: (i) as in mammals it was desensitized by cyclosporin A, which does not affect the PT of yeast and Drosophila; (ii) similarly to S. cerevisiae and Drosophila it was inhibited by Pi, which stimulates the PT of mammals; (iii) like in mammals and Drosophila it was sensitized by benzodiazepine 423, which is ineffective in S. cerevisiae ; (iv) like what observed in Drosophila it did not mediate swelling and cytochrome c release, which is instead seen in mammals and S. cerevisiae. We find that cyclophilin D, the mitochondrial receptor for cyclosporin A, is present in pea stem mitochondria. These results indicate that the plant PT has unique features and suggest that, as in Drosophila, it may provide pea stem mitochondria with a Ca2+ release channel. [ABSTRACT FROM AUTHOR]

Background: Senescence is a key developmental process occurring during the life cycle of plants that can be induced also by environmental conditions, such as starvation and/or darkness. During senescence, strict control of genes regulates ordered degradation and dismantling events, the most remarkable of which are genetically programmed cell death (PCD) and, in most cases, an upregulation of flavonoid biosynthesis in the presence of light. Flavonoids are secondary metabolites that play multiple essential roles in development, reproduction and defence of plants, partly due to their well-known antioxidant properties, which could affect also the same cell death machinery. To understand further the effect of endogenously-produced flavonoids and their interplay with different environment (light or dark) conditions, two portions (red and green) of a senescing grapevine callus were used to obtain suspension cell cultures. Red Suspension cell Cultures (RSC) and Green Suspension cell Cultures (GSC) were finally grown under either dark or light conditions for 6 days. Results: Darkness enhanced cell death (mainly necrosis) in suspension cell culture, when compared to those grown under light condition. Furthermore, RSC with high flavonoid content showed a higher viability compared to GSC and were more protected toward PCD, in accordance to their high content in flavonoids, which might quench ROS, thus limiting the relative signalling cascade. Conversely, PCD was mainly occurring in GSC and further increased by light, as it was shown by cytochrome c release and TUNEL assays. Conclusions: Endogenous flavonoids were shown to be good candidates for exploiting an efficient protection against oxidative stress and PCD induction. Light seemed to be an important environmental factor able to induce PCD, especially in GSC, which lacking of flavonoids were not capable of preventing oxidative damage and signalling leading to senescence. [ABSTRACT FROM AUTHOR]

In the present study, an antibody raised against a peptide sequence of rat bilitranslocase (anti-peptide Ab) was tested on microsomal proteins obtained from red grape berry skin. Previously, this antibody had demonstrated to recognize plant membrane proteins associated with flavonoid binding and transport. Immuno-proteomic assays identified a number of proteins reacting with this particular antibody, suggesting that the flavonoid binding and interaction may be extended not only to carriers of these molecules, but also to enzymes with very different functions. One of these proteins is a pathogenesis-related (PR) class IV chitinase, whose in vitro chitinolytic activity was modulated by two of the most representative flavonoids of grape, quercetin and catechin, as assessed by both spectrophotometric and fluorimetric assays in grape microsomes and commercial enzyme preparations. The effect of these flavonoids on the catalysis and its kinetic parameters was also evaluated, evidencing that they determine a hormetic dose-dependent response. These results highlight the importance of flavonoids not only as antioxidants or antimicrobial effectors, but also as modulators of plant growth and stress response. Implications of the present suggestion are here discussed in the light of environment and pesticide-reduction concerns. [ABSTRACT FROM AUTHOR]

The synthesis of ATP in mitochondria is dependent on a low permeability of the inner membrane. Nevertheless, mitochondria can undergo an increased permeability to solutes, named permeability transition (PT) that is mediated by a permeability transition pore (PTP). PTP opening requires matrix Ca2+ and leads to mitochondrial swelling and release of intramembrane space proteins (e.g., cytochrome c). This feature has been initially observed in mammalian mitochondria and tentatively attributed to some components present either in the outer or inner membrane. Recent works on mammalian mitochondria point to mitochondrial ATP synthase dimers as physical basis for PT, a finding that has been substantiated in yeast and Drosophila mitochondria. In plant mitochondria, swelling and release of proteins have been linked to programmed cell death, but in isolated mitochondria PT has been observed in only a few cases and in plant cell cultures only indirect evidence is available. The possibility that mitochondrial ATP synthase dimers could function as PTP also in plants is discussed here on the basis of the current evidence. Finally, a hypothetical explanation for the origin of PTP is provided in the framework of molecular exaptation. [ABSTRACT FROM AUTHOR]

Flavonoids represent one of the most important molecules of plant secondary metabolism, playing many different biochemical and physiological roles. Although their essential role in plant life and human health has been elucidated by many studies, their subcellular transport and accumulation in plant tissues remains unclear. This is due to the absence of a convenient and simple method to monitor their transport. In the present work, we suggest an assay able to follow in vivo transport of quercetin, the most abundant flavonoid in plant tissues. This uptake was monitored using 2-aminoethoxydiphenyl borate (DPBA), a fluorescent probe, in non-pigmented Vitis vinifera cell cultures. [ABSTRACT FROM AUTHOR]

The article focuses on a study related to the synthesis of flavonoids in grapevine. The study highlights the role of flavonoids in biosynthesis, transport and stress response of plant cell. The study further mentions that a multidrug and toxic compound extrusion (MATE) and ATP-binding cassette (ABC) transporters have been demonstrated in tonoplast of grape berry.

Programmed cell death (PCD) is a finely tuned process of multicellular organisms. In higher plants, PCD regulates many developmental processes and the response of host plants to incompatible pathogens (hypersensitive response). Four types of PCD have been described in plants, mainly associated to vacuole rupture, that is followed by the appearance of the typical PCD hallmarks (i.e. nuclear DNA fragmentation and cell shrinkage). However, in some cases vacuole collapse is preceded by an early alteration of other subcellular organelles, such as mitochondria. In particular, the central role played by mitochondria in PCD has been largely recognised in animal cells. This review deals with the involvement of mitochondria in the manifestation of plant PCD, in comparison to that described in animal PCD. The main hallmark, connecting animal and plant PCD via mitochondria, is represented by the release of cytochrome c and possibly other chemicals such as nucleases, which may be accomplished by different mechanisms, involving both swelling and non-swelling of the organelles. [ABSTRACT FROM AUTHOR]

Pea stem mitochondria, resuspended in a KCl medium (de-energized mitochondria), underwent a swelling, as a consequence of K+ entry, that was inhibited by ATP. This inhibition was partially restored by GTP and diazoxide (K+ ATP channel openers). In addition, glyburide and 5-hydroxydecanate (K+ ATP channel blockers) induced an inhibition of the GTP-stimulated swelling. Mitochondrial swelling was inhibited by H2O2, but stimulated by NO. The same type of responses was also obtained in succinate-energized mitochondria. When the succinate-dependent transmembrane electrical potential (ΔΨ) had reached a steady state, the addition of KCl induced a dissipation that was inhibited by H2O2 and stimulated by NO. The latter stimulation was prevented by carboxy-PTIO, a NO scavenger. Phenylarsine oxide (a thiol oxidant) and NEM (a thiol blocker) stimulated the KCl-induced dissipation of ΔΨ, while DTE prevented this effect in both cases. In addition, DTE transiently inhibited the NO-induced dissipation of ΔΨ, but then it caused a more rapid collapse. These results, therefore, show that the plant mitochondrial K+ ATP channel resembles that present in mammalian mitochondria and that it appears to be modulated by dithiol–disulfide interconversion, NO and H2O2. The aperture of this channel was linked to the partial rupture of the outer membrane. The latter effect led to a release of cytochrome c, thus suggesting that this release may be involved in the manifestation of programmed cell death. [ABSTRACT FROM AUTHOR]

Studying changes in partitioning of dry matter and nonstructural carbohydrates (NSC) content in both aboveground and underground perennial tissues in drought-affected grapevines could provide insights into plant response and carbon allocation strategies during stress periods. The analysis of soluble NSC and starch content in leaf petioles, due to their role in hydraulic segmentation, should also be considered. In the present research, these aspects have been investigated in Merlot grapevines grown in pots and subjected to progressive and increasing soil dehydration, and in well-irrigated vines. Drought conditions caused drastic reduction of shoot elongation and total plant leaf area development in favor of a greater biomass allocation and partitioning towards roots, where most of the NSC reserves were also conserved. Dry matter content of the perennial organs increased in stressed vines due to growth reduction, allocation of carbon reserves and possible anatomical modifications. Vines subjected to drought showed a higher NSC content in petioles, supporting the hypothesis that they are involved as compatible solutes in osmotic adjustments. [ABSTRACT FROM AUTHOR]

The electrical potential difference (δΨ) generation and ATP synthesis due to α-keto acid-stimulated alternative oxidase activity in pea leaf mitochondria, energized by malate plus glutamate, was studied. In the absence of α-keto acids, δΨwas ca. 60% collapsed by KCN or antimycin A (AA), while the remaining part (ca. 30%) was dissipated by salicyl-hydroxamic acid (SHAM). The presence of α-keto acids (pyruvate or iodoacetate), known to stimulate the alternative oxidase, caused a decrease in the KCN- or AA-sensitive component of δΨ(ca. 30%), which was accompanied by an increase in the SHAM-sensitive one. When mitochondria were energized by exogenous NADH, succinate or duroquinol, δΨ was completely collapsed by KCN or AA, either in the presence or absence of α-keto acids. ADP partially collapsed the SHAM-sensitive component of δΨ, and oligomycin restored this dissipation. In the presence of pyruvate or iodoacetate, ATP content in KCN-treated mitochondria was ca. 40% of control, while that in SHAM plus KCN-inhibited mitochondria was negligible. ADP stimulated oxygen uptake only in the presence of KCN (respiratory control ratio = 1.7), but not in the presence of KCN plus SHAM. It is concluded that α-keto acids, stimulating the oxidation of NAD-linked substrates through the alternative oxidase, lead to an increase in the SHAM-sensitive component of δΨvia complex I, thus increasing the contribution of this pathway to ATP synthesis when the cyto-chrome oxidase is restricted. [ABSTRACT FROM AUTHOR]

In grapevine, the anatomy of xylem conduits and the non-structural carbohydrates (NSCs) content of the associated living parenchyma are expected to influence water transport under water limitation. In fact, both NSC and xylem features play a role in plant recovery from drought stress. We evaluated these traits in petioles of Cabernet Sauvignon (CS) and Syrah (SY) cultivars during water stress (WS) and recovery. In CS, the stress response was associated to NSC consumption, supporting the hypothesis that starch mobilization is related to an increased supply of maltose and sucrose, putatively involved in drought stress responses at the xylem level. In contrast, in SY, the WS-induced increase in the latter soluble NSCs was maintained even 2 days after re-watering, suggesting a different pattern of utilization of NSC resources. Interestingly, the anatomical analysis revealed that conduits are constitutively wider in SY in well-watered (WW) plants, and that water stress led to the production of narrower conduits only in this cultivar. [ABSTRACT FROM AUTHOR]

The xylem is a complex system that includes a network of dead conduits ensuring long-distance water transport in plants. Under ongoing climate changes, xylem embolism is a major and recurrent cause of drought-induced tree mortality. Non-structural carbohydrates (NSC) play key roles in plant responses to drought and frost stress, and several studies putatively suggest their involvement in the regulation of xylem water transport. However, a clear picture on the roles of NSCs in plant hydraulics has not been drawn to date. We summarize the current knowledge on the involvement of NSCs during embolism formation and subsequent hydraulic recovery. Under drought, sugars are generally accumulated in xylem parenchyma and in xylem sap. At drought-relief, xylem functionality is putatively restored in an osmotically driven process involving wood parenchyma, xylem sap and phloem compartments. By analyzing the published data on stem hydraulics and NSC contents under drought/frost stress and subsequent stress relief, we found that embolism build-up positively correlated to stem NSC depletion, and that the magnitude of post-stress hydraulic recovery positively correlated to consumption of soluble sugars. These findings suggest a close relationship between hydraulics and carbohydrate dynamics. We call for more experiments on hydraulic and NSC dynamics in controlled and field conditions. [ABSTRACT FROM AUTHOR]

The reduction of synthetic chemistry use in modern viticulture relies on either the biological control of microorganisms or the induction of pathogenesis-related proteins. In the present study, the effects of hydro-alcoholic plant extracts (PEs) (i.e., by-products of Vitis vinifera L., leaves of Olea europaea L. and Ailanthus altissima (Mill.) Swingle) were tested on purified enzymes activity involved in plant-pathogen interactions. The polyphenolic composition was assayed and analyzed to characterize the extract profiles. In addition, suspension cell cultures of grapevine were treated with PEs to study their modulation of chitinase activity. Application of grape marc's PE enhanced chitinase activity at 4 g L−1. Additionally, foliar treatment of grape marc's PE at two doses (4 g L−1 and 800 g L−1) on grapevine cuttings induced a concentration-dependent stimulation of chitinase activity. The obtained results showed that the application of bioactive compounds based on PEs, rich in phenolic compounds, was effective both at in vitro and ex/in vivo level. The overall effects of PEs on plant-pathogen interaction were further discussed by applying a multi-criteria decision analysis, showing that grape marc was the most effective extract. [ABSTRACT FROM AUTHOR]

The Nutrient Use Efficiency in intensive agriculture is lower than 50% for macronutrients. This feature results in unsustainable financial and environmental costs. Nanofertilizers are a promising application of nanotechnology in agriculture. The use of nanofertilizers in an efficient and safe manner calls for knowledge about the actual effects of nanoproducts on the plant metabolism and eventually on the carrier release kinetics and nutrient accumulation. Hydroxyapatite (Ca10(PO4)6(OH)2) nanoparticles (nHA) have an interesting potential to be used as nanofertilizers. In this study, the effects of different nHA solutions stabilized with carboxymethylcellulose (CMC) were evaluated on germination, seedling growth, and metabolism of Solanum lycopersicum L., used as model species. Our observations showed that the percentage germination of S. lycopersicum is not influenced by increasing concentrations of nHa, while root elongation is strongly stimulated. Tomato plants grown in hydroponics in the presence of nHA have not suffered phytotoxic effects. We conclude that nHA had nontoxic effects on our model plant and therefore it could be used both as a P supplier and carrier of other elements and molecules. [ABSTRACT FROM AUTHOR]