Monooxygenases and Antibiotic Resistance: A Focus on Carbapenems.

Carbapenems are a group of broad-spectrum beta-lactam antibiotics that in many cases are the last effective defense against infections caused by multidrug-resistant bacteria, such as some strains of Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii. Resistance to carbapenems has emerged and is beginning to spread, becoming an ongoing public-health problem of global dimensions, causing serious outbreaks, and dramatically limiting treatment options. This paper reviews the role of flavin monooxygenases in antibiotic resistance, with a specific focus on carbapenem resistance and the recently discovered mechanism mediated by Baeyer-Villiger monooxygenases. Flavin monooxygenases are enzymes involved in the metabolism and detoxification of compounds, including antibiotics. Understanding their role in antibiotic resistance is crucial. Carbapenems are powerful antibiotics used to treat severe infections caused by multidrug-resistant bacteria. However, the rise of carbapenem-resistant strains poses a significant challenge. This paper explores the mechanisms by which flavin monooxygenases confer resistance to carbapenems, examining molecular pathways and genetic factors. Additionally, this paper highlights the discovery of Baeyer-Villiger monooxygenases' involvement in antibiotic resistance. These enzymes catalyze the insertion of oxygen atoms into specific chemical bonds. Recent studies have revealed their unexpected role in promoting carbapenem resistance. Through a comprehensive analysis of the literature, this paper contributes to the understanding of the interplay between flavin monooxygenases, carbapenem resistance, and Baeyer-Villiger monooxygenases. By exploring these mechanisms, it aims to inform the development of strategies to combat antibiotic resistance, a critical global health concern.

Role of Cytochrome P450 Enzyme in Plant Microorganisms' Communication: A Focus on Grapevine.

Cytochromes P450 are ancient enzymes diffused in organisms belonging to all kingdoms of life, including viruses, with the largest number of P450 genes found in plants. The functional characterization of cytochromes P450 has been extensively investigated in mammals, where these enzymes are involved in the metabolism of drugs and in the detoxification of pollutants and toxic chemicals. The aim of this work is to present an overview of the often disregarded role of the cytochrome P450 enzymes in mediating the interaction between plants and microorganisms. Quite recently, several research groups have started to investigate the role of P450 enzymes in the interactions between plants and (micro)organisms, focusing on the holobiont Vitis vinifera. Grapevines live in close association with large numbers of microorganisms and interact with each other, regulating several vine physiological functions, from biotic and abiotic stress tolerance to fruit quality at harvest.

Juxtaposition of the Source-to-Sink Ratio and Fruit Exposure to Solar Radiation on cv. Merlot (Vitis vinifera L.) Berry Phenolics in a Cool versus Warm Growing Region.

The grapevine source-to-sink ratio and berry exposure to solar radiation both influence grape flavonoid biosynthesis and accumulation. Here, we compared these concepts on cv. Merlot in two different growing locations (Michigan (MI) and Friuli-Venezia Giulia (FVG), IT) to understand whether the environment influences flavonoid sensitivity to these parameters. Three levels of leaf removal (LR0, LR5, LR8) were implemented at the pea-size phenological stage to compare conditions of increased cluster light exposure with a decreasing vine source-to-sink ratio on berry flavonoid accumulation. Treatments did not affect total soluble solids (TSSs) or pH, but titratable acidity (TA) was lower in LR8 at harvest in both locations. LR5 increased anthocyanins and flavonols in MI but decreased most phenolics in FVG. The decreased expression of VviLAR1 and VviF3'5'Hh during ripening supported the lower concentrations of flavan-3-ol monomers and anthocyanins in FVG. In summary, flavonoid biosynthesis and accumulation were more sensitive to solar radiation than the source-to-sink ratio, and the vineyard environment dictated whether solar radiation was beneficial or detrimental to flavonoid biosynthesis.

Influence of freezing and heating conditions on grape seed flavan-3-ol extractability, oxidation, and galloylation pattern.

In cool-climate viticulture, the short growing season can influence grape seed maturation by reducing the apparent oxidation of flavan-3-ol monomers and associated increase in seed browning. A reduction in seed maturation increases the potential extraction of flavan-3-ol monomers into wine during maceration operations, heightening bitterness. Here, we carried out a 2 × 2 factorial experiment to test the ability of freezing and heating treatments to advance maturation (decrease flavan-3-ol, improve browning) of (Vitis vinifera L.) Pinot noir and Cabernet Sauvignon seeds over a 24-h incubation period. Only freezing significantly increased seed browning in both cultivars. Subsequent correlations with seed flavan-3-ol monomer concentrations suggest that freezing enhanced the oxidation of these compounds. Interestingly, natural ripening and freezing reduced galloylated flavan-3-ol monomers to a greater extent than non-galloylated ones. This study provides new information regarding the susceptibility of flavan-3-ol monomers to freezing and heating, and also suggests that freezing can advance the maturation the seeds of under-ripe red vinifera grapes.

Traditional in vitro strategies for sustainable production of bioactive compounds and manipulation of metabolomic profile in medicinal, aromatic and ornamental plants.

MAIN CONCLUSION: Precursor feeding, elicitation and culture medium parameters are traditional in vitro strategies to enhance bioactive compounds of medicinal, aromatic, and ornamental plants (MAOPs). Machine learning can help researchers find the best combination of these strategies to increase the secondary metabolites content of MAOPs. Many requirements for human life, from food, pharmaceuticals and cosmetics to clothes, fuel and building materials depend on plant-derived natural products. Essential oils, methanolic and ethanolic extracts of in vitro undifferentiated callus and organogenic cultures of medicinal, aromatic, and ornamental plants (MAOPs) contain bioactive compounds that have several applications for various industries, including food and pharmaceutical. In vitro culture systems provide opportunities to manipulate the metabolomic profile of MAOPs. Precursors feeding, elicitation and culture media optimization are the traditional strategies to enhance in vitro accumulation of favorable bioactive compounds. The stimulation of plant defense mechanisms through biotic and abiotic elicitors is a simple way to increase the production of secondary metabolites in different in vitro culture systems. Different elicitors have been applied to stimulate defense machinery and change the metabolomic profile of MAOPs in in vitro cultures. Plant growth regulators (PGRs), stress hormones, chitosan, microbial extracts and physical stresses are the most applied elicitors in this regard. Many other chemical tolerance-enhancer additives, such as melatonin and proline, have been applied along with stress response-inducing elicitors. The use of stress-inducing materials such as PEG and NaCl activates stress tolerance elicitors with the potential of increasing secondary metabolites content of MAOPs. The present study reviewed the state-of-the-art traditional in vitro strategies to manipulate bioactive compounds of MAOPs. The objective is to provide insights to researchers involved in in vitro production of plant-derived natural compounds. The present review provided a wide range of traditional strategies to increase the accumulation of valuable bioactive compounds of MAOPs in different in vitro systems. Traditional strategies are faster, simpler, and cost-effective than other biotechnology-based breeding methods such as genetic transformation, genome editing, metabolic pathways engineering, and synthetic biology. The integrate application of precursors and elicitors along with culture media optimization and the interpretation of their interactions through machine learning algorithms could provide an excellent opportunity for large-scale in vitro production of pharmaceutical bioactive compounds.

Modeling Agrobacterium-Mediated Gene Transformation of Tobacco (Nicotiana tabacum)-A Model Plant for Gene Transformation Studies.

The multilayer perceptron (MLP) topology of an artificial neural network (ANN) was applied to create two predictor models in Agrobacterium-mediated gene transformation of tobacco. Agrobacterium-mediated transformation parameters, including Agrobacterium strain, Agrobacterium cell density, acetosyringone concentration, and inoculation duration, were assigned as inputs for ANN-MLP, and their effects on the percentage of putative and PCR-verified transgenic plants were investigated. The best ANN models for predicting the percentage of putative and PCR-verified transgenic plants were selected based on basic network quality statistics. Ex-post error calculations of the relative approximation error (RAE), the mean absolute error (MAE), the root mean square error (RMS), and the mean absolute percentage error (MAPE) demonstrated the prediction quality of the developed models when compared to stepwise multiple regression. Moreover, significant correlations between the ANN-predicted and the actual values of the percentage of putative transgenes (R 2 = 0.956) and the percentage of PCR-verified transgenic plants (R 2 = 0.671) indicate the superiority of the established ANN models over the classical stepwise multiple regression in predicting the percentage of putative (R 2 = 0.313) and PCR-verified (R 2= 0.213) transgenic plants. The best combination of the multiple inputs analyzed in this investigation, to achieve maximum actual and predicted transgenic plants, was at OD 600 = 0.8 for the LB4404 strain of Agrobacterium × 300 µmol/L acetosyringone × 20 min immersion time. According to the sensitivity analysis of ANN models, the Agrobacterium strain was the most important influential parameter in Agrobacterium-mediated transformation of tobacco. The prediction efficiency of the developed model was confirmed by the data series of Agrobacterium-mediated transformation of an important medicinal plant with low transformation efficiency. The results of this study are pivotal to model and predict the transformation of other important Agrobacterium-recalcitrant plant genotypes and to increase the transformation efficiency by identifying critical parameters. This approach can substantially reduce the time and cost required to optimize multi-factorial Agrobacterium-mediated transformation strategies.

Vitis vinifera Production in Michigan: Factors and Trends Driving Cultivation Patterns.

Vinifera cultivation is a thriving and growing industry across the state of Michigan (MI), United States. Extensive time, funds, and effort have been applied by the industry to promote growth and the onset of new producers. Specifically, Vitis vinifera wine grapes, which have been cultivated in MI since the 1970s, have seen a rapid expansion and investment from both first-time and legacy growers. However, historically, the climate of MI presented a challenge for cultivation because of low growing season temperatures (GSTs), short growing seasons, and excessive precipitation at the time of harvest. Over time, two key factors have led the MI wine industry to overcome the challenging climate. First, as seen in the literature, there are noted changes in climate, especially since the late 1980s, leading to more favorable conditions for cultivation. Second, MI growers traditionally focused on V. vinifera cultivation, which is susceptible to low winter temperatures, selected less vulnerable regions within the state while also focusing on vine protection techniques. Given the rapid growth of the wine industry across MI, there is a need to understand suitability and its drivers to help all growers make economically impactful decisions on production and expansion of wine grapes. This article looked to study the suitability of MI vinifera across the state in two ways. Initially, through an extensive literature review, the key drivers and commonly noted trends guiding vinifera production were chronicled. Second, through a trend analysis of the key drivers of suitability, the study investigated how such variables are changing significantly over space and time. The results of this study expand the knowledge of cool climate agriculture production and suitability for cultivation and highlight the complexity of relating suitability drivers for non-cool climate to cool climate vinifera cultivation.

Betaine Aldehyde Dehydrogenase (BADH) vs. Flavodoxin (Fld): Two Important Genes for Enhancing Plants Stress Tolerance and Productivity.

Abiotic stresses, mainly salinity and drought, are the most important environmental threats that constrain worldwide food security by hampering plant growth and productivity. Plants cope with the adverse effects of these stresses by implementing a series of morpho-physio-biochemical adaptation mechanisms. Accumulating effective osmo-protectants, such as proline and glycine betaine (GB), is one of the important plant stress tolerance strategies. These osmolytes can trigger plant stress tolerance mechanisms, which include stress signal transduction, activating resistance genes, increasing levels of enzymatic and non-enzymatic antioxidants, protecting cell osmotic pressure, enhancing cell membrane integrity, as well as protecting their photosynthetic apparatus, especially the photosystem II (PSII) complex. Genetic engineering, as one of the most important plant biotechnology methods, helps to expedite the development of stress-tolerant plants by introducing the key tolerance genes involved in the biosynthetic pathways of osmolytes into plants. Betaine aldehyde dehydrogenase (BADH) is one of the important genes involved in the biosynthetic pathway of GB, and its introduction has led to an increased tolerance to a variety of abiotic stresses in different plant species. Replacing down-regulated ferredoxin at the acceptor side of photosystem I (PSI) with its isofunctional counterpart electron carrier (flavodoxin) is another applicable strategy to strengthen the photosynthetic apparatus of plants under stressful conditions. Heterologous expression of microbially-sourced flavodoxin (Fld) in higher plants compensates for the deficiency of ferredoxin expression and enhances their stress tolerance. BADH and Fld are multifunctional transgenes that increase the stress tolerance of different plant species and maintain their production under stressful situations by protecting and enhancing their photosynthetic apparatus. In addition to increasing stress tolerance, both BADH and Fld genes can improve the productivity, symbiotic performance, and longevity of plants. Because of the multigenic and complex nature of abiotic stresses, the concomitant delivery of BADH and Fld transgenes can lead to more satisfying results in desired plants, as these two genes enhance plant stress tolerance through different mechanisms, and their cumulative effect can be much more beneficial than their individual ones. The importance of BADH and Fld genes in enhancing plant productivity under stress conditions has been discussed in detail in the present review.

The Decrease of Leaf Dark Respiration during Water Stress Is Related to Leaf Non-Structural Carbohydrate Pool in Vitis vinifera L.

Dark respiration (Rd) is a fundamental plant process used to gain biomass and maintain plant physiological activity. It accounts for the metabolization of a large share of the carbon fixed by photosynthesis. However, Rd during conditions of severe plant water stress is still poorly understood. The decrease in leaf transpiration increases temperature, one of the most important drivers of leaf Rd. On the other hand, water stress decreases the pool of leaf carbohydrates, which are the most important substrate for respiration. The aim of the present work was to determine the impact of water shortage on leaf Rd in grapevine and understand the driving factors in modulating leaf Rd response under plant water stress conditions. Water stressed vines had lower Rd as the water shortage severity increased. Rd was correlated with leaf temperature in well-watered vines. Instead, in water stressed vines, Rd correlated with leaf soluble sugars. The decrease of leaf Rd in water stressed vines was due to the decrease of leaf non-structural carbohydrate that, under water stress conditions, exerted a limiting effect on Rd.

Canopy architecture and fruit microclimate, not ripening-related phytohormones, control phenylpropanoid accumulation in response to early leaf removal in 'Merlot' (Vitis vinifera L.) grapevines.

Early leaf removal (ELR) applied in the grapevine cluster zone at bloom or pre-bloom (PB) is a vineyard practice commonly utilized to reduce fruit disease and yield. In addition, the literature reports that ELR enhances fruit quality, however, little research has deciphered the potential factors regulating this response. In this work, the objective was to understand whether the increase in fruit quality in response to manual or mechanical leaf removal is due to changes in fruit-zone microclimate, vine physiology, or ripening/stress related hormone biosynthesis. In 'Merlot' (Vitis vinifera L.) vines, 60% of leaf area was removed from shoots in three ways: 1) manual removal of 5 leaves (PB-MA), 2) mechanical removal (PB-ME), and 3) simulated mechanical removal (PB-SIM), which was implemented by removing the distal portion of leaves on the first eight nodes to understand whether PB-ME improves fruit quality via enhanced microclimate conditions or plant stress. Yield was reduced in PB-ME and PB-SIM, while total soluble solids was not different at harvest; meaning that ELR decreased the partitioning of carbohydrates to fruit. Anthocyanins and flavonols were enhanced by PB-ME, however neither ABA nor ethylene were similarly altered. Instead, the leaf area at nodes above the fruit-zone was lower in PB-ME compared to non-defoliated ones, which increased post-veraison fruit temperature (+2.8 °C). These parameters correlated with anthocyanins at harvest. In conclusion, skin phenylpropanoid concentrations were influenced by canopy density above the fruit-zone. Additionally, ripening-related phytohormones were not involved in the response of phenylpropanoid biosynthesis in vine subjected to ELR.

Kaolin Reduces ABA Biosynthesis Through the Inhibition of Neoxanthin Synthesis in Grapevines Under Water Deficit.

In many viticulture regions, multiple summer stresses are occurring with increased frequency and severity because of warming trends. Kaolin-based particle film technology is a technique that can mitigate the negative effects of intense and/or prolonged drought on grapevine physiology. Although a primary mechanism of action of kaolin is the increase of radiation reflection, some indirect effects are the protection of canopy functionality and faster stress recovery by abscisic acid (ABA) regulation. The physiological mechanism underlying the kaolin regulation of canopy functionality under water deficit is still poorly understood. In a dry-down experiment carried out on grapevines, at the peak of stress and when control vines zeroed whole-canopy net CO2 exchange rates/leaf area (NCER/LA), kaolin-treated vines maintained positive NCER/LA (~2 µmol m-2 s-1) and canopy transpiration (E) (0.57 µmol m-2 s-1). Kaolin-coated leaves had a higher violaxanthin (Vx) + antheraxanthin (Ax) + zeaxanthin (Zx) pool and a significantly lower neoxanthin (Nx) content (VAZ) when water deficit became severe. At the peak of water shortage, leaf ABA suddenly increased by 4-fold in control vines, whereas in kaolin-coated leaves the variation of ABA content was limited. Overall, kaolin prevented the biosynthesis of ABA by avoiding the deviation of the VAZ epoxidation/de-epoxidation cycle into the ABA precursor (i.e., Nx) biosynthetic direction. The preservation of the active VAZ cycle and transpiration led to an improved dissipation of exceeding electrons, explaining the higher resilience of canopy functionality expressed by canopies sprayed by kaolin. These results point out the interaction of kaolin with the regulation of the VAZ cycle and the active mechanism of stomatal conductance regulation.

Exogenous melatonin improves glutathione content, redox state and increases essential oil production in two Salvia species under drought stress.

This research was conducted to understand the influence of foliar applied melatonin (0, 50, 100, 150 and 200 µM) on two Salvia species (Salvia nemorosa L., and Salvia reuterana Boiss) under conditions of water stress. Water stress was applied using a reduced irrigation strategy based on re-watering at 80%, 60% and 40% of the field capacity (FC). Increasing water stress, while significantly enhancing malondialdehyde (MDA), H2O2, electrolyte leakage, oxidized glutathione (GSSG), and total glutathione (GT), reduced glutathione (GSH), catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and glutathione reductase (GR) activities, which led to a marked reduction in fluorescence (Fv/Fm). Foliar application of melatonin alleviated the oxidative stress by increasing GT, CAT, POD, SOD and GR activities and reducing GSSG. In particular, melatonin heightened GSH content as well as the ratio of GSH/GSSG when compared to non-sprayed water stressed plants. Melatonin-treated plants had significantly lower SOD and POD activities than control plants under drought stress, while the CAT activity was enhanced with the foliar treatment. Essential oil yield of both Salvia species increased with the decrease in irrigation from 80% to 60% FC but diminished with the more severe water deficit (40% FC). Essential oil components of Salvia nemorosa were ß- caryophyllene, germacrene- B, spathulenol, and cis- ß- farnesene, while (E) - ß- ocimene, α- gurjnnene, germacrene-D, hexyl acetate and aromadendrene was the major constituents of Salvia reuterana. When plants were subjected to water deficit, melatonin treatment increased the concentration and composition of the essential oil. In particular, melatonin treatments improved the primary oil components in both species when compared to non-melatonin treated plants. In conclusion, reduced irrigation regimes as well as melatonin treatments resulted in a significant improvement of essential oil production and composition in both Salvia species.

Analysis of Non-Structural Carbohydrates and Xylem Anatomy of Leaf Petioles Offers New Insights in the Drought Response of Two Grapevine Cultivars.

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.

Impacts of Pre-bloom Leaf Removal on Wine Grape Production and Quality Parameters: A Systematic Review and Meta-Analysis.

Wine grape (Vitis vinifera L.) is the most widely cultivated fruit crop in the world. However, the climactic characteristics in some growing regions are suboptimal for grape production, including short season length and excess precipitation. Grape growers can utilize an array of methods to mitigate these issues, including "early leaf removal," a management practice involving the removal of leaves from selected basal nodes along shoots around bloom. This meta-analysis reviews the extensive literature on this practice, with specific regards to application at "pre-bloom" (PB). One hundred seventy-five publications on the topic of "early leaf removal" were identified using key terms and subsequently narrowed via eight data curation steps. The comparison between treated (PB) and control plants in these studies revealed two important results. First, PB lowered bunch rot disease (-61%), partially through reducing the compactness of clusters. Second, PB promoted a significant increase in fruit total soluble solids (°Brix, +5.2%), which was related to the increase in the leaf-to-fruit ratio. Furthermore, cultivar and rootstock were found to have a large influence on the success of PB, while the contribution of climate was smaller. In conclusion, PB significantly lowers yield and bunch rot disease and increases °Brix, both of which improve grape and wine quality.

Increase in seed tannin extractability and oxidation using a freeze-thaw treatment in cool-climate grown red (Vitis vinifera L.) cultivars.

Grape seed maturation involves the gradual oxidation of tannins, decreasing excessive bitterness and astringency in wine. In cool climates, this process is limited by the short growing season, affecting wine quality. A "freeze-thaw" treatment on seeds of red vinifera cultivars at veraison and harvest was used to evaluate the effect of oxidation and extractability on seed phenolic fractions. Freezing increased the extraction of total phenolics and o-diphenols quantified from fractionation (fraction 1, vacuolar tannins; fraction 2, hydrogen bonded tannins; fraction 3, covalently bonded tannins), especially at harvest. Despite this, colorimetry, microscopy, oxidation reactivity index (ORI), and correlations between the color index and fractions indicated that freezing disrupted vacuole integrity, enhancing oxidation in the seed coat. In conclusion, vacuolar tannins (which are the main seed phenolics extracted during fermentation) were highly correlated with seed color change, potentially providing information for winemaking in cool climate regions.

Understanding kaolin effects on grapevine leaf and whole-canopy physiology during water stress and re-watering.

Kaolin applications have been investigated in grapevines to understand cooling effects on leaves and clusters and the relative impact on gas exchange, leaf biochemistry, water use efficiency, glyco-metabolism and hormonal patterns. Several Almost all previous contributions have relied upon single-leaf measurements, leaving uncertainty on whole canopy performances, depending on the complexity of a canopy system vs. individual leaves. In our study, kaolin was sprayed at pre-veraison (DOY 204) on potted mature vines (cv. Sangiovese) and washed off a month later (DOY 233), while control vines were left unsprayed. Within control (C) and kaolin (KL) treated vines, well-watered (WW) and water stress (WS) treatments were also imposed over a 10-day period (DOY 208-217) and all vines were re-watered when the WS reached its peak (stem water potential between -1.3 and -1.6 MPa). Single leaf measurements included leaf surface temperature by thermal imaging (Leaf Tmean), assimilation (Leaf A), transpiration (Leaf E), stomatal conductance (Leaf gs) rates, Fv/Fm fluorescence ratio, pre-dawn and stem water potential. Concurrently, whole canopy gas exchange was monitored continuously from DOY 200-259 using a vine enclosure system and daily net CO2 exchange rate (NCER) and canopy transpiration (Ecanopy) were calculated and then normalized vs. leaf area per vine. Results report that for any of the parameters recorded at both levels (single leaf and whole canopy), there was good agreement in terms of relative changes. In absence of water stress, KL was able to improve leaf cooling, while slightly reducing photosynthetic and water loss rates. More interestingly, data taken under water deficit and upon re-watering support the hypothesis that KL can turn into a protective agent for leaf function. In fact, the lack of photo-inhibition and the maintenance of leaf evaporative cooling found in KL-WS at the peak of water-stress (Fv/Fm > 0.7, Leaf Tmean < 38°C and Ecanopy > 0.5mmol m-2 s-1) warranted a prompter recovery of leaf functions upon re-watering that did not occur in C-WS vines.

Metabolic and transcriptional changes associated with the use of Ascophyllum nodosum extracts as tools to improve the quality of wine grapes (Vitis vinifera cv. Sangiovese) and their tolerance to biotic stress.

BACKGROUND: Recent studies report that Ascophyllum nodosum extracts, once applied on the canopy of different crops, deliver positive effects, increasing yield, inducing tolerance to biotic stress, and improving the quality of products. However, the mechanisms of action are still unclear. In this research, vines subjected to multiple foliar applications of an A. nodosum extract (ANE) at label doses were compared with untreated vines (NTV) in accordance with a comparative approach. The investigation coupled a field experiment with a second trial conducted under semi-controlled conditions, to clarify the mechanisms of action involved. RESULTS: The biostimulant did not affect soluble solids or the acidity of grapes; instead, it improved their anthocyanin and phenolic concentrations and the respective profiles. At the time of harvest, anthocyanin, and phenolic concentration were increased by 10.4% and 14.5%, respectively, when compared to the NTV. These effects correlated with a specific modulation of genes involved in the flavonoid metabolic pathways. Moreover, grapes from ANE vines witnessed a significant reduction in the spreading of gray mold when they were either assessed in field conditions or in vitro, compared to the grapes of NTV vines. This was related to a significant upregulation of the defense-related genes of the plant. CONCLUSIONS: Overall, the results showed that A. nodosum extracts can be valuable tools in viticulture considering the emergence of challenging environmental conditions; hence, the regulation of specific metabolic pathways is the mechanism of action that leads to an increased tolerance of biotic stress and of changes in the content of grape metabolites. © 2019 Society of Chemical Industry.

Changes in Within-Shoot Carbon Partitioning in Pinot Noir Grapevines Subjected to Early Basal Leaf Removal.

Early leaf removal significantly alters the source-sink balance within grapevine shoots, leading to a reduction in fruit set. However, no research has previously examined the conditions controlling this process in terms of carbon allocation among major sink organs following defoliation. In this study, the impact of defoliation at bloom on the distribution dynamics of leaf assimilates among clusters and growing shoot apices was investigated on Vitis vinifera, cv. Pinot noir, grown in Michigan, a cool climate viticultural region. Three levels of defoliation: no leaves removed (LR-0); six leaves removed from six basal nodes (LR-6); and ten leaves removed from ten basal nodes (LR-10), were imposed at full bloom. A 13C pulsing was performed 1 week after the treatment application to the defoliated shoots. Single leaf gas exchange (Pn), diurnal changes of the leaf net CO2 assimilation rate, carbon distribution, fruit-set, yield, and fruit composition were measured. Higher Pn was recorded in diurnal measurements of gas exchange in leaf removal (LR) treatments compared to LR-0. The shoot apex of LR-10 experienced the highest 13C allocation (%) after 3 and 7 days following the carbon pulsing. LR-10 had lower percentage of 13C allocated to clusters, which decreased fruit set by 60%, compared to the control, and enhanced the concentration of phenolic compounds in fruit. Alteration of carbon portioning among shoot sink organs indicated that an increasing severity of leaf removal significantly reduced fruit set, and was linearly correlated to shoot apex sink strength, which occurred at the expense of the cluster.

Enhancement of Fruit Technological Maturity and Alteration of the Flavonoid Metabolomic Profile in Merlot ( Vitis vinifera L.) by Early Mechanical Leaf Removal.

Removal of basal leaves near blooms inevitably affects grapevine balance and cluster microclimate conditions, improving fruit quality. Mechanization of this practice allows growers to save time and resources, but to our knowledge, it has not yet been compared with the manual application of this practice in a cool-climate region where seasonal temperatures frequently limit fruit technological maturity and phenolic ripening in red Vitis vinifera cultivars. In our research, berry sugar concentration was highest with prebloom mechanical treatment (PB-ME). Furthermore, metabolomics analysis revealed that PB-ME favored the accumulation of significantly more disubstituted anthocyanins and flavonols and OH-substituted anthocyanins compared with manual application. Given that vine balance was similar between treatments, increased ripening with PB-ME is likely due to enhanced microclimate conditions and higher carbon partitioning through a younger canopy containing basal leaf fragments proximal to fruit. This information provides an important strategy for consistently ripening red Vitis vinifera cultivars in cool climates.

Utilization of a freeze-thaw treatment to enhance phenolic ripening and tannin oxidation of grape seeds in red (Vitis vinifera L.) cultivars.

Phenolic ripening represents a major interest for quality wine producers. Nevertheless, climatic or genotypical limitations can often prevent optimal maturation process. During winemaking seeds can be easily separated and technologically processed to improve their quality. Relying on the key role of oxidation for phenolic ripening, a freeze-thaw treatment was proposed to improve the fruit quality for potential use in challenging growing conditions. The experiment was carried on in two distinctive viticultural areas, Michigan and Italy. Five cultivars (Cabernet Franc, Cabernet Sauvignon, Merlot, Pinot noir and Chambourcin) and six cultivars (Cabernet Sauvignon, Sangiovese, Syrah, Croatina, Barbera and Nebbiolo) were used in Michigan and Italy, respectively. Samples were collected at different phenological stages, to describe the natural ripening process and grape seeds were characterized before and after a freeze-thaw treatment. Colorimetric and spectrophotometric data highlighted similarities among natural and artificial seed ripening promising future applications for the wine industries.