Berries Of Vitis Vinifera Research Articles

Loss ofDiel Circadian Clock Gene Cycling Is a Part ofGrape Berry Ripening.

Diel cycles of gene expression are thought to adapt plants to 24-h changes in environmental conditions. The circadian clock contributes to this process, but less is known about circadian programs in developing reproductive organs. While model plants and controlled conditions have contributed greatly to our knowledge of circadian clock function, there is a need to better understand its role in crop plants under field conditions with fluctuating light and temperature. In this study, we investigated changes in the circadian clock during the development of grape berries of Vitis vinifera L. We found that the transcripts of circadian clock homologs had high-amplitude oscillations prior to, but not during, ripening. As ripening progressed, the amplitude and rhythmicity of the diel oscillations decreased until most transcripts tested had no significant fluctuation over the 24-h cycle. Despite this loss of rhythmicity, the majority of circadian clock genes investigated were expressed at or near their abundance at the nadir of their pre-ripening oscillation although the berries remained transcriptionally active. From this, it can be concluded that cycling of the canonical circadian clock appears unnecessary for berry ripening. Our data suggest that changes in circadian clock dynamics during reproductive organ development may have important functional consequences.

A novel approach to discriminate the volatilome of Vitis vinifera berries by selected ion flow tube mass Spectrometry analysis and chemometrics

This study investigated a quick way to discriminate grape varieties based on their composition in volatile compounds through a SIFT-MS scan coupled with simple chemometrics approaches such as analysis of variance (ANOVA), principal component analysis (PCA) and hierarchical ascendant classification (HAC).The 23 studied grape varieties were distinguishable using O2+, H3O+ and NO+ as reagent ions, and the combination of these three ions. For its ability to ionize most compounds, to efficiently fragment them to generate ions with distinct m/z ratio, and to enhance the differentiation of compounds of similar masses, O2+ reagent ion should be preferentially considered. The use of one single ion rather than three enables to limit the time of analysis and the number of variables to be treated. The technique allowed the distinction of high and low aroma compounds producers as confirmed by headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (HS-SPME/GC-MS) analyses.SIFT-MS is a quick and interesting tool with potential application in various fields of viticulture such as phenotyping of grape varieties or non-targeted studies on the impact of environmental factors or viticultural practices on grape aroma composition.

Extraction and Analysis of Phenolic Compounds from Grape Berries.

Phenolics are ubiquitous compounds that represent the most abundant and diverse class of plant metabolites. From an analytical point of view, phenolics can be divided into soluble phenolics such as phenolic acids, phenylpropanoids, flavonoids and quinones, and nonsoluble compounds such as proanthocyanidins, lignins, and cell wall-bound hydroxycinnamic acids. Extraction of phenolics from the sample material is the first step toward their analysis. Biochemical methods for determination of total phenolics content were widely used in the past but modern chromatographic and mass spectrometric methods for identification and quantification of individual compounds are available in recent years. In this chapter, we describe methods for phenolic compounds extraction used in our laboratories from berries of Vitis vinifera and analytical methods including HPLC coupled to DAD detector and Q-TOF LC/MS for their analysis.

Implication of Row Orientation Changes on Fruit Parameters of Vitis vinifera L. cv. Riesling in Steep Slope Vineyards.

Row orientation, among others, is a crucial factor in determining grapevine performance and health status, thus affecting berry components that form the basis of the later wine profile. However, the literature about the impact of changes in row orientation at steep slope sites on grapevine fruit composition as well as the differentiation between canopy sides hardly exists. Thus, the aim of this work was to gain knowledge about the impact of row orientation in steep slope vineyards on selected primary and secondary metabolites in berries of Vitis vinifera L. cv. Riesling. Samples were taken from both canopy sides of different row orientations of terraced and downslope vineyards in steep slopes. Free amino acids in the juice and flavonols in the berry skin had a positive correlation to sunlight exposure. Furthermore, grapevines showed adaptations to constantly higher light conditions, e.g., physiologically in reduction in chlorophyll content or protective mechanisms resulting in a lower susceptibility to sunburn damage. Thus, grapevine fruit parameters are affected by row orientation change in steep slopes.

Melatonin confers enhanced polyamine metabolism and cell tolerance in Vitis vinifera against oxidative damage: Quantitative proteomic evidence

Melatonin (MLT) exhibited pleiotropic effects in multiple plant physiological processes; however, its role in the polyamine metabolism and quality performance of postharvest Vitis vinifera berry remain unknown. In the present study, MLT was applied to postharvest Vitis vinifera cv. Kyoho berry, which delayed berry abscission, berry rotten, and promoted cell tolerance to oxidative stress. Notably, the malondialdehyde content of berries was reduced to 40.8 % on the application of 200 μmol L−1 MLT. Furthermore, tandem mass tags (TMT)-based quantitative proteomic analysis identified 5,156 proteins from berry skin, among which 158 proteins showed quantitatively differences in response to MLT application. These differentially expressed proteins were mainly upregulated in the arginine and proline metabolism pathways, lysine degradation, and ascorbate and aldarate metabolism pathways. The abundance of proteins involved in polyamine metabolisms, including N-carbamoylputrescine amidase, spermidine synthase, and aldehyde dehydrogenases were also upregulated. Moreover, MLT application enhanced contents of arginine, proline, 4-aminobutanoic acid, and polyamines, which played an essential role in oxidative stress response. Further, the transcript level of key enzymes involved in polyamine metabolism, including VvADC, VvODC, VvNAC, VvSPDS and VvCuAO were up-regulated on MLT application. Overall, the role of MLT in regulating the polyamine metabolism and subsequently in cell tolerance against oxidative damage was scientifically established for the first time for Vitis vinifera. The empirical findings of this study may propose new insights on MLT application in postharvest berry physiology and provide theoretical regulation for the preservation of berries, perhaps other fruits.

Profiling Analysis of Volatile and Non-volatile Compounds in Vitis Vinifera Berries (cv. Chardonnay) and Spontaneous Bud Mutation

A novel clonal variety of Vitis vinifera was identified from “Chardonnay” using inter-simple sequence repeat (ISSR) markers and called “bud mutation. ” The metabolomic profiles in Chardonnay and bud mutation berries indicated essential differences in the expression of key genes in the pathways of 2-C-methyl-D-erythritol-4-phosphate (MEP) and lipoxygenase-hydroperoxide lyase (LOX-HPL). Bud mutation fruits also matured 10 days earlier than Chardonnay and have higher carotenoid, sugar, and acidic compound contents. Furthermore, the gene expression was examined in the biosynthetic pathways of two ripening-associated hormones, abscisic acid (ABA) and jasmonic acid (JA), which significantly increased in bud mutation compared with the Chardonnay fruit. The synthesis and metabolism of amino acids, terpenes, fatty acids, volatile components, and specialized metabolites significantly increased in bud mutation. Therefore, in comparison with Chardonnay, bud mutation is considered a highly aroma-producing grape variety for an improvement in the beverage industry.

3D visualisation of voids in grapevine flowers and berries using X‐ray micro computed tomography

Background and Aim X-ray micro computed tomography (micro-CT) is a non-destructive 3D imaging technique that has been applied to plant morphology and anatomical studies to gain a better understanding of physiological phenomena in vivo. It is particularly useful for imaging voids in undisturbed fragile tissues and therefore may be applied to the delicate flowers and soft berries of Vitis vinifera. The characterisation of gas spaces and channels can offer insights into the process of tissue aeration and this may have implications on cell function and vitality. We assessed the use of micro-CT to visualise voids within these reproductive organs. Methods and Results The internal structures of flowers and berries were captured through rapid micro-CT scanning and subsequently were recreated in 3D using image processing. The relative positions of the developing flower parts encased within the flower cap were visualised. Low density/porous tissue was identified within the pedicel and receptacle, connecting the lenticels with the interior of the berry. Voids were present in the proximal mesocarp of mature berries forming a ‘detachment zone’ in both seeded and seedless cultivars. Voids permeated the mesocarp of mature seedless grape cultivars, but not seeded grapes. Conclusion Micro-CT offers new insights regarding the distribution of voids on the morphology and compositional heterogeneity of organs that are difficult to dissect and/or view with light microscopy. Significance of the Study A better understanding of the physiology and functionality of grapevine reproductive tissues may be achieved by 3D visualisation of internal structure in vivo.

Assessing macro- (P, K, Ca, Mg) and micronutrient (Mn, Fe, Cu, Zn, B) concentration in vine leaves and grape berries of vitis vinifera by using near-infrared spectroscopy and chemometrics

Macronutrients (phosphorus, potassium, calcium, and magnesium) and micronutrients (manganese, iron, copper, zinc, and boron) play an essential role not only in the general physiology of vines but also in the quality of wine produced. The quantity of each nutrient in the vine is generally determined by analyzing the leaf blades or petioles, but this approach imposes a typical delay of two weeks between sampling and receiving the results, which precludes real-time detection of nutritional deficiencies (e.g., boron deficiency at flowering). Therefore, a method to rapidly analyze vine organs is highly desirable. One candidate for such a method is near-infrared (NIR) reflectance spectroscopy coupled with chemometric methods, based on which winegrowers have already developed prediction models. This approach is widely used today in agriculture. The aim of the present study is to determine whether NIR spectroscopy can be used to obtain accurate information about the nutritional status of vines. In this study, we focus on the mass of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), and boron (B) contained in different vine organs (leaf blades, petioles and berries) over the course of a year. The concentration of these elements was determined based on NIR absorbance spectra from 677 samples of various dried vine organs. Partial least square models for classification and prediction were then developed based on raw and pretreated spectra for each organ, following which the models were tested on an external prediction set. The results show that, for Ca and Mg, all organ models can be used routinely for classification or prediction. For prediction, the Ca (Mg), model produces r2 = 0.88, 0.70, and 0.72 (0.60, 0.72, and 0.80) for leaf blades, petioles, and berries, respectively. Only for leaf blades (berries) is the Ca (Mg) model sufficiently accurate to be used for prediction. For berries, the P, K, and Zn models produce r2 in prediction of 0.77, 0.79, and 0.82, respectively. For petioles, the K model proves reliable for prediction, with r2 = 0.76. The Fe, Cu, and B models produce r2 = 0.72, 0.71, and 0.52, which are suitable for classification but not for prediction. Finally, for leaf blades, the Fe and Cu models produce r2 0.58 and 0.61, respectively, in prediction and thus can be used routinely for classification.

Cell wall and metabolite composition of berries of Vitis vinifera (L.) cv. Thompson Seedless with different firmness

Firm berries are highly appreciated by table grape consumers. Cell wall composition is one of the main factors influencing the firmness of table grape berries. Nevertheless, the biological factors driving changes in berry firmness remain unclear. In the present work, we evaluated the firmness of berries of Vitis vinifera cv. Thompson Seedless. We selected two orchards displaying contrasting berry firmness and evaluated polar metabolites and cell wall composition. Our results suggest that berries from the soft orchard exhibited a higher accumulation of sugars at veraison whereas berries from the hard orchard accumulated the same sugars at harvest plus a higher amount of glucose monosaccharide at the cell wall. Thus, this study opens new insights about a connection between metabolic and cell wall changes with fruit firmness in a table grape variety, suggesting that it is possible to use metabolomic tools to identify metabolic biomarkers associated with table grape berry firmness.

Quantitating Organoleptic Volatile Phenols in Smoke-Exposed Vitis vinifera Berries.

Accurate methods for quantitating volatile phenols (i.e., guaiacol, syringol, 4-ethylphenol, etc.) in smoke-exposed Vitis vinifera berries prior to fermentation are needed to predict the likelihood of perceptible smoke taint following vinification. Reported here is a complete, cross-validated analytical workflow to accurately quantitate free and glycosidically bound volatile phenols in smoke-exposed berries using liquid-liquid extraction, acid-mediated hydrolysis, and gas chromatography-tandem mass spectrometry. The reported workflow addresses critical gaps in existing methods for volatile phenols that impact quantitative accuracy, most notably the effect of injection port temperature and the variability in acid-mediated hydrolytic procedures currently used. Addressing these deficiencies will help the wine industry make accurate, informed decisions when producing wines from smoke-exposed berries.

Manipulating rainfall to study symptom expression of <i>Botrytis cinerea</i> infection in wine grapes

Manipulating rainfall to study symptom expression of <i>Botrytis cinerea</i> infection in wine grapes

Volatiles from Botrytis cinerea‐infected and healthy berries of Vitis vinifera influence the oviposition behaviour of Epiphyas postvittana

AbstractInteractions among plants, plant‐feeding insects, and plant–pathogenic fungi are partially mediated by volatile compounds. Herbivorous insects use sensory cues to choose host plants for feeding and/or oviposition that are likely to support survival and development of progeny. It is known that some fungus‐induced alterations in plants can modify plant volatiles, which are recognized by the olfactory receptors of the insect, either as an attractant or as a deterrent. We tested for the presence of behaviour‐modifying volatiles emanating from the berries of Vitis vinifera L. (Vitaceae) infected with Botrytis cinerea Pers. (Helotiales). We tested the olfactory behaviour of adults of Epiphyas postvittana Walker (Lepidoptera: Tortricidae) to these volatiles using two‐choice and wind‐tunnel experiments. We hypothesized that olfactory cues influence E. postvittana's oviposition behaviour. We found that volatiles emanating from B. cinerea‐infected berries did not significantly attract the gravid females of E. postvittana; consequently, they laid significantly fewer eggs on infected berries. Furthermore, significantly fewer females of E. postvittana were found attracted to infected berries in the wind tunnel assay. Ethanol and 3‐methyl‐1‐butanol were abundant in B. cinerea‐infected berries. Oviposition assays made with laboratory standards of ethanol and 3‐methyl‐1‐butanol confirmed their role in regulating the olfactory behaviour of E. postvittana site selection.

The effects of light and ethylene and their interaction on the regulation of proanthocyanidin and anthocyanin synthesis in the skins of Vitis vinifera berries

Several environmental stimulants and hormones can affect phenolic accumulation in grape berries, but the effects of ethylene and light and their interaction have not been completely deciphered. We measured the accumulation of anthocyanins (ATs) and proanthocyanidins (PAs) in berries of a red-wine grape, Vitis vinifera cv. Cabernet Sauvignon, in four treatments: light exposure with and without ethephon, a commercial growth regulator that quickly converts to ethylene, and box-shading with and without ethephon. Ethephon did not significantly increase the weight and diameter of the berries but altered maturity parameters at 49 days after treatment. Ethephon and light increased the fresh and dry weights of the berry skins, induced the accumulation of total phenolics, total flavonoids, and PAs and up-regulated the transcription of VvLAR1, VvLAR2, VvANR, and VvMYBPA1. Light promoted the accumulation of ATs and the expression of AT biosynthetic genes more effectively than did ethephon. Ethephon and light had interactive effects on grape ripening and the contents of PAs and ATs by the way that light interfered with the effect of exogenous ethephon on endogenous ethylene biosynthesis and the downstream ethylene regulatory pathway. These findings provide new information about the physiological and molecular function of ethylene during berry development and the impact of the interaction between ethylene and light in Vitis vinifera.

Transcriptome and metabolome reprogramming in Vitis vinifera cv. Trincadeira berries upon infection with Botrytis cinerea.

Vitis vinifera berries are sensitive towards infection by the necrotrophic pathogen Botrytis cinerea, leading to important economic losses worldwide. The combined analysis of the transcriptome and metabolome associated with fungal infection has not been performed previously in grapes or in another fleshy fruit. In an attempt to identify the molecular and metabolic mechanisms associated with the infection, peppercorn-sized fruits were infected in the field. Green and veraison berries were collected following infection for microarray analysis complemented with metabolic profiling of primary and other soluble metabolites and of volatile emissions. The results provided evidence of a reprogramming of carbohydrate and lipid metabolisms towards increased synthesis of secondary metabolites involved in plant defence, such as trans-resveratrol and gallic acid. This response was already activated in infected green berries with the putative involvement of jasmonic acid, ethylene, polyamines, and auxins, whereas salicylic acid did not seem to be involved. Genes encoding WRKY transcription factors, pathogenesis-related proteins, glutathione S-transferase, stilbene synthase, and phenylalanine ammonia-lyase were upregulated in infected berries. However, salicylic acid signalling was activated in healthy ripening berries along with the expression of proteins of the NBS-LRR superfamily and protein kinases, suggesting that the pathogen is able to shut down defences existing in healthy ripening berries. Furthermore, this study provided metabolic biomarkers of infection such as azelaic acid, a substance known to prime plant defence responses, arabitol, ribitol, 4-amino butanoic acid, 1-O-methyl- glucopyranoside, and several fatty acids that alone or in combination can be used to monitor Botrytis infection early in the vineyard.

Metabolic changes of Vitis vinifera berries and leaves exposed to Bordeaux mixture

Since the development of Bordeaux mixture in the late 1800's, copper-based fungicides have been widely used against grapevine (Vitis vinifera L.) diseases, mainly in organic but also in conventional viticulture; however their intensive use has raised phytotoxicity concerns. In this study, the composition of grape berries and leaves upon Bordeaux mixture treatment was investigated during the fructification season by a metabolomic approach. Four applications of Bordeaux mixture till 3 weeks before harvest were performed following the regular management practices of organic viticulture. Results showed that the copper-based treatment affected the content in sugars, organic acids, lipids and flavan-3-ols of grapes and leaves at specific developmental stages. Nonetheless, the levels of sucrose, glucose and fructose, and of tartaric and malic acids were not significantly affected in mature grapes. In contrast, a sharp decrease in free natural amino acids was observed, together with a reduction in protein content and in mineral nitrogen forms. The treatment with Bordeaux mixture increased by 7-fold the copper levels in tissue extracts from surface-washed mature berries.

Solute accumulation differs in the vacuoles and apoplast of ripening grape berries

Phloem unloading is thought to switch from a symplastic route to an apoplastic route at the beginning of ripening in grape berries and some other fleshy fruits. However, it is unclear whether different solutes accumulate in both the mesocarp vacuoles and the apoplast. We modified a method developed for tomato fruit to extract apoplastic sap from grape berries and measured the changes in apoplastic and vacuolar pH, soluble sugars, organic acids, and potassium in ripening berries of Vitis vinifera 'Merlot' and V. labruscana 'Concord'. Solute accumulation varied by genotype, compartment, and chemical species. The apoplast pH was substantially higher than the vacuolar pH, especially in Merlot (approximately two units). However, the vacuole-apoplast proton gradient declined during ripening and in Merlot, but not in Concord, collapsed entirely at maturity. Hexoses accumulated in both the vacuoles and apoplast but at different rates. Organic acids, especially malate, declined much more in the vacuoles than in the apoplast. Potassium accumulated in the vacuoles and apoplast of Merlot. In Concord, by contrast, potassium increased in the vacuoles but decreased in the apoplast. These results suggest that solutes in the fruit apoplast are tightly regulated and under developmental control.

Increase of Fumonisin B2 and Ochratoxin A Production by Black Aspergillus Species and Oxidative Stress in Grape Berries Damaged by Powdery Mildew

Powdery mildew (PM), caused by the fungus Erysiphe necator, is one of the most widespread fungal disease of grape and may cause extensive openings on the berry surface during the infection. We evaluated the effect of damage caused by PM in grape berries on the growth of and mycotoxin production by Aspergillus and on the oxidative stress in infected berries. Berries of Vitis vinifera L. cv. Negroamaro with sound skin (SS) and those naturally infected by PM were surface sterilized and inoculated with either fumonisin B2(FB2)-producing strains of Aspergillus niger or ochratoxin A (OTA)-producing strains of Aspergillus carbonarius and incubated at 20 and 30°C. The PM berries were significantly more susceptible to both Aspergillus colonization (5 to 15 times more susceptible) and OTA and FB2 contamination (2 to 9 times more susceptible) than were SS berries. The highest toxin concentration was detected in inoculated PM berries both for OTA (9 ng/g) at 20°C and for FB2 (687 ng/g) at 30°C. In inoculated SS and PM berries, although malondialdehyde and hydrogen peroxide concentrations did not increase, the two black Aspergillus species caused a significant decrease in ascorbate content, thus inducing a pro-oxidant effect. These results indicate that grape berries affected by PM are more susceptible to black Aspergillus growth and to production and/or accumulation of FB2 and OTA. Thus, preventive control of E. necator on grape berries could reduce the mycotoxicological risk from black Aspergillus infection.

The vacuolar channel VvALMT9 mediates malate and tartrate accumulation in berries of Vitis vinifera

Vitis vinifera L. represents an economically important fruit species. Grape and wine flavour is made from a complex set of compounds. The acidity of berries is a major parameter in determining grape berry quality for wine making and fruit consumption. Despite the importance of malic and tartaric acid (TA) storage and transport for grape berry acidity, no vacuolar transporter for malate or tartrate has been identified so far. Some members of the aluminium-activated malate transporter (ALMT) anion channel family from Arabidopsis thaliana have been shown to be involved in mediating malate fluxes across the tonoplast. Therefore, we hypothesised that a homologue of these channels could have a similar role in V. vinifera grape berries. We identified homologues of the Arabidopsis vacuolar anion channel AtALMT9 through a TBLASTX search on the V. vinifera genome database. We cloned the closest homologue of AtALMT9 from grape berry cDNA and designated it VvALMT9. The expression profile revealed that VvALMT9 is constitutively expressed in berry mesocarp tissue and that its transcription level increases during fruit maturation. Moreover, we found that VvALMT9 is targeted to the vacuolar membrane. Using patch-clamp analysis, we could show that, besides malate, VvALMT9 mediates tartrate currents which are higher than in its Arabidopsis homologue. In summary, in the present study we provide evidence that VvALMT9 is a vacuolar malate channel expressed in grape berries. Interestingly, in V. vinifera, a tartrate-producing plant, the permeability of the channel is apparently adjusted to TA.

Effects of Water-Deficit Irrigation on Hormonal Content and Nitrogen Compounds in Developing Berries of Vitis vinifera L. cv. Tempranillo

Water-deficit irrigation to grapevines reduces plant growth, yield, and berry growth, altering the ripening process, all of which may influence fruit composition and wine quality. Therefore, the goals of this study were (1) to investigate the influence of the main endogenous berry hormones, abscisic acid (ABA), indole-3-acetic acid (IAA), salicylic acid (SA), and jasmonic acid (JA), on berry growth and ripening under water-deficit conditions and (2) to analyze changes in fruit composition, specifically N compounds, under water deprivation. The study was carried out using container-grown Tempranillo grapevines grown under controlled conditions in a greenhouse. Two irrigation treatments were imposed: control (well-watered) and sustained deficit irrigation (SDI). Water deficit decreased leaf area and the source-to-sink ratio, reduced yield and berry size, and decreased concentrations of the main phenolic compounds. SDI also modified berry hormonal status. At the pea-size stage, SDI berries had lower IAA and higher JA and SA than nonstressed berries. At veraison (onset of ripening), accumulation of ABA was less accentuated in SDI than in control berries. At harvest, the content of amino acids and free ammonium was low in both treatments but SDI-treated berries showed a significant accumulation of amines. Results suggest that water restrictions to grapevines might be playing a physiological role in reducing berry growth through affecting hormone dynamics, phenolic synthesis, and the berry amino acid content and composition, which could compromise fruit quality. Possible roles of endogenous IAA controlling berry size and endogenous ABA and SA controlling levels of anthocyanins and flavonols at harvest are discussed.

Lachancea lanzarotensis sp. nov., an ascomycetous yeast isolated from grapes and wine fermentation in Lanzarote, Canary Islands

During the characterization of the microbiota biodiversity associated with grapes and wineries in different bioclimatic conditions of the Canary Islands (Spain), a novel yeast species was isolated from Lanzarote, the driest wine-producing region of the archipelago. Seven strains isolated from grapes, microvinifications and wineries are described. Sequence analysis of the D1/D2 domain of the LSU rDNA gene and 5.8S-ITS regions revealed that the isolates were phylogenetically a member of the genus Lachancea and are closely related to Lachancea meyersii NRRL Y-27269(T) and Lachancea nothofagi NRRL Y-48670(T). On the basis of morphological, biochemical and physiological characterization and phylogenetic analysis, a novel ascosporogenous yeast species, Lachancea lanzarotensis sp. nov., is proposed. The type strain is L2C-15(T) ( = CBS 12615(T) = CECT 13066(T)) which was isolated from grape berries of Vitis vinifera L. cv. Listán Negro red grape variety in Tinajo, Lanzarote. The MycoBank no. is MB 801390.