Pesticide Residues and Berry Microbiome after Ozonated Water Washing in Table Grape Storage.

Nowadays, different systems for reducing pesticides in table grapes are being tested at different production stages either in the field or in postharvest. The present study tested ozonated water treatments at the beginning of the cold storage of the Princess® seedless table grape variety to reduce the residue contents of some pesticides and to evaluate their effect on gray mold and the berry microbiome. An ozone generator capable of producing an ozone concentration ranging from 18 to 65 Nm3 was utilized for obtaining three ozone concentration levels in water: 3, 5 and 10 mg/L. Ozonated water was placed in a 70 L plastic box where 500 g grape samples closed in perforated plastic clamshell containers were immersed utilizing two washing times (5 and 10 min). Overall, six ozonated water treatments were tested. After the ozonated water treatments, all samples were stored for 30 days at 2 °C and 95% relative humidity to simulate commercial practices. The pesticide residue contents were determined before the ozonated water treatments (T0) and 30 days after the cold storage (T1). The treatments with ozonated water washing reduced the pesticide residues up to 100%, while the SO2 control treatment reduced the pesticide residues ranging from 20.7 to 60.7%. Using 3 mg/L ozonated water to wash grapes for 5 min represented the optimal degradation conditions for all of the analyzed pesticides, except for fludioxonil, which degraded better with a washing time of 10 min. The ozone treatments did not significantly reduce the gray mold and the fungal and bacterial microbiome, while a relevant reduction was observed in the yeast population.

Effect of over crop and reduced yield by cluster thinning on phenolic and volatile compounds of grapes and wines of 'Sangiovese' trained to Tendone.

BACKGROUND: 'Sangiovese' (Vitis vinifera L.) is a well-known Italian wine grape utilized for producing the prestigious Chianti Classico brand. Here, 'Sangiovese' is generally trained to a vertical training system, with an imposed crop level of around 3-5 kg per vine to preserve grape quality. Tendone comprises a horizontal trellis system, largely diffused in Mediterranean Countries for table grape production, and is adopted to achieve a high yield and to preserve grapes, by means of its canopy architecture, from excessive solar radiation, which in turn affects the organoleptic aspects of grapes and wines. The present study aimed to evaluate, in terms of phenolic and volatile compounds of grape and wines, 'Sangiovese' trained to Tendone, as affected by two crop levels, namely very high and reduced yields. RESULTS: Reduced yield (18.39 kg per vine) by cluster thinning still appeared to provide a considerable yield and quality of grapes and wines. Thinning increased pH, tartaric acid content, flavonoids, anthocyanins, total polyphenols and proanthocyanins, and total free and bound aroma compounds, without affecting soluble solids and titratable acidity of grapes. As regards wines, reduced yield increased pH, tartaric and malic acids and dry reduced extract, flavonoids, anthocyanins, total polyphenols and proanthocyanins, and, finally, the color intensity of wines. CONCLUSION: The present study highlights that Tendone training systems, combined with 40% of cluster thinning, allows a high yield to be reached, allowing, at the same time, a very interesting quality of grapes, making it possible to produce qualitative wines. © 2022 Society of Chemical Industry.

Optimized ultrasonic-assisted extraction of phenolic antioxidants from grape (Vitis vinifera L.) skin using response surface methodology.

Currently, the analysis of bioactive compounds is easily carried out by modern chromatographic and spectrometric techniques, but the success of the procedure still depends on the efficiency of the extraction methods. This work aimed at finding the optimal conditions for ultrasonic-assisted extraction (UAE) of phenolic antioxidants from grape skin using single-factor experimentation combined with response-surface methodology. Our extraction parameters were ethanol concentration, time and temperature. Total phenolic content (TPC and TPC-280), 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and oxygen radical absorbance capacity (ORAC) were dependent variables. A second-order polynomial model satisfactorily described the experimental data. Optimized extraction conditions for UAE from grape skin were: 59.5% ethanol, 113.6 min and 66.8 °C for TPC extraction; 61.1% ethanol, 99.4 min and 66.1 °C for TPC-280 extraction; 57.1% ethanol, 99.5 min and 66.8 °C for DPPH radical-scavenging activity; 60.2% ethanol, 82.4 min and 66.8 °C for ORAC value. Our work provides detailed and useful information to optimize the extraction of grape skin phenols, which are widely used as dietary supplements and nutraceuticals.

Effect of two exogenous plant growth regulators on the color and quality parameters of seedless table grape berries.

Some red-pink table grape varieties, cultivated in warm climates, can fail in achieving the right level of anthocyanins responsible for the intense and uniform red color of berries. Nowadays, this is becoming an important technological issue in the Mediterranean area, which may result in decreasing market acceptance and potential economic value of table grape. Usually, plant growth regulators or phytohormones, such as S-ABA, can overcome this problem because they drive the accumulation of anthocyanins over the ripening season. Harpin proteins (HrP), which enhance the plant disease resistance, may be supposed to stimulate the anthocyanins biosynthesis in grape skin if applied close to veraison. Therefore, this research aimed at comparing the effect of HrP and S-ABA over the anthocyanin and color improvement of Crimson Seedless table grape grown in Southern Italy. For the first time, the exogenous treatment with HrP showed as effective as the less sustainable S-ABA one in favoring the anthocyanin accumulation, leading to peonidin-3-O-glucoside, cyanidin-3-O-glucoside, and malvidin-3-O-glucoside values up to 4 folds higher than control grapes and giving rise to a greater concentration of the more stable acylated anthocyanins. Overall, the color of berries was improved but keeping high the other quality characteristics.

Ampelometric Leaf Trait and SSR Loci Selection for a Multivariate Statistical Approach in Vitis vinifera L. Biodiversity Management.

High estimated heritability values were recently revealed for mature leaf traits in grape (Vitis vinifera L.), thus redeeming ampelography in the era of molecular markers. The "Organisation Internationale de la Vigne et du Vin (OIV)" set a list of hundreds of descriptors for grapevine in order to standardize ampelographic and ampelometric scores. Therefore, the selection and reduction of the number of OIV codes can represent a major goal for leaner biodiversity assessment studies. The identification of ampelometric traits associated with grape diversity allows to construct Classification Trees with chi squared automatic interaction detection (CHAID) algorithm, a stepwise model-fitting method that produces a tree diagram in which at each step the sample pool is splitted based on the independent variables statistically different for the dependent variable. A collection of 100 table and wine grapevines (Vitis vinifera L.) was characterized and evaluated by means of six microsatellites and twenty-two ampelometric traits on mature leaves. Nine ampelometric traits were selected by principal component analysis and employed to build the classification trees based on CHAID algorithm. The strategy can represent an effective tool for grape biodiversity management, right allocations, and identification of new grape genotypes, implemented by a further microsatellite investigation only when unsolved cases occur, allowing faster and cheaper results.

Maceration with stems contact fermentation: effect on proanthocyanidins compounds and color in Primitivo red wines.

Three Primitivo (Vitis vinifera, cv.) red wines were microvinified by means of different winemaking technologies: no stem-contact fermentation destemming 100% of grapes (D100); stem-contact fermentation destemming 75%, 50% of grapes (D75-D50) respectively. The objectives of this work were to improve proanthocyanidins content in wine, to monitor the relationships between anthocyanins/tannins and to detect the effects on the polymerization state of polyphenols after 12 months storage of wines. D100 showed higher anthocyanins content but lower color intensity and phenolic compounds content with respect to the theses D75 and D50, the last two showing lower anthocyanins content due their partial adsorption by grape stems during the fermentation. D75 gave the best results in terms of anthocyanins/color intensity balance and showed a better wine tannin component with respect to D50. Moreover after 12 months storage D50 reached a more advanced and stable polymerization state of colored pigments than the other wines.

Morphological variability in leaves and molecular characterization of novel table grape candidate cultivars (Vitis vinifera L.).

The present work report the characterization of twenty-one table grapes candidate cultivars plus five registered ones included as reference, by means of 47 ampelographic traits, 23 ampelometric measurements and six microsatellite loci. The final goal of the research was to analyse the possibility of reducing the number of morphological and molecular tools required for a precise and effective description of a grape genotype or cultivar. This would be of great help for future biodiversity description on a larger sample of more than 300 table grapes accessions today grown at the 'Consiglio per la Ricerca e la sperimentazione in Agricoltura (C.R.A.)-Unità di ricerca per l'uva da tavola e la vitivinicoltura in ambiente mediterraneo (Bari-Italy)'. OIV ampelographic traits showed a clear distinction among all twenty-six genotypes analysed, suggesting the relevant morphological variability investigated. Principal component analysis based on ampelometric traits revealed main veins ON(3), ON(4) and O(3)N(4); ratios between main veins; angles between main veins and of petiolar sinus, to be the most effective records in differentiating cultivars, for a total variation of 69.9 % described by the first three components. Molecular analysis based on six microsatellite loci was performed on all genotypes, providing a detailed molecular profile and a dendrogram of genetic similarity, in which all genotypes were clearly distinguishable. Finally, with the goal of using the minimum possible number of markers to differentiate genotypes, microsatellites VVMD5 and VVMD27 were selected to be sufficient to distinguish among all the candidate cultivars included in the analysis, representing a possible 'step by step' approach when a molecular characterization has to be undertaken on a large number of genotypes, by first testing few markers and increasing their number only if necessary.

Validation assay of p3_VvAGL11 marker in a wide range of genetic background for early selection of stenospermocarpy in Vitis vinifera L.

DNA markers technology, derived from research in molecular biology and genomics, offers great promise for plant breeding, allowing the "molecular breeding" via marker-assisted selection. Grapevine genomic resources allowed, in recent years, the characterization at molecular level of genes involved in interesting phenotypes such as stenospermocarpic seedlessness, a trait really appreciated by consumers. Recent studies in table grapes revealed that the VvAGL11 gene, member of the D-lineage MADS-box family, controls the ovule identity, and thus potentially playing an important role in stenospermocarpy. Intragenic markers of VvAGL11 have been found and tested for breeding purposes. In the present paper, we describe an in deep assay on a total of 475 genotypes derived by our own grape germplasm and seeded × seedless crosses F1 offspring, to evaluate and verify the "diagnostic" power of VvAGL11 in marker-assisted selection. We found only 8/475 that were seeded and carried the seedless-associated allele in the STS p3_VvAGL11. However, and most importantly, there were no seedless varieties without such allele. We validated the marker as a 100 % effective tool for early negative selection of stenospermocarpy in Vitis vinifera L. crosses.

sunTILL: a TILLING resource for gene function analysis in sunflower.

BACKGROUND: Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource. RESULTS: To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M2 progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M2 lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb. CONCLUSION: A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower.

An evaluation of a new approach to the regeneration of Helichrysum italicum (Roth) G. Don, and the molecular characterization of the variation among sets of differently derived regenerants.

A protocol for the induction of regeneration from leaves of Helichrysum italicum was established. Calli were found to form on the basal medium only when it was supplemented with thidiazuron (TDZ) alone or in combination with naphthalene acetic acid (NAA), with a percentage ranking of at least 80%. The hormone-free medium showed the highest percentage of shoot regeneration (62%) even though no callus formed. AFLP markers were employed to verify tissue culture-induced variation in the regenerated plantlets obtained by direct shoot regeneration or the indirect shoot regeneration process (callus formation). Seven out of the eleven AFLP primer pairs yielded polymorphic patterns. The average number of fragments per primer pair was 64.1. Singletons were represented by 12 (2.7%) fragments. Student's T-test was performed both on the average number of shared fragments and on the nucleotide diversity, and no significant statistical difference was observed between the two regeneration treatments.