The positive response in grape secondary metabolites under controlled stresses: a review

Grapevine is cultivated worldwide with great economic importance. In recent years, our knowledge of the physiological and molecular basis of berry quality regulation has substantially increased. Abiotic and biotic stresses, such as deficit irrigation, low temperature, light/UV and microbes, to a certain extent, could improve grape berry quality by enhancing flavor metabolites, colorization or aroma compounds. This review summarizes recent data related to the stress of grape berry development, with special emphasis on secondary metabolism and its response to stresses. A full understanding of how grape berry metabolism responds to different stresses is important to improve the biochemical qualities of grapes and resultant products, such as wine in practice

Metabolic profiling reveals coordinated switches in primary carbohydrate metabolism in grape berry (Vitis vinifera L.), a non-climacteric fleshy fruit

Changes in carbohydrate metabolism during grape berry development play a central role in shaping the final composition of the fruit. The present work aimed to identify metabolic switches during grape development and to provide insights into the timing of developmental regulation of carbohydrate metabolism. Metabolites from central carbon metabolism were measured using high-pressure anion-exchange chromatography coupled to tandem mass spectrometry and enzymatic assays during the development of grape berries from either field-grown vines or fruiting cuttings grown in the greenhouse. Principal component analysis readily discriminated the various stages of berry development, with similar trajectories for field-grown and greenhouse samples. This showed that each stage of fruit development had a characteristic metabolic profile and provided compelling evidence that the fruit-bearing cuttings are a useful model system to investigate regulation of central carbon metabolism in grape berry. The metabolites measured showed tight coordination within their respective pathways, clustering into sugars and sugar-phosphate metabolism, glycolysis, and the tricarboxylic acid cycle. In addition, there was a pronounced shift in metabolism around veraison, characterized by rapidly increasing sugar levels and decreasing organic acids. In contrast, glycolytic intermediates and sugar phosphates declined before veraison but remained fairly stable post-veraison. In summary, these detailed and comprehensive metabolite analyses revealed the timing of important switches in primary carbohydrate metabolism, which could be related to transcriptional and developmental changes within the berry to achieve an integrated understanding of grape berry development. The results are discussed in a meta-analysis comparing metabolic changes in climacteric versus non-climacteric fleshy fruits

Molecular cloning of dihydroflavonol 4-reductase gene from grape berry and preparation of an anti-DFR polyclonal antibody

Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219) is a key enzyme of the flavonoid pathway, which synthesizes numerous secondary metabolites to determine the quality of grape berry and wine. The full-length dfr cDNA with 1014 bp was cloned from grape berry, and then introduced into an expressed plasmid pET-30a (+) vector at the EcoR I and Xho I restriction sites. With induction of the isopropyl-β-D-thiogalactoside (IPTG), the pET-dfr was highly expressed in Escherichia coli BL21 (DE3) pLysS cells. A fusion protein with the His-Tag was purified through Ni-NTA His Bind Resin and then used as the antigen to immunize a New Zealand rabbit. The resulting antiserum was further purified precipitated by 50 % saturated ammonium sulfate and DEAE-Sepharose FF chromatography to obtain the immunoglobulin G (IgG) fraction. The resulting polyclonal antibody was found capable of immuno-recognizing the DFR of the crude protein extracts from grape berry. This work undoubtedly provides the possibility for further studies on biological regulation of DFR activity in grape berry.

A Deep Proteomics Perspective Into Grape Berry Quality Traits During Ripening

Discovery‐based proteomics studies have an important role in the understanding of the biochemical processes that occur during grape berry ripening. The ripening process is relevant in determining grape berry quality. For a proteome analysis of grape berry ripening, Kambiranda et al. (2018) applied a label‐free mass spectrometry–based quantitative approach. The authors reported the identification of proteins associated with the production flavor, aroma and ethylene production. Despite the valuable contribution of discovery‐based proteomics studies, the picture is still incomplete. Future efforts in gaining proteome coverage would benefit the identification of proteins associated with grape berry quality traits

Corrigendum

Ethylene signalling receptors and transcription factors over the grape berry development: gene expression profilingVitis 49 (3), 129-136 (2010

Effects of Abiotic Factors on Phenolic Compounds in the Grape Berry – A Review

Grape berry phenolic compounds are widely described in literature. Phenolics can be divided into two main groups: flavonoids and non-flavonoids, of which the flavonoids are the most important. The two bestknown groups of flavonoids are the anthocyanins and condensed tannins (also called proanthocyanidins).  Anthocyanins are responsible for the red colour in grapes. The condensed tannins (proanthocyanidins) are responsible for some major wine sensorial properties (astringency, browning, and turbidity) and areinvolved in the wine ageing processes. This review summarises flavonoid synthesis in the grape berry and the impact of environmental factors on the accumulation rate during ripening of each of the flavonoids.  The impact of the accumulated flavonoids in grapes and the resulting impact on the sensorial aspects of the wine are also discussed

A 2d in vivo approach to study photosynthesis in grape berry

Is argued that fruit photosynthesis serves mainly as a respiratory CO2 refixation mechanism [1] but its contribution to growth and metabolism, localization and dynamics during fruit development are poorly known. Unlike the leaves, fruit volume imposes a constraint to photosynthesis by limiting light penetration. However, the patterns of chlorophyll distribution are apparently independent of a light intensity gradient. Microscopic observations of transversal slices of green stage grape berries (6-8 weeks after fruit set) of Alvarinho cultivar, revealed that exocarp cells, mesocarp cells next to vascular bundles, and seed coat cells present higher chlorophyll contents than inner mesocarp cells. The photosynthetic activity was determined on this material by Imaging-PAM fluorometry, a powerful tool for 2D mapping of in vivo photosynthesis. In 2 mm-thick grape berry discs, chlorophyll fluorescence parameters were estimated (Fv/Fm and II), and rapid light curves (RLC) were performed. Exocarp and seed coats of green berries showed the highest Fv/Fm values (ca. 0.6-0.7), and mesocarp cells around 85% of that value. Exocarp from mature grapes maintained Fv/Fm values during maturation, but in mesocarp and seed coats this value strongly decreased. ETRr were very sensitive to increasing light intensities and decreased with grape berry maturation. Our future prospects include the implication of photosynthesis on grape berry solute contents (sugars, acids), fruit and seed development.Fundação para a Ciência e a Tecnologia (research project no. PTDC/AGR-ALI/100636/2008

Novel, technical advance: a new grapevine transpiration prototype for grape berries and whole bunch based on relative humidity sensors

Grape berry transpiration is considered an important process during maturation, but scientific evidence is scarce. In the literature, there is only one report showing reduced maturation when bunch transpiration is artificially slowed down. Traditionally, grape berry transpiration has been measured by weighing grape berries on scale for a given time, correctly assuming that the weight reduction is due to water lost. Commercially available instruments adequate to measure gas exchange in small fruits are not suitable for whole grape berry bunch. Here, we present an open differential chamber system that can be used with isolated grape berries or alternatively with a whole grape berry bunch for measuring grape berry/bunch transpiration based on the use of relative humidity sensors from Vaisala. When used with isolated grape berries, open differential chamber system validation was made by using Tempranillo grape berries collected at different phenological stages. For the whole bunch transpiration prototype, two different validations were made. Firstly, measurements were made inserting inside the chamber an increasing number of Eppendorf tubes filled with water. Secondly, transpiration was measured in whole Tempranillo bunches sampled at different phenological stages. An important output of this work is that the fact of detaching the bunch from the plant did not change the bunch gas exchange rates at least for several hours

Anatomical aspects of grape berry development

The anatomical development of the sultana-grape berry has been followed from anthesis to maturity on material grown under glasshouse and field conditions including field-grown clonal lines differing in final fruit size. Fresh weight, volume, berry dimensions, moisture content and dry weight were measured on whole berries. Pericarp growth was studied at the cell level. Pericarp growth is basically responsible for the overall growth of the berry and this tissue represents 64% of the mature fruit's total volume. The period required for complete berry development (approximately 100 days) falls into two major growth periods separated by a lag phase. Before the lag phase pericarp growth results partly from cell division but mainly from cell enlargement. After the lag phase pericarp growth results entirely from cell enlargement. Cell division in the pericarp ceases about one week before the lag phase. Berry size differences between clonal lines were primarily due to differences in the size of pericarp cells. Berry size differences between fruits grown in the glasshouse and in the field at Merbein were due to differences in both pericarp cell number and cell size