Grape Flavonoid Evolution and Composition Under Altered Light and Temperature Conditions in Cabernet Sauvignon (Vitis vinifera L.).
ABSTRACT: The evolution of flavonoids under altered temperature and light conditions in the fruit zone was followed in Cabernet Sauvignon (Vitis vinifera L.) grapes during ripening. The study was conducted over two consecutive seasons in 2010/2011 and 2011/2012 comprising two main treatments in which the light quantity was manipulated in the bunch zone: (1) standard (STD) with no lateral shoot or leaf removal and (2) leaf removal west (LRW) treatment with leaf removal on the western side of the bunch zone. Furthermore, the light quality was altered by installing ultraviolet B-suppression sheets within the bunch zone in both seasons. Tannin evolution was dependent on the prevailing light quality/quantity and temperatures during berry development in a particular season. Grape seed tannin accumulation coincided with seed development and commenced at the early stages of berry development. Seed proanthocyanidin composition was not influenced by the treatments. The largest impact on proanthocyanidin accumulation and structure in the skin was due to seasonal variations highlighting the complex interaction between light quality and/or quantity across the two growing seasons and eventually the complex interaction with temperature. Flavonol accumulation was significantly influenced by the light quality, which is known to be the main abiotic driver of flavonol biosynthesis regulation. Anthocyanin concentration and content were largely dependent on the temperature and light quality in a particular season. Anthocyanin composition was altered by the season rather than the treatment.
Project description:Light quantity and quality modulate grapevine development and influence berry metabolic processes. Here we studied light as an information signal for developing and ripening grape berries. A Vitis vinifera Sauvignon Blanc field experiment was used to identify the impacts of UVB on core metabolic processes in the berries under both high light (HL) and low light (LL) microclimates. The primary objective was therefore to identify UVB-specific responses on berry processes and metabolites and distinguish them from those responses elicited by variations in light incidence. Canopy manipulation at the bunch zone via early leaf removal, combined with UVB-excluding acrylic sheets installed over the bunch zones resulted in four bunch microclimates: (1) HL (control); (2) LL (control); (3) HL with UVB attenuation and (4) LL with UVB attenuation. Metabolite profiles of three berry developmental stages showed predictable changes to known UV-responsive compound classes in a typical UV acclimation (versus UV damage) response. Interestingly, the berries employed carotenoids and the associated xanthophyll cycles to acclimate to UV exposure and the berry responses differed between HL and LL conditions, particularly in the developmental stages where berries are still photosynthetically active. The developmental stage of the berries was an important factor to consider in interpreting the data. The green berries responded to the different exposure and/or UVB attenuation signals with metabolites that indicate that the berries actively managed its metabolism in relation to the exposure levels, displaying metabolic plasticity in the photosynthesis-related metabolites. Core processes such as photosynthesis, photo-inhibition and acclimation were maintained by differentially modulating metabolites under the four treatments. Ripe berries also responded metabolically to the light quality and quantity, but mostly formed compounds (volatiles and polyphenols) that have direct antioxidant and/or "sunscreening" abilities. The data presented for the green berries and those for the ripe berries conform to what is known for UVB and/or light stress in young, active leaves and older, senescing tissues respectively and provide scope for further evaluation of the sink/source status of fruits in relation to photosignalling and/or stress management.
Project description:Flavonols constitute a group of flavonoids with important photoprotective roles in plants. In addition, flavonol content and composition greatly influences fruit quality. We previously demonstrated that the grapevine R2R3-MYB transcription factor (TF) VviMYBF1 promotes flavonol accumulation by inducing the expression of flavonol synthase (VviFLS1/VviFLS4), a key step of the initial flavonol pathway. Despite this, gene networks underlying flavonol modification in grapevine including both structural and regulatory genes remain poorly understood. In order to identify flavonol modifying genes and TFs acting downstream of VviMYBF1 a microarray-based transcriptome analysis was performed on grapevine hairy roots ectopically expressing VviMYBF1 or a Green Fluorescent Protein as control. VviFLS1 was induced in VviMYBF1 transgenic roots and glycosylated flavonols accumulated significantly compared with control lines. Among the differentially expressed genes, potential flavonol-modifying enzymes with predicted rhamnosyltransferase (e.g., RhaT1) or glycosyltransferase (e.g., GT3) activities were identified. In addition, important TFs of the MYB and bZIP families such as the proanthocyanidin regulator VviMYBPA1 and the UV-B light responsive HY5 homolog VviHYH were significantly altered in their expression pattern by overexpression of VviMYBF1. Co-temporal expression analysis demonstrated positive correlation of VviMYBF1 with VviFLS1, VviGT3, and VviRhaT1 during berry development and in fruits ripened with different light and UV-B radiation conditions at field. These results show that VviMYBF1 overexpression led to the identification of novel genes of the flavonol pathway and that the flavonol modifying machinery can be influenced by agricultural practices to optimize flavonol composition in grapes.
Project description:Fencing is an effective and practical method for restoring degraded grasslands in northern China. However, little is known about the role of excess litter accumulation due to long-term fencing in regulating abiotic environment and driving changes in community structure and function. We conducted a three-year field experiment in two fenced grasslands in Inner Mongolia, and monitored light quantity, soil temperature, and soil moisture continuously, and determined community height, community aboveground net primary productivity (ANPP), and the relative dominance of different plant functional groups. Litter accumulation reduced light quantity and soil temperature but increased soil moisture. The regulating effects of litter accumulation on soil temperature and soil moisture fluctuated temporally and gradually weakened over the growing season. Litter accumulation also altered community vertical structure and function by increasing community height and ANPP. The increase in soil moisture increased the relative dominance of rhizome grasses but suppressed bunch grasses, thereby shifting bunch grass grasslands to rhizome grass grasslands. Our findings provide a potential mechanism for community succession in the context of litter accumulation in fenced grasslands and indicate that the vegetation and ecosystem services of degraded grasslands are improved after appropriate fencing.
Project description:Viticulture is widely practiced in dry regions, where the grapevine is greatly exposed to water stress. Optimizing plant water use efficiency (WUE) without affecting crop yield, grape and wine quality is crucial to limiting use of water for irrigation and to significantly improving viticulture sustainability. This study examines the use in vineyards of particle film technology (engineered kaolin) and compares it to a film-forming antitranspirant (pinolene), traditionally used to limit leaf water loss, and to an untreated control. The trial was carried out under field conditions over three growing seasons, during which moderate to very severe plant water stress (down to -1.9 MPa) was measured through stem water potential. Leaf stomatal conductance (gs) and photosynthesis rate (An) were measured during the seasons and used to compute intrinsic WUE (WUEi, defined as An/gs ratio). Leaf temperature was also recorded and compared between treatments. Bunch quantity, bunch and berry weight, sugar accumulation, anthocyanin and flavonoid contents were measured. Finally, microvinifications were performed and resultant wines subjected to sensory evaluation.Results showed that the use of kaolin increased grapevine intrinsic WUE (+18% on average as compared to unsprayed vines) without affecting berry and bunch weight and quantity, or sugar level. Anthocyanin content increased (+35%) in kaolin treatment, and the wine was judged more attractive (p-value <0.05) and slightly more appreciated (p-value < 0.1) than control. Pinolene did not increase WUEi, limiting An more than gs; grapes with this treatment contained lower sugar and anthocyanin content than control, and the obtained wine was the least appreciated. This study demonstrates that particle film technology can improve vine WUEi and wine quality at the same time, while traditional antitranspirants were not as effective for these purposes. This positive effect can be used in interaction with other already-demonstrated uses of particle film technology, such as pest control and sunburn reduction, in order to achieve more sustainable vineyard management.
Project description:Grapevine red blotch disease (GRBD) is a recently identified viral disease that affects grapevines. GRBD has been shown to impact grapevine physiology and grape composition by altering specific ripening events. However, no studies have been reported on the impact of GRBD on wine composition and its sensory attributes. This study evaluated the impact of GRBD on wine primary and secondary metabolites, in addition to its sensory properties, when making wines from Cabernet Sauvignon and Merlot grapes during two seasons. Wines made with GRBD-impacted fruit were lower in ethanol content when compared to wines made with grapes from healthy grapevines. This was attributed to the lower total soluble sugar (TSS) levels of diseased grapes due to delayed ripening at harvest. GRBD impacted wine phenolic composition by decreasing anthocyanin concentrations and increasing flavonol concentrations in some instances. Additionally, proanthocyanidin concentrations were also consistently higher in GRBD wines compared to wines made from healthy fruit. Descriptive analysis demonstrated that GRBD can impact wine style by altering aroma, flavor, and mouthfeel attributes. However, the extent of GRBD impact on wine composition and sensory properties were site and season dependent.
Project description:Flavonols constitute a group of flavonoids with important photoprotective roles in plants. In addition, flavonol content and composition greatly influences fruit quality. We previously demonstrated that the grapevine R2R3-MYB transcription factor VviMYBF1 promotes flavonol accumulation by inducing the expression of flavonol synthase (VviFLS1/VviFLS4), a key step of the initial flavonol pathway. Despite this, gene networks underlying flavonol modification in grapevine including both structural and regulatory genes remain poorly understood. In order to identify flavonol modifying genes and transcription factors acting downstream of VviMYBF1 a microarray-based transcriptome analysis was performed on grapevine hairy roots ectopically expressing VviMYBF1 or a Green Fluorescent Protein as control. VviFLS1 was induced in VviMYBF1 transgenic roots and flavonols, mainly quercetin 3-glucoside and quercetin 3-galactoside, accumulated significantly compared with control lines. Among the differentially expressed genes, potential flavonol-modifying enzymes with predicted rhamnosyltransferase (e.g. RhaT1) or glycosyltransferase (e.g. GT3) activities were identified. In addition, important transcription factors of the MYB and bZIP families such as the proanthocyanidin regulator VviMYBPA1 and the UV-B light responsive HY5 homologue VviHYH were significantly altered in their expression pattern by overexpression of VviMYBF1. In silico promoter sequence analysis confirmed that the significantly differentially expressed genes contained binding sites similar to known MYB recognition elements. Co-temporal expression analysis demonstrated positive correlation of VviMYBF1 and accumulation of glycosylated flavonols with VviGT3 and VviRhaT1 during berry development and in fruits ripened with different light and UV-B radiation conditions at field. These results show that VviMYBF1 overexpression led to the identification of novel genes of the flavonol pathway and that the flavonol modifying machinery can be influenced by agricultural practices to optimize flavonol composition in grapes. Overall design: Transgenic hairy roots were generated by infiltration of Vitis vinifera cv. ‘Chardonnay’ leaves with Agrobacterium rhizogenes carrying either VviMYBF1 or GFP (control) cDNA. To analyze differential gene expression between VviMYBF1 and GFP control hairy roots, total RNA was extracted from stable transgenic lines and subsequent array hybridization and microarray analysis was performed using the custom Affymetrix GrapeGen chip.
Project description:Under the double cropping system, berries usually showed significant quality variations in the summer and winter seasons. In the two-year/four-consecutive-season study, two table grapes of 'Muscat Hamburg' and 'Victoria' were investigated to determine the phenolic compounds in their berries. Different from those of the summer season, the berries in the winter season suffered no high-temperature stress since veraison to harvest in 2014 and 2015. The variations in the season temperatures led to a higher anthocyanin concentration in the winter season berries of 'Muscat Hamburg' grapes than that in the summer berries, while the summer season berries had higher proportions of acylated and methylated anthocyanins than those in the winter season berries. Similar to the anthocyanins, the winter season berries also had a higher flavonol concentration in both varieties. Transcriptome analysis showed that the upregulated genes involved in the flavonoid pathway in the winter season berries were agreed with the changes found in the metabolites. However, the influence of the growing seasons on the flavanols was not consistent in the two varieties, and the variations in <i>VviLARs</i> between the grapes of 'Muscat Hamburg' and 'Victoria' might be the cause. This research helped us better understand the double cropping system and how the climate factors affected the phenolic compounds in the double cropping system.
Project description:Light conditions significantly influence grape berry ripening and the accumulation of phenolic compounds, but the underlying molecular basis remains partially understood. Here, we applied integrated transcriptomics and pathway-level metabolomics analyses to investigate the effect of cluster bagging during various developmental stages on phenolic metabolism in Cabernet Sauvignon grapes. Bagging treatments had limited effects on berry quality attributes at harvest and did not consistently affect phenolic acid biosynthesis between seasons. Significantly elevated flavan-3-ol and flavonol contents were detected in re-exposed berries after bagging during early-developmental stages, while bagging after véraison markedly inhibited skin anthocyanin accumulation. Several anthocyanin derivatives and flavonol glycosides were identified as marker phenolic metabolites for distinguishing bagged and non-bagged grapes. Coordinated transcriptional changes in the light signaling components CRY2 and HY5/HYHs, transcription regulator MYBA1, and enzymes LAR, ANR, UFGT and FLS4, coincided well with light-responsive biosynthesis of the corresponding flavonoids. The activation of multiple hormone signaling pathways after both light exclusion and re-exposure treatments was inconsistent with the changes in phenolic accumulation, indicating a limited role of plant hormones in mediating light/darkness-regulated phenolic biosynthesis processes. Furthermore, gene-gene and gene-metabolite network analyses discovered that the light-responsive expression of genes encoding bHLH, MYB, WRKY, NAC, and MADS-box transcription factors, and proteins involved in genetic information processing and epigenetic regulation such as nucleosome assembly and histone acetylation, showed a high positive correlation with grape berry phenolic accumulation in response to different light regimes. Altogether, our findings provide novel insights into the understanding of berry phenolic biosynthesis under light/darkness and practical guidance for improving grape features. Overall design: Grape Berries from T1 treatment (CBFH) at E-L 31, 35, 36, 37, and 38 stages, T8 treatment (CBV) at E-L 36, 37, and 38 stages, and the control group at E-L 29, 31, 35, 36, 37, and 38 stages during the 2012 growing season were selected to conduct the transcriptome profiling analysis based on the physiological and metabolic results. Each sample contain three biological replicates.
Project description:Citrus fruits exhibit a high level of different phytoconstituents, of which the changes in the different parts of the fruit during ripening have not been thoroughly studied yet. Thus, in this study, we have investigated how different parts of pomelo fruit (<i>Citrus grandis</i> L.) are modified throughout the development of two consecutive growing seasons. In detail, the main phytochemical compounds, such as total phenolic content, total flavonoid content, antioxidant capacity, DPPH free radical scavenging activity, Ferric reducing antioxidant power (FRAP), and naringin and tannin content, were analyzed. A systematic metabolism of these compounds was found during the development of the fruit, but some pomelo tissues showed a fluctuating trend, suggesting a dependence on the different growing season. Focusing on the tissue distribution of these compounds, the fruit membrane contained the highest level of total phenolic and flavonoid content; fruit flavedo displayed the highest antioxidant capacities and FRAP activities, whereas maximum accumulation of naringin was noticed in fruit albedo. Instead, the highest DPPH free radical scavenging activity and tannin contents were found in the pomelo juice. Regarding the distribution of compounds, a possible bias pattern for the accumulation of those compounds has been noticed throughout the fruit development. From the GC-MS analysis, a total of 111 compounds were identified, where 91 compounds were common in both seasons. Overall, these results could be useful for the food processing industry as guidelines for excellent quality foods and for introducing health-beneficial products and components into our daily diets.
Project description:Although biochemists and geneticists have studied the cotton flower for more than one century, little is known about the molecular mechanisms underlying the dramatic color change that occurs during its short developmental life following blooming. Through the analysis of world cotton germplasms, we found that all of the flowers underwent color changes post-anthesis, but there is a diverse array of petal colors among cotton species, with cream, yellow and red colors dominating the color scheme. Genetic and biochemical analyses indicated that both the original cream and red colors and the color changes post-anthesis were related to flavonoid content. The anthocyanin content and the expression of biosynthesis genes were both increased from blooming to one day post-anthesis (DPA) when the flower was withering and undergoing abscission. Our results indicated that the color changes and flavonoid biosynthesis of cotton flowers were precisely controlled and genetically regulated. In addition, flavonol synthase (FLS) genes involved in flavonol biosynthesis showed specific expression at 11 am when the flowers were fully opened. The anthocyanidin reductase (ANR) genes, which are responsible for proanthocyanidins biosynthesis, showed the highest expression at 6 pm on 0 DPA, when the flowers were withered. Light showed primary, moderate and little effects on flavonol, anthocyanin and proanthocyanidin biosynthesis, respectively. Flavonol biosynthesis was in response to light exposure, while anthocyanin biosynthesis was involved in flower color changes. Further expression analysis of flavonoid genes in flowers of wild type and a flavanone 3-hydroxylase (F3H) silenced line showed that the development of cotton flower color was controlled by a complex interaction between genes and light. These results present novel information regarding flavonoids metabolism and flower development.