Dataset Information

Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.

ABSTRACT: BACKGROUND:Structural genes of the phenyl-propanoid pathway which encode flavonoid 3'- and 3',5'-hydroxylases (F3'H and F3'5'H) have long been invoked to explain the biosynthesis of cyanidin- and delphinidin-based anthocyanin pigments in the so-called red cultivars of grapevine. The relative proportion of the two types of anthocyanins is largely under genetic control and determines the colour variation among red/purple/blue berry grape varieties and their corresponding wines. RESULTS:Gene fragments of VvF3'H and VvF3'5'H, that were isolated from Vitis vinifera 'Cabernet Sauvignon' using degenerate primers designed on plant homologous genes, translated into 313 and 239 amino acid protein fragments, respectively, with up to 76% and 82% identity to plant CYP75 cytochrome P450 monooxygenases. Putative function was assigned on the basis of sequence homology, expression profiling and its correlation with metabolite accumulation at ten different ripening stages. At the onset of colour transition, transcriptional induction of VvF3'H and VvF3'5'H was temporally coordinated with the beginning of anthocyanin biosynthesis, the expression being 2-fold and 50-fold higher, respectively, in red berries versus green berries. The peak of VvF3'5'H expression was observed two weeks later concomitantly with the increase of the ratio of delphinidin-/cyanidin-derivatives. The analysis of structural genomics revealed that two copies of VvF3'H are physically linked on linkage group no. 17 and several copies of VvF3'5'H are tightly clustered and embedded into a segmental duplication on linkage group no. 6, unveiling a high complexity when compared to other plant flavonoid hydroxylase genes known so far, mostly in ornamentals. CONCLUSION:We have shown that genes encoding flavonoid 3'- and 3',5'-hydroxylases are expressed in any tissues of the grape plant that accumulate flavonoids and, particularly, in skin of ripening red berries that synthesise mostly anthocyanins. The correlation between transcript profiles and the kinetics of accumulation of red/cyanidin- and blue/delphinidin-based anthocyanins indicated that VvF3'H and VvF3'5'H expression is consistent with the chromatic evolution of ripening bunches. Local physical maps constructed around the VvF3'H and VvF3'5'H loci should help facilitate the identification of the regulatory elements of each isoform and the future manipulation of grapevine and wine colour through agronomical, environmental and biotechnological tools.

SUBMITTER: Castellarin SD

PROVIDER: S-EPMC1403756 | biostudies-literature | 2006 Jan

REPOSITORIES: biostudies-literature

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Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.

Castellarin Simone D SD Di Gaspero Gabriele G Marconi Raffaella R Nonis Alberto A Peterlunger Enrico E Paillard Sophie S Adam-Blondon Anne-Francoise AF Testolin Raffaele R

BMC genomics 20060124

<h4>Background</h4>Structural genes of the phenyl-propanoid pathway which encode flavonoid 3'- and 3',5'-hydroxylases (F3'H and F3'5'H) have long been invoked to explain the biosynthesis of cyanidin- and delphinidin-based anthocyanin pigments in the so-called red cultivars of grapevine. The relative proportion of the two types of anthocyanins is largely under genetic control and determines the colour variation among red/purple/blue berry grape varieties and their corresponding wines.<h4>Results< ...[more]

PMID: 16433923

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Project description:BACKGROUND: Symptoms of grapevine leafroll disease (GLRD) in red-fruited wine grape (Vitis vinifera L.) cultivars consist of green veins and red and reddish-purple discoloration of inter-veinal areas of leaves. The reddish-purple color of symptomatic leaves may be due to the accumulation of anthocyanins and could reflect an up-regulation of genes involved in their biosynthesis. RESULTS: We examined six putative constitutively expressed genes, Ubiquitin, Actin, GAPDH, EF1-a, SAND and NAD5, for their potential as references for normalization of gene expression in reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Using the geNorm program, a combination of two genes (Actin and NAD5) was identified as the stable set of reference genes for normalization of gene expression data obtained from grapevine leaves. By using gene-specific RT-qPCR in combination with a reliable normalization factor, we compared relative expression of the flavonoid biosynthetic pathway genes between leaves infected with Grapevine leafroll-associated virus 3 (GLRaV-3) and exhibiting GLRD symptoms and virus-free green leaves obtained from a red-fruited wine grape cultivar (cv. Merlot). The expression levels of these different genes ranged from two- to fifty-fold increase in virus-infected leaves. Among them, CHS3, F3'5'H, F3H1, LDOX, LAR1 and MybA1 showed greater than 10-fold increase suggesting that they were expressed at significantly higher levels in virus-infected symptomatic leaves. HPLC profiling of anthocyanins extracted from leaves indicated the presence of cyanidin-3-glucoside and malvidin-3-glucoside only in virus-infected symptomatic leaves. The results also showed 24% higher levels of flavonols in virus-infected symptomatic leaves than in virus-free green leaves, with quercetin followed by myricetin being the predominant compounds. Proanthocyanidins, estimated as total tannins by protein precipitation method, were 36% higher in virus-infected symptomatic leaves when compared to virus-free green leaves. CONCLUSIONS: The results, the first example to our knowledge, showed that modulation of the flavonoid biosynthetic pathway occurred in GLRaV-3-infected leaves of a red-fruited wine grape cultivar (cv. Merlot) leading to de novo synthesis of two classes of anthocyanins. These anthocyanins have contributed to the expression of reddish-purple color of virus-infected grapevine leaves exhibiting GLRD symptoms.

Project description:Although deficit irrigation is used to improve fruit quality in healthy grapevines, it can potentially amplify negative effects of viral disease and reduce fruit quality in Grapevine Red Blotch Virus (GRBV) infected grapevines. Therefore, a 2-year field experiment was conducted to understand the interaction between GRBV infection and water deficits on disease development and vine physiology. Well-watered (WW) vines were irrigated at 100% of estimated crop evapotranspiration (ETc), while water deficit (WD) vines received water at 66 and 50% ETc in 2017 and 2018, respectively. Healthy (GRBV-) and infected (GRBV+) vines were confirmed by PCR assays. There were no significant effects of water deficits on foliar symptom onset in either year, but more severe water deficits in 2018 resulted in a more rapid symptom progression. GRBV+ vines had a higher ?stem compared to GRBV- vines, but the effects of virus only appeared post-veraison and corresponded to decreased leaf gas exchange. In general, vine vegetative and reproductive growth were not reduced in GRBV+ vines. Yields were highest in WW/GRBV+ vines due to larger clusters containing larger berries. Consistent treatment effects on berry primary chemistry were limited to sugars, with no interactions between factors. Water deficits were able to somewhat increase berry anthocyanin concentration in GRBV+ fruit, but the effects were dependent on year. By comparison, virus status and water deficits interacted on skin tannins concentration such that they were decreased in WD/GRBV+ vines, but increased in WD/GRBV- vines. Water deficits had no effect on seed phenolics, with only virus status having a significant diminution. Although keeping GRBV+ vines well-watered may mitigate some of the negative effects of GRBD, these results suggest that water deficits will not improve overall fruit quality in GRBV+ vines. Ultimately, the control of fruit ripening imparted by GRBV infection seems to be stronger than abiotic control imparted by water deficits.

Project description:Background:Anthocyanin compounds playing multiple biological functions can be synthesized in different parts of barley (Hordeum vulgare L.) plant. The diversity of anthocyanin molecules is related with branching the pathway to alternative ways in which dihydroflavonols may be modified either with the help of flavonoid 3'-hydroxylase (F3'H) or flavonoid 3',5'-hydroxylase (F3'5'H)-the cytochrome P450-dependent monooxygenases. The F3'H and F3'5'H gene families are among the least studied anthocyanin biosynthesis structural genes in barley. The aim of this study was to identify and characterise duplicated copies of the F3'H and F3'5'H genes in the barley genome. Results:Four copies of the F3'5'H gene (on chromosomes 4HL, 6HL, 6HS and 7HS) and two copies of the F3'H gene (on chromosomes 1HL and 6HS) were identified in barley genome. These copies have either one or two introns. Amino acid sequences analysis demonstrated the presence of the flavonoid hydroxylase-featured conserved motifs in all copies of the F3'H and F3'5'H genes with the exception of F3'5'H-3 carrying a loss-of-function mutation in a conservative cytochrome P450 domain. It was shown that the divergence between F3'H and F3'5'H genes occurred 129 million years ago (MYA) before the emergence of monocot and dicot plant species. The F3'H copy approximately occurred 80 MYA; the appearance of F3'5'H copies occurred 8, 36 and 91 MYA. qRT-PCR analysis revealed the tissue-specific activity for some copies of the studied genes. The F3'H-1 gene was transcribed in aleurone layer, lemma and pericarp (with an increased level in the coloured pericarp), whereas the F3'H-2 gene was expressed in stems only. The F3'5'H-1 gene was expressed only in the aleurone layer, and in a coloured aleurone its expression was 30-fold higher. The transcriptional activity of F3'5'H-2 was detected in different tissues with significantly higher level in uncoloured genotype in contrast to coloured ones. The F3'5'H-3 gene expressed neither in stems nor in aleurone layer, lemma and pericarp. The F3'5'H-4 gene copy was weakly expressed in all tissues analysed. Conclusion:F3'H and F3'5'H-coding genes involved in anthocyanin synthesis in H. vulgare were identified and characterised, from which the copies designated F3'H-1, F3'H-2, F3'5'H-1 and F3'5'H-2 demonstrated tissue-specific expression patterns. Information on these modulators of the anthocyanin biosynthesis pathway can be used in future for manipulation with synthesis of diverse anthocyanin compounds in different parts of barley plant. Finding both the copies with tissue-specific expression and a copy undergoing pseudogenization demonstrated rapid evolutionary events tightly related with functional specialization of the duplicated members of the cytochrome P450-dependent monooxygenases gene families.

Project description:Monastrell grapevines grafted on the rootstocks 140Ru, 1103P, 41B, 110R, and 161-49C were subjected to regulated deficit irrigation (RDI) and partial root-zone irrigation (PRI). We analyzed the effects of the rootstock and irrigation method on the phenolic concentration in different berry tissues, its dilution/concentration due to the berry size, the anatomical and morphological traits of berries related to the phenolic compounds concentration, and the relationships of all these parameters with the final berry and wine phenolic content. The rootstock had an important effect on the accumulation of total phenolic compounds and anthocyanins in the skin (berries from 110R and 140Ru had the highest values). Moreover, the rootstock modified some anatomical and morphological characteristics that had a direct relationship with the final phenolic compounds concentration in the must. Large grapes and high must percentages (110R and 140Ru) produced a dilution effect, whereas small berries and a low must percentage increased the concentration (161-49C). For 110R, the small size of the cells of the epidermis and hypodermis in the grapes also could have contributed to the high phenolic compounds concentration in the skin. The percentage of cells in the skin with a uniform coloration was positively correlated with its total phenolic compounds and anthocyanins concentration and also with the phenolic quality of the wine. The PRI modified some specific morphological/anatomical skin/berry traits, and these may have contributed to important changes in the final concentration of phenolic compounds, depending on the rootstock. The better phenolic quality of the must and wines observed in some rootstocks under PRI could be due to smaller cells in the epidermis and hypodermis of the skin (161-49C), a higher percentage of cells with a uniform coloration in the hypodermis (110R), or a lower number of seeds per berry (161-49C). In contrast, the lower phenolic compounds concentration in the must of grapes observed in the most vigorous rootstocks under PRI could be due to a greater thickness of the epidermis (140Ru), greater cuticle thickness (41B), a higher number of seeds (140Ru), a lower skin/pulp ratio and percentage of skin (140Ru), a greater percentage of cells in the epidermis without coloration or with large inclusions, and a lower percentage of cells with a uniform coloration in the epidermis (140Ru). The final quality of the grape is related to some changes in histological and morphological aspects of the grape produced by the rootstock and irrigation strategy.

Dataset Information

Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.

Publications

Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.

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