Instability of anthocyanin accumulation in Vitis vinifera L. var. Gamay Freaux suspension cultures

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinitera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of 2.73 +/- 0.15, 1.45 +/- 0.04, 0.77 +/- 0.024 and 0.27 +/- 0.04 CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and 84% for V vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be 9.7%, ranging from 4 to 17%. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line, showed greater potential in enhanced the anthocyanin production.

Genetics of berry colour and anthocyanin content variation in grapevine (Vitis vinifera L.subsp. vinifera)

Dissertation presented in fulfillment of the requirements for the Degree of Doctor of Philosophy in Biology (Molecular Genetics) at the Instituto de Tecnologia Química e Biol��gica da Universidade Nova de Lisboa

Factors affecting distribution and mobility of trace elements (Cu, Pb, Zn) in a perennial grapevine (Vitis vinifera L.) in the Champagne region of France

Soil and Vitis vinifera L (coarse and fine roots, leaves, berries) concentration and geochemical partitioning of Cu, Pb and Zn were determined in a contaminated calcareous Champagne plot to assess their mobility and transfer. Accumulation ratios in roots remained low (0.1-0.4 for Cu and Zn, <0.05 for Pb). Differences between elements resulted from vegetation uptake strategy and soil partitioning. Copper, significantly associated with the oxidisable fraction (27.8%), and Zn with the acid soluble fraction (33.3%), could be mobilised by rhizosphere acidification and oxidisation, unlike Pb, essentially contained in the reducible fraction (72.4%). Roots should not be considered as a whole since the more reactive fine roots showed higher accumulation ratios than coarse ones. More sensitive response of fine roots, lack of correlation between chemical extraction results and vegetation concentrations, and very limited translocation to aerial parts showed that fine root concentrations should be used when assessing bioavailability. (C) 2008 Elsevier Ltd. All rights reserved.

Phenolic profile of Sercial and Tinta Negra Vitis vinifera L. grape skins by HPLC–DAD–ESI-MSn: novel phenolic compounds in Vitis vinifera L. grape

This study represents the first phytochemical research of phenolic components of Sercial and Tinta Negra Vitis vinifera L. The phenolic profiles of Sercial and Tinta Negra V. vinifera L. grape skins (white and red varieties, respectively) were established using high performance liquid chromatography–diode array detection–electrospray ionisation tandem mass spectrometry (HPLC–DAD–ESI-MSn), at different ripening stages (véraison and maturity). A total of 40 phenolic compounds were identified, which included 3 hydroxybenzoic acids, 8 hydroxycinnamic acids, 4 flavanols, 5 flavanones, 8 flavonols, 4 stilbenes, and 8 anthocyanins. For the white variety, in both ripening stages, hydroxycinnamic acids and flavonols were the main phenolic classes, representing about 80% of the phenolic composition. For red variety, at véraison, hydroxycinnamic acids and flavonols were also the predominant classes (71%), but at maturity, anthocyanins represented 84% of the phenolic composition. As far as we know, 10 compounds were reported for the first time in V. vinifera L. grapes, namely protocatechuic acid-glucoside, p-hydroxybenzoyl glucoside, caftaric acid vanilloyl pentoside, p-coumaric acid-erythroside, naringenin hexose derivate, eriodictyol-glucoside, taxifolin-pentoside, quercetin-glucuronide-glucoside, malylated kaempferol-glucoside, and resveratrol dimer. These novel V. vinifera L. grape components were identified based on their MSn fragmentation profile. This data represents valuable information that may be useful to oenological management and to valorise these varieties as sources of bioactive compounds.

Solid phase microextraction as a reliable alternative to conventional extraction techniques to evaluate the pattern of hydrolytically released components in Vitis vinifera L. grapes

In present research, headspace solid-phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC–qMS), was evaluated as a reliable and improved alternative to the commonly used liquid–liquid extraction (LLE) technique for the establishment of the pattern of hydrolytically released components of 7 Vitis vinifera L. grape varieties, commonly used to produce the world-famous Madeira wine. Since there is no data available on their glycosidic fractions, at a first step, two hydrolyse procedures, acid and enzymatic, were carried out using Boal grapes as matrix. Several parameters susceptible of influencing the hydrolytic process were studied. The best results, expressed as GC peak area, number of identified components and reproducibility, were obtained using ProZym M with b-glucosidase activity at 35 °C for 42 h. For the extraction of hydrolytically released components, HS-SPME technique was evaluated as a reliable and improved alternative to the conventional extraction technique, LLE (ethyl acetate). HS-SPME using DVB/CAR/PDMS as coating fiber displayed an extraction capacity two fold higher than LLE (ethyl acetate). The hydrolyzed fraction was mainly characterized by the occurrence of aliphatic and aromatic alcohols, followed by acids, esters, carbonyl compounds, terpenoids, and volatile phenols. Concerning to terpenoids its contribution to the total hydrolyzed fraction is highest for Malvasia Cândida (23%) and Malvasia Roxa (13%), and their presence according previous studies, even at low concentration, is important from a sensorial point of view (can impart floral notes to the wines), due to their low odor threshold (μg/L). According to the obtained data by principal component analysis (PCA), the sensorial properties of Madeira wines produced by Malvasia Cândida and Malvasia Roxa could be improved by hydrolysis procedure, since their hydrolyzed fraction is mainly characterized by terpenoids (e.g. linalool, geraniol) which are responsible for floral notes. Bual and Sercial grapes are characterized by aromatic alcohols (e.g. benzyl alcohol, 2-phenylethyl alcohol), so an improvement in sensorial characteristics (citrus, sweet and floral odors) of the corresponding wines, as result of hydrolytic process, is expected.

Optimisation of solid-phase microextraction combined with gas chromatography–mass spectrometry based methodology to establish the global volatile signature in pulp and skin of Vitis vinifera L. grape varieties

The volatiles (VOCs) and semi-volatile organic compounds (SVOCs) responsible for aroma are mainly present in skin of grape varieties. Thus, the present investigation is directed towards the optimisation of a solvent free methodology based on headspace-solid-phase microextraction (HS-SPME) combined with gas chromatography–quadrupole mass spectrometry (GC–qMS) in order to establish the global volatile composition in pulp and skin of Bual and Bastardo Vitis vinifera L. varieties. A deep study on the extraction-influencing parameters was performed, and the best results, expressed as GC peak area, number of identified compounds and reproducibility, were obtained using 4 g of sample homogenised in 5 mL of ultra-pure Milli-Q water in a 20 mL glass vial with addition of 2 g of sodium chloride (NaCl). A divinylbenzene/carboxen/polydimethylsiloxane fibre was selected for extraction at 60 °C for 45 min under continuous stirring at 800 rpm. More than 100 VOCs and SVOCs, including 27 monoterpenoids, 27 sesquiterpenoids, 21 carbonyl compounds, 17 alcohols (from which 2 aromatics), 10 C13 norisoprenoids and 5 acids were identified. The results showed that, for both grape varieties, the levels and number of volatiles in skin were considerably higher than those observed in pulp. According to the data obtained by principal component analysis (PCA), the establishment of the global volatile signature of grape and the relationship between different part of grapes—pulp and skin, may be an useful tool to winemaker decision to define the vinification procedures that improves the organoleptic characteristics of the corresponding wines and consequently contributed to an economic valorization and consumer acceptance.

EFEITO do ETHEPHON em VIDEIRA 'RUBI' (Vitis vinifera L.), CULTIVADA NA REGIÃO NOROESTE do ESTADO DE SÃO PAULO

O experimento foi conduzido em vinhedo comercial da cv. Rubi, localizado na região Noroeste do Estado de São Paulo. Teve, como objetivo, verificar o efeito do ethephon, aplicado antes da poda de produção, na porcentagem de desfolhamento, tempo de realização da poda, brotação dos ramos, produção da videira, tentando reduzir os custos, melhorando a produtividade e a qualidade. Concluiu-se que a aplicação de 7500ppm de ethephon, 20 dias antes da poda, foi o tratamento que proporcionou maior porcentagem de desfolhamento, menor tempo de realização da poda, maior porcentagem de gemas brotadas e maior produção.

Níveis e épocas de aplicação de etefon sobre a coloração e a qualidade dos frutos da uva rubi (Vitis vinifera L.), cultivada na região noroeste do Estado de São Paulo

Pós-graduação em Agronomia (Horticultura) - FCA

Fenologia e efeito da desfolha parcial na produção extemporânea de efeito da desfolha parcial na produção extemporânea da videira, cultivar Cabernet Franc (Vitis vinifera L.), na região de São Roque - SP

Pós-graduação em Agronomia (Horticultura) - FCA

Fenologia e efeito da desfolha parcial na produção extemporânea de efeito da desfolha parcial na produção extemporânea da videira, cultivar Cabernet Franc (Vitis vinifera L.), na região de São Roque - SP

Pós-graduação em Agronomia (Horticultura) - FCA

Analisi del trasferimento di composti fenolici in campioni non stabilizzati di succo d?uva (vitis vinifera l.) biologico mediante tecniche di macerazione alternative

Il seguente elaborato prende le mosse da un progetto, denominato Nutrid’Uva, promosso e attivamente concretizzato dall’azienda bresciana “Cascina Belmonte” di concerto con il laboratorio del Campus di Scienze Alimentari di Cesena (Università di Bologna). In particolare, si è voluta sperimentare l���applicazione di processi innovativi per la produzione di succo d’uva bio e bevande a base di succo d’uva bio proveniente da uve biologiche coltivate nell’areale della Valténesi (sponda occidentale del Garda), stabilizzati mediante l���uso di alte pressioni idrostatiche e senza trattamento termico. In questi termini, il presente lavoro ha l���obiettivo di: delineare il quadro generale in cui si inserisce il progetto, arricchendolo con spunti tratti dalle pratiche di processo seguite in loco; determinare il contenuto delle classi polifenoliche presenti nei campioni di alcuni succhi sottoposti a differenti tecnologie di macerazione in confronto all’abituale processo di lavorazione aziendale (vitigni ibridi diretti produttori, Barbera, Merlot, Cabernet). Si è proceduto dunque attraverso l���analisi spettrofotometrica delle principali classi di composti fenolici e dell’attività antiossidante.

Nitrógeno prontamente asimilable : efecto sobre la velocidad de fermentación del jugo de uva (Vitis vinifera L.)

El propósito de este trabajo fue evaluar la incidencia de la concentración de nitrógeno prontamente asimilable (NPA) sobre la velocidad y duración de la fermentación alcohólica de los mostos de uva. El experimento se diseñó con tres tratamientos (A=testigo; B=agregado de PO4H(NH4)2 50 mg/L; C=agregado de PO4H(NH4)2 100 mg/L) y cuatro repeticiones. Se realizaron microvinificaciones con jugo de uva pasteurizado var. Chardonnay, inoculado con Saccharomyces cerevisiae cepa FCA 32. La fermentación se condujo a 25 °C. La concentración de NPA fue medida por titulación en medio formol. La velocidad de fermentación fue determinada por pérdida de peso. La velocidad máxima de fermentación se alcanzó al tercer día. Existen diferencias significativas entre la velocidad máxima alcanzada por el testigo y por los tratamientos B y C pero no hay diferencias significativas entre las velocidades máximas alcanzadas por los tratamientos B y C. La velocidad máxima de fermentación alcanzada por el tratamiento B (agregado de 50 mg/L de PO4H(NH4)2) fue 57 % superior respecto del testigo, mientras que el tratamiento C (agregado de 100 mg/L de PO4H(NH4)2) fue 53 % superior respecto del mismo testigo La velocidad máxima de fermentación aumentó con la adición de nitrógeno, pero no se observan diferencias entre las distintas dosis empleadas. La duración media de la fermentación resulta significativamente diferente para los tres tratamientos: 9.25 días para el testigo, 7.5 días para el tratamiento B y 6.25 días para el tratamiento C. El agregado de PO4H(NH4)2 disminuye la duración de la fermentación en las condiciones de trabajo. La duración de la fermentación del tratamiento B (agregado de 50 mg/L de PO4H(NH4)2) fue del 81 % respecto del testigo 100 %, mientras que el tratamiento C (agregado de 100 mg/L de PO4H(NH4)2) fue del 67 %, respecto del mismo testigo.