Pectic-β(1,4)-galactan,extensin and arabinogalactan–protein epitopes differentiate ripening stages in wine and table grape cell walls

Background and Aims

Cell wall changes in ripening grapes (Vitis vinifera) have been shown to involve re-modelling of pectin, xyloglucan and cellulose networks. Newer experimental techniques, such as molecular probes specific for cell wall epitopes, have yet to be extensively used in grape studies. Limited general information is available on the cell wall properties that contribute to texture differences between wine and table grapes. This study evaluates whether profiling tools can detect cell wall changes in ripening grapes from commercial vineyards.

Methods

Standard sugar analysis and infra-red spectroscopy were used to examine the ripening stages (green, véraison and ripe) in grapes collected from Cabernet Sauvignon and Crimson Seedless vineyards. Comprehensive microarray polymer profiling (CoMPP) analysis was performed on cyclohexanediaminetetraacetic acid (CDTA) and NaOH extracts of alcohol-insoluble residue sourced from each stage using sets of cell wall probes (mAbs and CBMs), and the datasets were analysed using multivariate software.

Key Results

The datasets obtained confirmed previous studies on cell wall changes known to occur during grape ripening. Probes for homogalacturonan (e.g. LM19) were enriched in the CDTA fractions of Crimson Seedless relative to Cabernet Sauvignon grapes. Probes for pectic-β-(1,4)-galactan (mAb LM5), extensin (mAb LM1) and arabinogalactan proteins (AGPs, mAb LM2) were strongly correlated with ripening. From green stage to véraison, a progressive reduction in pectic-β-(1,4)-galactan epitopes, present in both pectin-rich (CDTA) and hemicellulose-rich (NaOH) polymers, was observed. Ripening changes in AGP and extensin epitope abundance also were found during and after véraison.

Conclusions

Combinations of cell wall probes are able to define distinct ripening phases in grapes. Pectic-β-(1,4)-galactan epitopes decreased in abundance from green stage to véraison berries. From véraison there was an increase in abundance of significant extensin and AGP epitopes, which correlates with cell expansion events. This study provides new ripening biomarkers and changes that can be placed in the context of grape berry development.     

A disposable Laccase–Tyrosinase based biosensor for amperometric detection of phenolic compounds in must and wine

An amperometric biosensor for the detection of polyphenols in wine has been developed immobilizing the two enzymes Tyrosinase and Laccase on graphite screen printed electrodes modified with ferrocene. Different immobilization procedures have been carried out, the sensor operational parameters have been optimized, determining the best conditions and the analytical method for the analysis of samples. The biosensor has been then tested with real samples, using wines and musts supplied by Astra, experimental winery, in Imola (Italy). The biosensor gave good results when employed for wine analysis, showing a good agreement with the spectrophotometric data obtained with the Folin-Ciocalteu test, the official method for polyphenols’ analysis in wine. On the other hand, the measurements on musts and wines recently bottled, were seriously affected by the presence of an high level of free sulphur dioxide. SO₂ is the likely responsible for enzyme activity inhibition on the sensor. Further studies are currently proceeding to find out the most suitable conditions to obtain results not influenced by the presence of sulphur dioxide.     

Indigenous yeast communities in the environment of “Rovello bianco” grape variety and their use in commercial white wine fermentation

The indigenous yeast communities associated with several vineyard habitats were analysed. Wild yeasts were isolated, differentiated at strain level and identified. A phylogenetic tree based on partial 26S rRNA genes was constructed. The strains were characterized and the indigenous Saccharomyces cerevisiae GR1 was then used to carry out a vinification process and compared with a commercial yeast. Wines obtained were subjected to chemical and sensory analysis. The comparison between the two products highlighted differences due to the fermenting strains employed. The vineyard environment was found to strongly influence the composition of yeast communities, thus, confirming the theory of ‘terroir’ on the expression of wines. Moreover, vineyard inhabiting birds were in part responsible for the dissemination of fermentative yeasts during their feeding activities.     

Why are grape/fresh wine anthocyanins so simple and why is it that red wine color lasts so long?

Vitis vinifera red berries are characterized by anthocyanins whose chemical structures are among the simplest encountered in higher plants. On the contrary, many plants, including orchids, petunias, red cabbage, elderberries, potatoes for instance, have developed very complicated anthocyanins featuring side-chains at the available positions of the aglycone skeleton. Such pigments were shown to possess bio-physico-chemical properties not to be seen with the grape common anthocyanins. Among beverages (water, tea, beer, wine, coffee, juices, milk), red wine is the only one whose organoleptic properties improve with time and this is called ageing. The grape/fresh red wine pigments, after a few months, disappear from the wine giving birth to new pigments resulting from the wine spontaneous chemistry allowing it to remain red for many years. What are the wine pigments and why are they so stable is the purpose of this mini-review. The structural simplicity of grape anthocyanins and the long lasting colour of red wine is another French paradox; we call it French paradox II.     

Mössbauer and EPR study of iron in vacuoles from fermenting Saccharomyces cerevisiae

Vacuoles were isolated from fermenting yeast cells grown on minimal medium supplemented with 40 μM (57)Fe. Absolute concentrations of Fe, Cu, Zn, Mn, Ca, and P in isolated vacuoles were determined by ICP-MS. M?ssbauer spectra of isolated vacuoles were dominated by two spectral features: a mononuclear magnetically isolated high-spin (HS) Fe(III) species coordinated primarily by hard/ionic (mostly or exclusively oxygen) ligands and superparamagnetic Fe(III) oxyhydroxo nanoparticles. EPR spectra of isolated vacuoles exhibited a g(ave) ~ 4.3 signal typical of HS Fe(III) with E/D ~ 1/3. Chemical reduction of the HS Fe(III) species was possible, affording a M?ssbauer quadrupole doublet with parameters consistent with O/N ligation. Vacuolar spectral features were present in whole fermenting yeast cells; however, quantitative comparisons indicated that Fe leaches out of vacuoles during isolation. The in vivo vacuolar Fe concentration was estimated to be ~1.2 mM while the Fe concentration of isolated vacuoles was ~220 μM. M?ssbauer analysis of Fe(III) polyphosphate exhibited properties similar to those of vacuolar Fe. At the vacuolar pH of 5, Fe(III) polyphosphate was magnetically isolated, while at pH 7, it formed nanoparticles. This pH-dependent conversion was reversible. Fe(III) polyphosphate could also be reduced to the Fe(II) state, affording similar M?ssbauer parameters to that of reduced vacuolar Fe. These results are insufficient to identify the exact coordination environment of the Fe(III) species in vacuoles, but they suggest a complex closely related to Fe(III) polyphosphate. A model for Fe trafficking into/out of yeast vacuoles is proposed.     

Is vegetative area,photosynthesis, or grape C uploading involved in the climate change-related grape sugar/anthocyanin decoupling in Tempranillo?

Foreseen climate change is expected to impact on grape composition, both sugar and pigment content. We tested the hypothesis that interactions between main factors associated with climate change (elevated CO₂, elevated temperature, and water deficit) decouple sugars and anthocyanins, and explored the possible involvement of vegetative area, photosynthesis, and grape C uploading on the decoupling. Tempranillo grapevine fruit-bearing cuttings were exposed to CO₂ (700 vs. 400 ppm), temperature (ambient vs. +?4 °C), and irrigation levels (partial vs. full) in temperature-gradient greenhouses. In a search for mechanistic insights into the underlying processes, experiments 1 and 2 were designed to maximize photosynthesis and enlarge leaf area range among treatments, whereas plant growth was manipulated in order to deliberately down-regulate photosynthesis and control vegetative area in experiments 3 and 4. Towards this aim, treatments were applied either from fruit set to maturity with free vegetation and fully irrigated or at 5–8% of pot capacity (experiments 1 and 2), or from veraison to maturity with controlled vegetation and fully irrigated or at 40% of pot capacity (experiments 3 and 4). Modification of air ¹³C isotopic composition under elevated CO₂ enabled the further characterization of whole C fixation period and C partitioning to grapes. Increases of the grape sugars-to-anthocyanins ratio were highly and positively correlated with photosynthesis and grape ¹³C labeling, but not with vegetative area. Evidence is presented for photosynthesis, from fruit set to veraison, and grape C uploading, from veraison to maturity, as key processes involved in the establishment and development, respectively, of the grape sugars to anthocyanins decoupling.     

Badister Clairville 1806: A new species and new continental record for the nominate subgenus in Amazonian Perú (Coleoptera, Carabidae, Licinini)

Badister (Badister) amazonussp. n. is described from Perú, Loreto, 1.0 km SW Boca del Rio Samiria, Vigilante Post 1, 130m, "04°40.5'S, 074°18.9'W" its type locality. It is known also from two other localities in Loreto Department, Perú, in both the Varzea and Igapó river systems. This new species is sufficiently different that a new informal higher taxon, the amazonus species complex, is recognized. An updated key to the Western Hemisphere species of subgenus Badister is provided.     

A new species of Elasmia Möschler from New Mexico and Texas, and a new subspecies of Elasmia mandela (Druce) from Texas and Oklahoma (Lepidoptera, Notodontidae, Nystaleinae)

Hippia packardii (Morrison) and Hippia insularis (Grote) are moved to the genus Elasmia Möschler as comb. n. Elasmia cave Metzler, sp. n. is described from New Mexico and Texas, and Elasmia mandela santaana Metzler & Knudson, ssp. n. is described from Texas and Oklahoma. A key to the species of Elasmia of southwestern U.S. is provided. Adult male and female moths of Elasmia from southwestern U.S. and their genitalia are illustrated.     

Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice

Background   

Reporter genes are widely used in biology and only a limited number are available. We present a new reporter gene for the localization of mammalian cells and transgenic tissues based on detection of the bglA (SYNbglA) gene of Caldocellum saccharolyticum that encodes a thermophilic β-glucosidase.     

Interactions between Asplanchna and Keratella cochlearis in the Plußsee (north Germany)

Extremely high abundance of Asplanchna priodonta led to a decline in the population of the preferred food species, Keratella cochlearis and subsequently, to the extinction of the predator population. Kellicottia longispina was obviously favoured by the predatory losses of the Keratella. Thus, the interactions between Asplanchna and Keratella influenced the zooplankton community structure.     

9-(2-Deoxy-ß-D-xylofuranosyl)adenine and 1-(2-Deoxy-ß-D-xylofuranosyl)thymine: Phosphorylation and Stability

Abstract

Phosphorylation of 1-(2-deoxy-β-D-xylofuranosyl)thymine (1) or 9-(2-deoxy-β-D-xylofuranosyl)adenine (3) with phosphoryl chloride gives the cyclic 3′,5′-phosphates (2 and 4a) but not the 5′-monophosphates 8a or 8b. The latter are obtained by phosphorylation of the 3′-0-benzoylated 2′-deoxy-β-D-xylonucleosides (7a, b) and subsequent base-catalyzed removal of the benzoyl groups. Compound 3, as the parent dA, depurinates in acidic medium, a reaction which is facilitated in the case of the N⁶-benzoyl derivative 9b and reduced after the introduction of an amidine protecting group. N-Glycosylic bond hydrolysis of 2′-deoxy-β-D-xylofuranosyl nucleosides is enhanced by a factor of two compared to 2′-deoxy-β-D-ribofuranosyl nucleosides.     

Ocurrence of Cryptoblabes gnidiella (Millière) (Lepidoptera: Pyralidae) associated with grape phenology in Bento Gonçalves, RS

The aim of this study was to record and compare the occurrence of adults and immature stages of honeydew moth, Cryptoblabes gnidiella (Millière), related with grape phenology in two Pinot Noir (Vitis vinifera) vineyards in Bento Gon?alves (29 degrees 10'S 51 degrees 32'O), RS, during 2004-2005. Fortnightly, immature stages were monitored in the clusters and grapevine branches, while the adults, in delta traps baited with synthetic sex pheromone. There were significant differences in the number of individuals in different phenologic stages of vineyards. The highest catches of honeydew moth were registered in the period of dry clusters. The permanence of dry cluster after harvest, could serve as refuge for C. gnidiella immatures between seasons.