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Long-term experimental warming, shading and nutrient addition affect the concentration of phenolic compounds in arctic-alpine deciduous and evergreen dwarf shrubs.

Environmental changes are likely to alter the chemical composition of plant tissues, including content and concentrations of secondary compounds, and thereby affect the food sources of herbivores. After 10 years of experimental increase of temperature, nutrient levels and light attenuation in a sub-arctic, alpine ecosystem, we investigated the effects on carbon based secondary compounds (CBSC) and nitrogen in one dominant deciduous dwarf shrub, Salix herbacea x polaris and two dominant evergreen dwarf shrubs, Cassiope tetragona and Vaccinium vitis-idaea throughout one growing season. The main aims were to compare the seasonal course and treatment effects on CBSC among the species, life forms and leaf cohorts and to examine whether the responses in different CBSC were consistent across compounds. The changes in leaf chemistry both during the season and in response to the treatments were higher in S. herbacea x polaris than in the corresponding current year's leaf cohort of the evergreen C. tetragona. The changes were also much higher than in the 1-year-old leaves of the two evergreens probably due to differences in dilution and turnover of CBSC in growing and mature leaves paired with different rates of allocation. Most low molecular weight phenolics in the current year's leaves decreased in all treatments. Condensed tannins and the tannin-to-N ratio, however, either increased or decreased, and the strength and even direction of the responses varied among the species and leaf cohorts, supporting views of influential factors additional to resource-based or developmental controls, as e.g. species specific or genetic controls of CBSC. The results indicate that there is no common response to environmental changes across species and substances. However, the pronounced treatment responses imply that the quality of the herbivore forage is likely to be strongly affected in a changing arctic environment, although both the direction and strength of the responses will be different among plant species, tissue types and substances.

Arctic Regions↗

Analytical methods for monoterpene glycosides in grape and wine. II. Qualitative and quantitative determination of monoterpene glycosides in grape.

Free and glycosidically bound terpenes of five Vitis vinifera grape cultivars (muscat of Alexandria, muscat of Frontignan, muscat of Hamburg, muscat Ottonel and Gewürztraminer) were investigated. The free and bound fractions were separated by selective retention on Amberlite XAD-2 resin. The glycosidic fractions were analysed by gas chromatography and gas chromatography-mass spectrometry using either enzymic hydrolysis and subsequent analysis of the released aglycones or trimethylsilyl (TMS) and trifluoroacetyl derivatives. The known monoterpenyl, benzyl and 2-phenylethyl beta-D-glucopyranosides, beta-rutinosides, 6-O-alpha-L-arabinofuranosyl-beta-D-glucopyranosides and 6-O-beta-D-apiofuranosyl-beta-D-glucopyranosides were determined. A number of other glycosides were detected and the structures of some of them, mainly apiosylglucosides and glucosides with aglycones in higher oxidation state than linalol, were tentatively identified using the mass spectra of their TMS and TFA derivatives and the results obtained from the analysis of their aglycones.

Chromatography, Gas↗

Separation of viroid RNAs by cellulose chromatography indicating conformational distinctions.

A series of viroids and viroid-like RNAs extracted from citron (Citrus medica), grapevine (Vitis vinifera), and avocado (Persea americanum) displayed differential binding capacities to cellulose in the presence of ethanol. This nonionic interaction was influenced by the presence of magnesium ions suggesting variations in conformation among the viroid RNAs and corresponding differences in reactivity with cellulose. The specific elution profiles present a basis for the subclassification of groups of viroid-like molecules.

Cellulose↗

Volatilization of selenium from agricultural evaporation pond sediments.

Microbial volatilization of Se was evaluated as a means of detoxifying Se-contaminated sediments. Sediment samples containing 60.7 (Kesterson Reservoir) and 9.0 mg Se kg-1 (Peck ponds) were incubated for 273 days in closed systems located in the greenhouse. Volatile Se was collected from a continuous air-exchange stream using activated carbon. Various economical and readily available organic and inorganic amendments were tested for their capacity to enhance the microbial process, including Citrus (orange) peel, Vitis (grape) pomace, feedlot manure, barley straw, chitin, pectin, ZnSO4, (NH4)2SO4, and an inoculum of Acremonium falciforme (an active Se methylating fungus). With the Kesterson sediment, the highest Se removal (44.0%) resulted from the combined application of citrus peel and ZnSO4, followed by citrus peal alone (39.6%), and citrus peel combined with ZnSO4, (NH4)2SO4 and A. falciforme (30.1%). Manure (19.5%), pectin (16.4%), chitin (9.8%) and straw plus N (8.8%) had less pronounced effects. Without the amendments, cumulative Se volatilization was 6.1% of the initial inventory. Grape pomace (3.0%) inhibited the process. With the Peck sediment, the highest amount of Se removed was observed with chitin (28.6%), manure (28.5%), and citrus peel alone (27.3%). Without amendments, 14.0% of the native Se was volatilized in 273 days. Cumulative Se volatilization was 24.7% with citrus plus Zn and N, 17.2% with citrus plus Zn, and 18.8% with citrus plus Zn, N and A. falciforme. Pectin (15.2%), straw plus N (16.4%), and grape pomace (7.3%) were among the less effective amendments for the Peck sediment. The differences in the effectiveness of each treatment between the two seleniferous soils may be a result of the residual N content of the sediments. With the Kesterson sediment, which was high in organic C and N, added N inhibited volatilization of Se, while with Peck sediments (low in organic C and N) N-rich materials tended to accelerate Se volatilization. Inoculation with A. falciforme did not enhance Se evolution from either sediment, indicating that there was a sufficient population of microflora capable of producing gaseous Se.

Acremonium↗

Physical map of the vitopine Ti plasmid pTiS4.

Within the Agrobacterium vitis group the vitopine strains represent a special subclass. Vitopine bacteria carry Ti plasmids with little or no homology with the well-characterized T-DNAs of Agrobacterium tumefaciens or Agrobacterium rhizogenes. The 262-kb Ti plasmid of the vitopine strain S4 was cloned and mapped. Homology studies with the octopine Ti plasmid pTiAch5, the nopaline Ti plasmid pTiC58, and the agropine/mannopine Ri plasmid pRiHRI identified several regions of homology. The origin of replication was localized to within 2.5 kb.

Drug Resistance, Microbial↗

Mutagen content of table wines made from various grape species and hybrid cultivars.

The mutagen content of wines produced from the European Vitis vinifera grapes was compared with that of wines produced from hybrids of V. vinifera and species indigenous to North America. Mutagens were extracted on an XAD-2 Amberlite resin column and activated with S-9 and/or faecalase in the Salmonella/microsomal mutagen assay. All white wines had insignificant mutagen levels. The only red wine to produce statistically significant reversion frequencies was that made from the Concord grape. The mutagens were shown to be extracted from the grape skins during fermentation.

Fermentation↗

Oxidation of hydroquinone by both cellular and extracellular grapevine peroxidase fractions.

The oxidation of hydroquinone by two peroxidase (EC 1.11.1.7) fractions obtained from the cells and spent medium of cell cultures of grapevine (Vitis vinifera cv Monastrell) has been studied, and their comparative efficacy (kcat/KM ratio) studied in both the H2O2-consuming and hydroquinone-consuming reactions. While the efficacy in the H2O2-consuming reaction is practically identical for both enzyme fractions, the cellular peroxidase has five-fold more efficacy in the hydroquinone-consuming reaction than the peroxidase located in the spent medium. Screening of cellular peroxidases capable of oxidizing hydroquinone on polyacrylamide gels, by means of a staining reaction based on the nucleophilic attack of 4-aminoantipyrine on p-benzoquinone in acidic media, reveals that all the cellular peroxidase isoenzymes are capable of oxidizing hydroquinone, probably yielding a quinone-diimine as a product of the staining reaction. Since isoperoxidases found in cellular fractions are also present in the spent medium, the values found for the different efficacies in the hydroquinone-consuming reaction must be considered as the results of the different proportions in which each peroxidase isoenzyme was found in the two fractions. The localization of a benzoquinone-generating system of high efficacy inside the plant cell, and probably located in vacuoles, is discussed with respect to the harmful role which the quinone/semiquinone pair might play in cell death, as part of the hypersensitive response expressed within the mechanism of plant disease resistance.

Ampyrone↗

The amino-acid sequence similarity of plant glutamate dehydrogenase to the extremophilic archaeal enzyme conforms to its stress-related function.

A cDNA clone encoding grapevine (Vitis vinifera L. cv Sultanina) NAD(H)-glutamate dehydrogenase (GDH) was isolated from a cDNA expression library by immunoscreening with a polyclonal antibody raised against grapevine GDH. Nucleotide sequence analysis revealed an open reading frame (ORF) encoding a precursor protein of 411 amino acids (aa) with a calculated molecular mass of 44.517 kDa. The deduced aa sequence showed relatively higher homology to GDH from archaebacteria species, than to those from eukaryotes and eubacteria. This resemblance indicated a functional and/or evolutionary relationship in this class of enzymes which might be relevant to the stress-related function of plant GDH. We have shown that the bacterially produced plant GDH was thermostable.

Amino Acid Sequence↗

Evaluation of anti-inflammatory activity of some Swedish medicinal plants. Inhibition of prostaglandin biosynthesis and PAF-induced exocytosis.

Plants used in Swedish traditional medicine to treat inflammatory diseases and/or wounds were selected, based on literature data, for evaluation of inhibitory activity on prostaglandin biosynthesis and platelet activating factor (PAF)-induced exocytosis in vitro. Fifty-nine water extracts from 52 different plants in 28 families were tested. A number of plants, e.g. Calluna vulgaris, Corylus avellana, Geum urbanum, Juniperus communis, Polygonum aviculare, Potentilla erecta and Salix caprea were found to be active in both assays. The most potent cyclooxygenase inhibitors were extracts of Calluna vulgaris, Potentilla erecta and Salix caprea. None of the extracts inhibited just the prostaglandin biosynthesis. In the PAF-test, high inhibition was obtained by 19 extracts, the most potent of which were from Geum rivale, G. urbanum, Solanum dulcamara, Symphytum x uplandicum and Vaccinium vitis-idaea. The in vitro effects in relation to the traditional use, chemical contents and botanical classification, as well as the possibilities and the limitations of the methods are discussed.

Animals↗

Oxidative stress in recalcitrant tissue cultures of grapevine.

Thiobarbituric acid reactive substances (TBARS), and fluorescent compounds with spectral characteristics typical of products associated with oxidative stress in senescent and aging plant and animal cells, were detected in tissue cultures of the recalcitrant grapevine Vitis vinifera L. cultivar, Sultanina. These compounds increased during the early stages of dedifferentiation (callogenesis) of nodal stem explants. Catalase activity was not detected in the original explant, but was induced during callogenic dedifferentiation. Conversely, superoxide dismutase activity was detectable in the original explant, but diminished during the first week of callus induction. Transfer to callus induction medium promoted a large increase in the sulfhydryl content of nodal tissues. TBARS and fluorescent products accumulated in Sultanina callus during long-term culture (over 6 months). The possibility that oxidative stress may contribute to culture recalcitrance in this vine is discussed.

Agriculture↗

[Determination of an ELISA test on tissues of grapevine affected by yellow disease].

We report here results concerning the best part of the grapevine to section, the best sampling period and an important adaptation of the extraction media for the diagnosis of a grapevine yellow disease by ELISA. The addition of Triton-X100, or better of Chaps enabled us to get clear results for the sensitive Vitis vinifera scion varieties. Until now, results have not been obtained for the symptomless root stock varieties. However, antigen concentration by molecular filtration may be a way to solve the problem of assaying root stocks.

Detergents↗

Pectin methylesterases: sequence-structural features and phylogenetic relationships.

Pectin methylesterases (PMEs) are enzymes produced by bacteria, fungi and higher plants. They belong to the carbohydrate esterase family CE-8. This study deals with comparison of 127 amino acid sequences of this family containing the five characteristic sequence segments: 44_GxYxE, 113_QAVAL, 135_QDTL, 157_DFIFG, 223_LGRPW (Daucus carota numbering). Six strictly conserved residues (Gly44, Gly154, Asp157, Gly161, Arg225 and Trp227) and six conservative ones (Ile39, Ser86, Ser137, Ile152, Ile159 and Leu223) were identified. A set of 70 representative PMEs was created. The sequences were aligned and the evolutionary tree based on the alignment was calculated. The tree reflected the taxonomy: the fungal and bacterial PMEs formed their own clusters and the plant enzymes were grouped into eight separate clades. The plant PME from Vitis riparia was placed in a common clade with fungi. Three plant clades (Plant 1, 2 and 3) were relatively homogenous reflecting high degree of mutual sequence identity. The clade Plant 4 contained PMEs from flower parts (mostly form pollen) and was heterogenous, like the clades Plant 1a and 2a, which moreover exhibit an intermediate character. The clades Plant X1 and X2 were situated in the tree close to microbial clades and represented atypical plant PMEs. Taking into account the remaining plant PMEs, an expanded plant alignment and tree (with most Arabidopsis thaliana and Oryza sativa enzymes), were prepared. An exclusive Arabidopsis alignment and tree indicated the existence of a new plant clade X3. In the pre pro region of most plant enzymes a longer conserved segment containing basic dipeptide, R(K)/R(K), that precedes the N-terminal end of PME was revealed. This was not observed in the clade Plant X1 and majority of the clade Plant X2. This study brings further the description of occurrence of potential glycosylation sites in pre pro sequences and in mature enzymes as well as important amino acid residues, such as aspartates, cysteines, histidines and other aromatic residues (Tyr, Phe and Trp), with discussion of their possible function in the activity of PMEs.

Amino Acid Sequence↗

Rapid tool for distinction of wines based on the global volatile signature.

This work describes a novel methodology for the rapid distinction of wines by headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry, followed by principal component analysis of the data (HS-SPME-GC-MS-PCA). Headspace SPME is used to extract and concentrate the volatile and semi-volatile fractions. A DB-FFAP fused silica GC capillary column of 30 m at 220 degrees C was used acting as a transfer line of the components sorbed by the Carbowax-divinylbenzene coating fibre to the mass spectrometer, which acts as a sensor (30<m/z<300). In this methodology, which does not require any pre-treatment of the sample, the global volatile signature of the wine headspace (chromatographic profile and m/z pattern of fragmentation in each scan) is evaluated without complete chromatographic separation of its components. In order to retrieve from the data as much chemical information as possible and to extract m/z fragments (markers) for the characterisation and distinction of the wines varieties, a PCA was applied to the data resultant from the unresolved volatile fraction. In the present study, two different monovarietal white wines (Vitis vinifera L. var. Fernão-Pires and Arinto) were tested. Associated to the fast character of the proposed methodology and robustness taking into account the extraction time, it is also important to focus the higher sensibility and the lower effect of the sample moisture of the MS sensor response when compared to the conventional e-noses.

Alcohols↗

Application of sample disruption methods in the extraction of anthocyanins from solid or semi-solid vegetable samples.

Cassis, a commercial powder obtained from blackcurrant juice (Ribes nigrum L.), was used to optimize the anthocyanin extraction by matrix solid-phase dispersion (MSPD) and by sea sand disruption method (SSDM). The optimum eluent conditions were 5.0 mL of MeOH/H2O (1:1, v/v) at pH 2. The extracts were analyzed by HPLC with diode-array detection (DAD) and the extraction efficiency was evaluated by statistical comparison (analysis of variance, ANOVA-single factor) of the compounds peak areas. The extraction efficiency for the MSPD procedure was dependent on the chemical nature of the C18 material but the optimized procedure yielded results similar to those obtained by the inexpensive sea sand disruption method (SSDM). This procedure also compared favorably to a previously published liquid-liquid extraction method. The optimized sea sand method was then used in the anthocyanin extraction from red grapes (Vitis vinifera L.) and strawberries (Fragaria ananassa D.) yielding chromatographic profiles similar to those reported in the literature for these products.

Anthocyanins↗

Headspace-solid phase microextraction-gas chromatography as a tool to define an index that establishes the retention capacity of the wine polymeric fraction towards ethyl esters.

A headspace-solid phase microextraction followed by gas chromatographic analysis (HS-SPME-GC) was developed to be applied in the study of the interactions between the wine polymeric fraction and the ethyl esters: ethyl hexanoate, ethyl octanoate, and ethyl decanoate. Wine models (WM) were prepared with 10% (v/v) aqueous ethanol at pH 3.5 with distinct wine polymeric concentrations prepared from white wine of Vitis vinifera L. var. Fernão-Pires: 1.0 g L(-1) (PWM1), with a polymeric concentration approaching the real one in wine; 10.0 g L(-1) (PWM10); and 30.0 g L(-1) (PWM30), saturated with polymeric fraction. A reference wine model (RWM) was prepared without polymeric fraction. Each volatile compound (4.0 mg L(-1)) was added separately to the RWM and to the WM with the three levels of polymeric material (PWM). From the retention index (RI) calculated for each compound using the formula: [RI = 1 - (C(RWM) - C(PWM))/C(RWM)], where C(RWM) is the concentration of the compound in the RWM and C(PWM) is the concentration of the compound in the given PWM, the retention capacity of each wine polymeric fraction towards the three esters was established. The higher retention indexes were observed for ethyl decanoate, the more hydrophobic compound, and for the PWM with higher concentration. Furthermore, this study also suggested that the retained compounds are dosed to the headspace, which may promote the perception of their aroma for a longer period of time.

Adsorption↗

Eastern origin and three-millennia persistence of a founding grapevine lineage in Iberian viticulture.

Viticulture became central to most Mediterranean civilizations a few millennia after the grapevine (Vitis vinifera L.) was domesticated in the South Caucasus/Near East. To elucidate the origins of the grapevines that enabled this westward spread over the past 3,000 years, we analyzed 28 grapevine seeds from seven archaeological sites in the Iberian Peninsula and Sardinia. Ancient DNA recovered from the oldest seeds with domesticated-like morphology (from &#x223c;1,000 BCE), found in southwestern Spain, revealed nuclear and chloroplast genome signatures of Eastern Mediterranean cultivars. Seeds from the same and later Iron Age Iberian sites, however, showed genomic signatures suggesting hybridization between local wild grapevines and eastern-origin cultivars. The genetic makeup of Sardinian and northeastern Spanish seeds supports that local diversification giving rise to the Central European and Iberian wine genetic lineages had already occurred in the early Iron Age. In Iberia, Roman-period seeds were first-degree related to both the earliest eastern-introduced domesticates and a Medieval seed whose genetic makeup matches the extant Iberian variety "Pasa Valenciana." Another Medieval seed was inferred as an offspring of the extant "Heben," indicating that this major founder of Iberian germplasm has been continuously propagated for over 1,100 years. Our results confirm previous evolutionary models indicating that Western Mediterranean viticulture began with introductions of eastern domesticated grapevines, followed by early hybridization with local Iberian wild grapevines that may have facilitated viticulture adaptation to the new environment. The aDNA unveils that these introductions gave rise to extant cultivars through only a few sexual generations and long-term reliance on clonal propagation.

Iberian Peninsula↗

Responses to ammonium and nitrate additions by boreal plants and their natural enemies.

Separate effects of ammonium (NH4+) and nitrate (NO3-) on boreal forest understorey vegetation were investigated in an experiment where 12.5 and 50.0 kg nitrogen (N) ha(-1) year(-1) was added to 2 m2 sized plots during 4 years. The dwarf-shrubs dominating the plant community, Vaccinium myrtillus and V. vitis-idaea, took up little of the added N independent of the chemical form, and their growth did not respond to the N treatments. The grass Deschampsia flexuosa increased from the N additions and most so in response to NO3-. Bryophytes took up predominately NH4+ and there was a negative correlation between moss N concentration and abundance. Plant pathogenic fungi increased from the N additions, but showed no differences in response to the two N forms. Because the relative contribution of NH4+ and NO3- to the total N deposition on a regional scale can vary substantially, the N load a habitat can sustain without substantial changes in the biota should be set considering specific vegetation responses to the predominant N form in deposition.

Air Pollutants↗

Incidence of ozone symptoms on vegetation within a National Wildlife Refuge in New Jersey, USA.

During 1993-1996 and 2001-2003, we evaluated the percentage of plants (incidence) exhibiting ozone-induced foliar symptoms on vegetation within a National Wildlife Refuge located along the Atlantic Ocean coast of New Jersey, USA. Incidence varied among plant species and years. Bioindicator plants most sensitive to ozone, across all years, included native common milkweed (Asclepias syriaca) and wild grape (Vitis spp.), as well as introduced tree-of-heaven (Ailanthus altissima). Less sensitive bioindicators included Virginia creeper (Parthenocissus quinquefolia) and winged sumac (Rhus coppolina). Black cherry (Prunus serotina) and sassafras (Sassafras albidum) were least sensitive. The greatest incidence of ozone symptoms, across all plant species, occurred in 1996, followed by 2001>1995>1994>1993>2003>2002. A model was developed that showed a statistically significant relationship between incidence of ozone symptoms and the following parameters: plant species, Palmer Drought Severity Index, and the interaction of W126 x N100 measures of ambient ozone.

Air Pollutants↗