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Rapid decline in nitrate uptake and respiration with age in fine lateral roots of grape: implications for root efficiency and competitive effectiveness.

Changes in function as an individual root ages has important implications for understanding resource acquisition, competitive ability and optimal lifespan. Both nitrate uptake and respiration rates of differently aged fine roots of grape (Vitis rupestris x V. riparia cv. 3309 C) were measured. The resulting data were then used to simulate nitrate uptake efficiency and nutrient depletion as a function of root age. Both nitrate uptake and root respiration declined remarkably quickly with increasing root age. The decline in both N uptake and root respiration corresponded with a strong decline in root N concentration, suggesting translocation of nitrogen out of the roots. For simulations where no nutrient depletion occurs at the root surface, daily uptake efficiency was maximal at root birth and lifetime nitrate uptake efficiency slowly increased as the roots aged. Simulations of growth of roots into unoccupied soil using a solute transport model indicated the advantage of high uptake capacity in new roots under competitive conditions where nitrate availability is very transitory.

Nitrates↗

Canopy and environmental control of root dynamics in a long-term study of Concord grape.

Below-ground carbon allocation represents a substantial fraction of net photosynthesis in plants, yet timing of below-ground allocation and endogenous and exogenous factors controlling it are poorly understood. Minirhizotron techniques were used to examine root populations of Vitis labruscana Bailey cv. Concord under two levels of dormant-season canopy removal and irrigation. Root production, pigmentation, death and disappearance to a depth of 110 cm were determined over two wet and two dry years (1997-2000). There was continual root production and senescence, with peak root production rates occurring by midseason. Later in the season, when reproductive demands for carbon were highest and physical conditions limiting, few roots were produced, especially in dry years in nonirrigated vines. Root production under minimal canopy pruning was generally greater and occurred several weeks earlier than root production under heavy pruning, corresponding to earlier canopy development. Initial root production occurred in shallow soils, likely due to temperatures at shallow depths being warmer early in the season. Our study showed intricate relationships between internal carbon demands and environmental conditions regulating root allocation.

Agriculture↗

Characterization of a new, nonpathogenic mutant of Botrytis cinerea with impaired plant colonization capacity.

Botrytis cinerea is a necrotrophic pathogen that attacks more than 200 plant species. Here, the nonpathogenic mutant A336, obtained via insertional mutagenesis, was characterized. Mutant A336 was nonpathogenic on leaves and fruits, on intact and wounded tissue, while still able to penetrate the host plant. It grew normally in vitro on rich media but its conidiation pattern was altered. The mutant did not produce oxalic acid and exhibited a modified regulation of the production of some secreted proteins (acid protease 1 and endopolygalacturonase 1). Culture filtrates of the mutant triggered an important oxidative burst in grapevine (Vitis vinifera) suspension cells, and the mutant-plant interaction resulted in the formation of hypersensitive response-like necrosis. Genetic segregation analyses revealed that the pathogenicity phenotype was linked to a single locus, but showed that the mutated gene was not tagged by the plasmid pAN7-1. Mutant A336 is the first oxalate-deficient mutant to be described in B. cinerea and it differs from all the nonpathogenic B. cinerea mutants described to date.

Arabidopsis↗

Mercury hinders recovery of shoot hydraulic conductivity during grapevine rehydration: evidence from a whole-plant approach.

This experiment aimed to test whether recovery of shoot hydraulic conductivity after drought depends on cellular metabolism in addition to xylem hydraulics. We rehydrated droughted grapevines (Vitis vinifera) after treating intact plants through the root with 0.5 mm mercuric chloride (a metabolic inhibitor) at the end of the stress period, before rehydration. The contribution of mercury-inhibited water transport in both shoot and root, and the extent of shoot vessel embolization, were assessed. Drought stress decreased plant water potential and induced embolization of the shoot vessels. The rehydration in Hg-untreated plants re-established both shoot water potential and specific shoot hydraulic conductivity (Kss) at levels comparable with watered controls, and induced recovery of most of the embolisms formed in the shoot during the drought. In contrast, in plants treated with HgCl2, recovery of Kss and root hydraulic conductance were impaired. In rehydrated, Hg-treated plants, the effects of Hg on Kss were reversed when either the shoot or the root was treated with 60 mM beta-mercaptoethanol as a mercuric scavenger. This work suggests that plant cellular metabolism, sensitive to mercuric chloride, affects the recovery of shoot hydraulic conductivity during grapevine rehydration by interfering with embolism removal, and that it involves either the root or the shoot level.

Dehydration↗

Field distribution of Sorosphaera viticola in commercial vineyards in Germany.

In the year 2000, resting spores of a previously undescribed plasmodiophorid were found in roots of Vitis spp. This plasmodiophorid was identified as a member of the genus Sorosphaera Schroeter and described as Sorosphaera viticola Kirchmair, Neuhauser, Huber. To attain information on the field distribution of Sorosphaera viticola, a selective screening was conducted in two commercial vineyards in Germany. A study to determine a correlation of Sorosphaeraviticola infection to grapevine growth was also performed.

Myxomycetes↗

Enhancement of chilling resistance of inoculated grapevine plantlets with a plant growth-promoting rhizobacterium, Burkholderia phytofirmans strain PsJN.

In vitro inoculation of Vitis vinifera L. cv. Chardonnay explants with a plant growth-promoting rhizobacterium, Burkholderia phytofirmans strain PsJN, increased grapevine growth and physiological activity at a low temperature. There was a relationship between endophytic bacterial colonization of the grapevine plantlets and their growth at both ambient (26 degrees C) and low (4 degrees C) temperatures and their sensitivities to chilling. The major benefits of bacterization were observed on root growth (11.8- and 10.7-fold increases at 26 degrees C and 4 degrees C, respectively) and plantlet biomass (6- and 2.2-fold increases at 26 degrees C and 4 degrees C, respectively). The inoculation with PsJN also significantly improved plantlet cold tolerance compared to that of the nonbacterized control. In nonchilled plantlets, bacterization enhanced CO(2) fixation and O(2) evolution 1.3 and 2.2 times, respectively. The nonbacterized controls were more sensitive to exposure to low temperatures than were the bacterized plantlets, as indicated by several measured parameters. Moreover, relative to the noninoculated controls, bacterized plantlets had significantly increased levels of starch, proline, and phenolics. These increases correlated with the enhancement of cold tolerance of the grapevine plantlets. In summary, B. phytofirmans strain PsJN inoculation stimulates grapevine growth and improves its ability to withstand cold stress.

Acclimatization↗

Bleeding sap and old wood are the two main sources of contamination of merging organs of vine plants by Xylophilus ampelinus, the causal agent of bacterial necrosis.

The spatial distribution of vine plants contaminated by Xylophilus ampelinus, the agent responsible for bacterial necrosis, was studied over a 5-year period within two vineyards in the Cognac area. Both vineyards were planted with Vitis vinifera cv. Ugni blanc but were different in age and agronomic location. The emission of X. ampelinus in contaminated bleeding sap was observed during vine sprouting. Contaminated bleeding sap is an important source of inoculum for external contamination due to the high susceptibility of young merging shoots to the pathogen. X. ampelinus emission by bleeding sap was not affected by the age of the plants or the location of the vineyards. However, its emission was irregular with time, and it varied between two fruit canes from individual plants and between plants as well as between years. Moreover, the two vineyards appeared to be entirely contaminated. Consequently, the behavior of the pathogen is not predictable. The distribution of the pathogen inside vine plant organs was analyzed through the four growing seasons. The old wood was contaminated throughout the year and constituted a stock inoculum for endophytic contamination of crude sap during the winter and the spring. Despite the fact that most of the young green shoots were contaminated in May, X.ampelinus was not found in green shoots in June and September, refuting the hypothesis of an epiphytic life of the pathogen under natural conditions. Although all plants were entirely contaminated in both vineyards, symptoms were rare and were observed on different plants each year.

Burkholderiaceae↗

Asymptomatic sensitisation to grapes in a sample of workers in the wine industry.

AIMS: To assess the prevalence of sensitisation to grapes (Vitis vinifera var. agiorghitiko) in a population with repeated exposure to grape allergens through direct cutaneous contact as well as through the gastrointestinal tract. METHODS: One hundred and twenty subjects were enrolled in each of four groups: grape harvesters, winery workers in selection of grapes, winery workers operating de-stemming/crushing/pressing machines, and administrative personnel. Sensitisation to grapes was examined by skin prick-to-prick tests with fresh fruit and juice. RESULTS: Eight harvesters and five workers in grape selection had positive reaction to the grapes tested. No machine operators or administrative personnel had positive tests. The likelihood of sensitisation was estimated at 3.7% per year of occupation by logistic regression analysis. None of the employees reported symptoms associated with sensitisation to grapes. CONCLUSION: Asymptomatic sensitisation to grapes was detected only in workers handling the fruit, suggesting that sensitisation is more likely to occur through cutaneous exposure and/or minor wounding than through the gastrointestinal tract. Prevalence rates were high and the clinical impact needs to be further investigated.

Adult↗

An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development.

BACKGROUND: Accuracy in quantitative real-time RT-PCR is dependent on high quality RNA, consistent cDNA synthesis, and validated stable reference genes for data normalization. Reference genes used for normalization impact the results generated from expression studies and, hence, should be evaluated prior to use across samples and treatments. Few statistically validated reference genes have been reported in grapevine. Moreover, success in isolating high quality RNA from grapevine tissues is typically limiting due to low pH, and high polyphenolic and polysaccharide contents. RESULTS: We describe optimization of an RNA isolation procedure that compensates for the low pH found in grape berries and improves the ability of the RNA to precipitate. This procedure was tested on pericarp and seed developmental series, as well as steady-state leaf, root, and flower tissues. Additionally, the expression stability of actin, AP47 (clathrin-associated protein), cyclophilin, EF1-alpha (elongation factor 1-alpha), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), MDH (malate dehydrogenase), PP2A (protein phosphatase), SAND, TIP41, alpha-tubulin, beta-tubulin, UBC (ubiquitin conjugating enzyme), UBQ-L40 (ubiquitin L40) and UBQ10 (polyubiquitin) were evaluated on Vitis vinifera cv. Cabernet Sauvignon pericarp using three different statistical approaches. Although several of the genes proved to be relatively stable, no single gene outperformed all other genes in each of the three evaluation methods tested. Furthermore, the effect of using one reference gene versus normalizing to the geometric mean of several genes is presented for the expression of an aquaporin and a sucrose transporter over a developmental series. CONCLUSION: In order to quantify relative transcript abundances accurately using real-time RT-PCR, we recommend that combinations of several genes be used for normalization in grape berry development studies. Our data support GAPDH, actin, EF1-alpha and SAND as the most relevant reference genes for this purpose.

Aquaporins↗

Anthocyanin-rich extracts inhibit multiple biomarkers of colon cancer in rats.

The aim of the present study was to investigate the chemoprotective activity of anthocyanin-rich extracts (AREs) from bilberry (Vaccinium myrtillus L.), chokeberry (Aronia meloncarpa E.), and grape (Vitis vinifera) by assessing multiple biomarkers of colon cancer in male rats treated with a colon carcinogen, azoxymethane. Fischer 344 male rats were fed the AIN-93 diet (control) or AIN-93 diet supplemented with AREs for 14 wk. Biomarkers that were evaluated included the number and multiplicity of colonic aberrant crypt foci (ACF), colonic cell proliferation, urinary levels of oxidative DNA damage, and expression of cyclooxygenase (COX) genes. To assess the bioavailability, levels of anthocyanins in serum, urine, and feces were evaluated. Total ACF were reduced (P<0.05) in bilberry, chokeberry, and grape diet groups compared with the control group. The number of large ACF was also reduced (P<0.05) in bilberry and chokeberry ARE-fed rats. Colonic cellular proliferation was decreased in rats fed bilberry ARE and chokeberry ARE diets. Rats fed bilberry and grape ARE diets had lower COX-2 mRNA expression of gene. High levels of fecal anthocyanins and increased fecal mass and fecal moisture occurred in ARE-fed rats. There was also a significant reduction (P<0.05) in fecal bile acids in ARE-fed rats. The levels of urinary 8-hydroxyguanosine were similar among rats fed different diets. These results support our previous in vitro studies suggesting a protective role of AREs in colon carcinogenesis and indicate multiple mechanisms of action.

8-Hydroxy-2'-Deoxyguanosine↗

Antihypertensive, vasodilator and antioxidant effects of a vinifera grape skin extract.

Cumulative evidence suggests that moderate wine consumption exerts a cardioprotective effect. We investigated the occurrence of an antihypertensive effect of an alcohol-free hydroalcoholic grape skin extract (GSE) obtained from skins of a vinifera grape (Vitis labrusca) in experimental rodent hypertension models. The vasodilator effect of GSE (polyphenols concentration 55.5 mg g(-1)) was also assessed in the isolated mesenteric vascular bed of Wistar rats and the antioxidant effect was studied on lipid peroxidation of hepatic microsomes. Oral administration of GSE significantly reduced systolic, mean and diastolic arterial pressure in Wistar rats with desoxycorticosterone acetate-salt and N(G)-nitro-L-arginine methyl ester (L-NAME) induced experimental hypertension. In the rat isolated mesenteric vascular bed pre-contracted with norepinephrine, bolus injections of GSE induced endothelium-dependent vasodilatation that was substantially inhibited by L-NAME, but not by indometacin, tetraethylammonium or glibenclamide. Lipid peroxidation of hepatic microsomes estimated as malondialdehyde production was concentration-dependently inhibited by GSE. In conclusion, the antihypertensive effect of GSE might be owing to a combination of vasodilator and antioxidant actions of GSE. These findings also suggest that the beneficial effect of moderate red wine consumption could be owing to an antihypertensive action induced by compounds occurring in the skin of vinifera grapes.

Animals↗

Organ-specific transcription of putative flavonol synthase genes of grapevine and effects of plant hormones and shading on flavonol biosynthesis in grape berry skins.

In order to investigate the control mechanism of flavonol biosynthesis of grapevine, we obtained five genomic sequences (FLS1 to FLS5) of putative flavonol synthase genes from Vitis vinifera cv. Cabernet Sauvignon. The mRNA of five FLSs accumulated in flower buds and flowers, while the mRNA of FLS2, FLS4, and FLS5 accumulated in small berry skins and then decreased toward veraison. At the ripening stage, the mRNA of only FLS4 and FLS5 accumulated again. This change in mRNA accumulation did not contradict the flavonol accumulation in the berry skins. Shading of the berries completely inhibited the increase in flavonol content and mRNA accumulation of FLS4, but did not affect the mRNA accumulation of FLS5. The effects of light and plant hormones on flavonol accumulation were different from those on anthocyanin accumulation. Thus flavonol biosynthesis appears to be under a different control system from that of anthocyanin biosynthesis.

Amino Acid Sequence↗

Orthologs in Arabidopsis thaliana of the Hsp70 interacting protein Hip.

The Hsp70-interacting protein Hip binds to the adenosine triphosphatase domain of Hsp70, stabilizing it in the adenosine 5'-diphosphate-ligated conformation and promoting binding of target polypeptides. In mammalian cells, Hip is a component of the cytoplasmic chaperone heterocomplex that regulates signal transduction via interaction with hormone receptors and protein kinases. Analysis of the complete genome sequence of the model flowering plant Arabidopsis thaliana revealed 2 genes encoding Hip orthologs. The deduced sequence of AtHip-1 consists of 441 amino acid residues and is 42% identical to human Hip. AtHip-1 contains the same functional domains characterized in mammalian Hip, including an N-terminal dimerization domain, an acidic domain, 3 tetratricopeptide repeats flanked by a highly charged region, a series of degenerate GGMP repeats, and a C-terminal region similar to the Sti1/Hop/p60 protein. The deduced amino acid sequence of AtHip-2 consists of 380 amino acid residues. AtHip-2 consists of a truncated Hip-like domain that is 46% identical to human Hip, followed by a C-terminal domain related to thioredoxin. AtHip-2 is 63% identical to another Hip-thioredoxin protein recently identified in Vitis labrusca (grape). The truncated Hip domain in AtHip-2 includes the amino terminus, the acidic domain, and tetratricopeptide repeats with flanking charged region. Analyses of expressed sequence tag databases indicate that both AtHip-1 and AtHip-2 are expressed in A thaliana and that orthologs of Hip are also expressed widely in other plants. The similarity between AtHip-1 and its mammalian orthologs is consistent with a similar role in plant cells. The sequence of AtHip-2 suggests the possibility of additional unique chaperone functions.

Amino Acid Sequence↗

Cryopreservation of embryogenic cell suspensions by encapsulation-vitrification.

Encapsulation-vitrification, which is a combination of encapsulation-dehydration and vitrification procedures, is a newly developed technique for cryopreservation of plant germoplasm. Here, we describe the protocol of this methodology, using grapevine (Vitis) as a model plant. Cell suspensions at the exponential growth stage were encapsulated with 2.5% sodium alginate solution in 0.1 M calcium chloride solution for 20 min to form beads of about 4 mm in diameter containing 25% cells. The beads were stepwise precultured in increasing sucrose concentrations of 0.25, 0.5, and 0.75 M for 3 yr, with 1 d for each step. Precultured beads were treated with a loading solution for 60 min at room temperature and then dehydrated with PVS2 at 0 degrees C for 270 min, followed by direct immersion in liquid nitrogen for 1 h. The beads were rapidly rewarmed at 40 degrees C in a water bath for 3 min and then diluted with 1 M sucorse solution at room temperature for 30 min. Rewarmed, washed beads were post-cultured on a recovery medium for 3 d at 25 degrees C in the dark for survival. Surviving cells were transferred to a regrowth medium to induce cell proliferation. Embryogenic cell suspensions were re-established by suspending the cells in a cell suspension maintenance medium maintained on a gyratory shaker at 25 degrees C in the dark. For plant regeneration, surviving cells were transferred from the recovery medium to an embryo maturation medium and maintained at 25 degrees C under light conditions. Embryos at the torpedo stage were cultured on rooting medium until whole plantlet was developed.

Cell Survival↗

Mass spectrometry in the analysis of grape and wine proteins.

Wine proteins play an important role in the quality of wine, because they affect taste, clarity and stability of product. The majority of wine proteins are in the range of 20-30 kDa. Different mass spectrometry (MS) techniques have been successfully applied to study the grape and wine proteins. By liquid chromatography (LC) electrospray ionization (ESI) MS and nano-LC/MS, nine dipeptides and 80 peptides were unambiguously identified in Champagne and Sauvignon Blanc wines, respectively. Using matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) and surface-enhanced laser desorption/ionization TOF, the protein and peptide fingerprints in Chardonnay, Sauvignon Blanc and Muscat of Alexandria wines were determined. MALDI-TOF identified the mesocarp proteome of six Vitis grape varieties. Proteins in different grape tissue extracts were also studied. The major grape pathogenic-related proteins are chitinases and thaumatin-like proteins, which both persist through the vinification process and cause hazes and sediments in bottled wines. ESI-MS, LC/ESI-MS and MALDI-TOF analysis of these proteins in grape and wine were also used to characterize different grape varieties.

Food Industry↗

Application of competitive enzyme-linked immunosorbent assay for the quantification of imidacloprid titers in xylem fluid extracted from grapevines.

A competitive enzyme-linked immunosorbent assay (ELISA) technique was evaluated for quantifying titers of imidacloprid in xylem fluid extracted from Vitis vinifera L. grapevines that were treated with systemic applications of the neonicotinoid insecticide Admire. Evidence of matrix effects, factors that compromise the precision and accuracy of the ELISA, was present in assays with undiluted xylem fluid. These effects could be eliminated by dilution of extracts in water, resulting in a lower sensitivity of the assay of 4 microg liter(-1). In a field trial conducted in a commercial vineyard, there was an excellent correlation between Admire application rates and xylem fluid concentrations of imidacloprid. At an Admire application rate of 1.17 liter ha(-1) (16 fl oz per acre), uptake of imidacloprid into vines was rapid. Imidacloprid was consistently detected in the xylem for up to 3 mo after application at concentrations known to be effective at managing populations of the sharpshooter Homalodisca coagulata Say, an important vector of Xylella fastidiosa Wells in California vineyards. The ELISA is a sensitive technique that can be used to study the behavior of systemic insecticides within crop systems and their impact on pest populations.

Enzyme-Linked Immunosorbent Assay↗

Spatial distribution of nymphs of Scaphoideus titanus (Homoptera: Cicadellidae) in grapes, and evaluation of sequential sampling plans.

The spatial distribution of the nymphs of Scaphoideus titanus Ball (Homoptera Cicadellidae), the vector of grapevine flavescence dorée (Candidatus Phytoplasma vitis, 16Sr-V), was studied by applying Taylor's power law. Studies were conducted from 2002 to 2005, in organic and conventional vineyards of Piedmont, northern Italy. Minimum sample size and fixed precision level stop lines were calculated to develop appropriate sampling plans. Model validation was performed, using independent field data, by means of Resampling Validation of Sample Plans (RVSP) resampling software. The nymphal distribution, analyzed via Taylor's power law, was aggregated, with b = 1.49. A sample of 32 plants was adequate at low pest densities with a precision level of D0 = 0.30; but for a more accurate estimate (D0 = 0.10), the required sample size needs to be 292 plants. Green's fixed precision level stop lines seem to be more suitable for field sampling: RVSP simulations of this sampling plan showed precision levels very close to the desired levels. However, at a prefixed precision level of 0.10, sampling would become too time-consuming, whereas a precision level of 0.25 is easily achievable. How these results could influence the correct application of the compulsory control of S. titanus and Flavescence dorée in Italy is discussed.

Animals↗

Attenuation of renal ischemia-reperfusion injury by proanthocyanidin-rich extract from grape seeds.

The effects of proanthocyanidin-rich extract in rats subjected to renal ischemia-reperfusion were examined. Proanthocyanidin-rich extract, which is prepared from grape seeds (Vitis vinifera L.), was given orally at doses of 5 and 10 mg/kg body weight/d for 20 consecutive days prior to ischemia-reperfusion. Administration of proanthocyanidin-rich extract attenuated renal dysfunction, as indicated by serum urea nitrogen and creatinine levels. Additionally, in the ischemic-reperfused kidneys, increased levels of thiobarbituric acid (TBA)-reactive substance and alterations of antioxidant enzyme activities such as superoxide dismutase, catalase and glutathione peroxidase (GSH-Px) were observed. Proanthocyanidin-rich extract-treated groups showed significantly reduced renal TBA-reactive substance levels and enhanced catalase and GSH-Px activities. These results suggest that proanthocyanidin-rich extract has protective effects against ischemia-reperfusion-induced renal damage associated with oxidative stress.

Animals↗