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PubMed2006

Cloning, functional identification and structural modelling of Vitis vinifera S-adenosylmethionine decarboxylase.

In this paper we report the cloning and full sequencing of S-adenosylmethionine decarboxylase (SAMDC, EC 4.1.1.50) cDNA from Vitis vinifera L. (VV) leaves, an enzyme belonging to the polyamine biosynthetic pathway, which appears to play an important role in the regulation of plant growth and development. The presence of two overlapping ORFs (tiny ORF and small ORF) upstream of the main ORF is reported in the Vitis cDNA. When the Vitis SAMDC cDNA was expressed in yeast without the two upstream ORFs, the resulting activity was about 50 times higher than the activity obtained with the full cDNA. These results demonstrated the strong regulatory activity of the tiny and small ORFs. RT-PCR expression analysis showed evidence of a similar mRNA level in all the tissues tested, with the exception of the petioles. The VV SAMDC was also modelled using its homologues from Solanum tuberosum and Homo sapiens as template. The present work confirmed, for the first time in a woody plant of worldwide economic interest such as grapevine, the presence of a regulatory mechanism of SAMDC, enzyme that has a well-established importance in the modulation of plant growth and development.

Adenosylmethionine Decarboxylase↗
PubMed2004

Exposure to elevated carbon dioxide concentration in the dark lowers the respiration quotient of Vitis cane wood.

Cane cuttings of the grapevine rootstock Vitis rupestris Scheele x V. riparia Michx. cv. 3309 Couderc were brought out of endodormancy by warming at 30 degrees C. Cane pieces (12 to 13 cm long) with nodes containing a primary bud were placed in a gas exchange system and monitored for net respiratory fluxes of CO2 and O2. Grapevine respiration rates expressed on a wood volume basis were 1.4 to 3.4 mmol CO2 or O2 m-3s-1, which is higher than stem respiration rates reported for many other woody taxa but similar to rates measured for ecodormant buds of other Vitis species. Passive water loss from canes was 0.7 to 1.2 mmol H2O m-3s-1. During a 7-day period, nonstructural carbohydrate concentrations in cane wood declined only slightly, whereas sucrose was nearly completely consumed. When ambient CO2 concentration ([CO2]) was raised from 300 to 750 micro molmol-1 and then 2000 micromol mol-1, net CO2 exchange rates declined by 5.9 +/- 0.6 and then 11.0 +/- 0.6%, whereas net O2 consumption rates remained about constant. The mean respiration quotient (net CO2/O2 flux) for canes with intact ecodormant buds was 0.99 +/- 0.03 when the [CO2] was 300 micromol mol-1, and decreased to 0.87 +/- 0.03 and 0.088 +/- 0.02 when the [CO2] was increased to 750 and 2000 micromol mol-1, respectively. The results support the hypothesis that, in Vitis canes, inhibition of respiratory CO2 efflux in response to high [CO2] is an indirect consequence of non-photosynthetic carboxylation reactions, and not a result of inhibition of respiratory metabolism.

Carbon Dioxide↗
PubMed2006

A novel Bacteroidetes symbiont is localized in Scaphoideus titanus, the insect vector of Flavescence dorée in Vitis vinifera.

Flavescence dorée (FD) is a grapevine disease that afflicts several wine production areas in Europe, from Portugal to Serbia. FD is caused by a bacterium, "Candidatus Phytoplasma vitis," which is spread throughout the vineyards by a leafhopper, Scaphoideus titanus (Cicadellidae). After collection of S. titanus specimens from FD-contaminated vineyards in three different areas in the Piedmont region of Italy, we performed a survey to characterize the bacterial microflora associated with this insect. Using length heterogeneity PCR with universal primers for bacteria we identified a major peak associated with almost all of the individuals examined (both males and females). Characterization by denaturing gradient gel electrophoresis confirmed the presence of a major band that, after sequencing, showed a 97 to 99% identity with Bacteroidetes symbionts of the "Candidatus Cardinium hertigii" group. In addition, electron microscopy of tissues of S. titanus fed for 3 months on phytoplasma-infected grapevine plants showed bacterial cells with the typical morphology of "Ca. Cardinium hertigii." This endosymbiont, tentatively designated ST1-C, was found in the cytoplasm of previtellogenic and vitellogenic ovarian cells, in the follicle cells, and in the fat body and salivary glands. In addition, cell morphologies resembling those of "Ca. Phytoplasma vitis" were detected in the midgut, and specific PCR assays indicated the presence of the phytoplasma in the gut, fat body and salivary glands. These results indicate that ST1-C and "Ca. Phytoplasma vitis" have a complex life cycle in the body of S. titanus and are colocalized in different organs and tissues.

Animals↗
PubMed2005

Analysis and bioactive evaluation of the compounds absorbed into blood after oral administration of the extracts of Vaccinium vitis-idaea in rat.

In order to screen the active constituents of Vaccinium vitis-idaea L., the compounds absorbed into the rat blood after oral administration of ethanol extract of the stems and leaves of V. vitis-idaea (EEV) have been analyzed. Two compounds were detected in the plasma and identified as arbutin and fraxin, which are originally existed in the crude drug. Furthermore, the pharmacological effects of the two compounds involving EEV for curing acute and chronic respiratory tract infection were tested. The results showed that both arbutin and fraxin have the anti-inflammatory, anti-coughing and phlegm-removing effects. Therefore, it was ascertained that the arbutin and fraxin would be the main active constituents of V. vitis-idaea L.

Administration, Oral↗
PubMed2006

[Significant improved anthocyanins biosynthesis in suspension cultures of Vitis vinifera by process intensification].

The low-production is a ubiquitous problem and has prevented the commercialization of secondary metabolite production in plant cell culture. In order to examine the effective approaches to improvement of secondary metabolite production in plant cell culture, the investigation of anthocyanins accumulation in suspension cultures of Vitis vinifera, as a model system, had been initiated in our laboratory. In this present research, various elicitors and the precursor of phenylalanine were used in combination to enhance the anthocyanins production in suspension cultures of Vitis vinifera. And an integrated process with the combination of elicitation, precursor feeding and light irradiation was reported for rational bioprocess design. Among the combination treatment of phenylalanine feeding and several elicitors (methyl-beta-cyclodextrin, dextran T-40, methyl jasmonate, extracts of Aspergillus niger and Fusarium orthoceras), the combination with methyl jasmonate gave the highest anthocyanins production in suspension cultures of Vitis vinifera. When compared to the controls, the anthocyanins content (CV/g, FCW) and production (CV/L) increased by 2.7-fold and 3.4-fold, respectively. The optimum time for the addition of phenylalanine and methyl jasmonate was 4 days after inoculation. Two cell lines with different anthocyanins-producing capacity responded differently to the optimum combination treatment of 30 micromol/L phenylalanine feeding, 218 micromol/L methyl jasmonate elicitation and 3000 to approximately 4000 1x light illumination. The high-and low-anthocyanins-producing cell lines of VV05 and VV06 produced the maximum of 2975 and 4090 CV/L of anthocyanins that were 2.5- and 5.2-fold of the controls, respectively.

Acetates↗
PubMed1989

Vaginal intratubal insemination (VITI) and vaginal GIFT, endosonographic technique: early experience.

A new technique for vaginal intratubal insemination (VITI) and transvaginal gamete intra-Fallopian transfer (TV-GIFT) using endosonography has been developed. A total of 47 infertile couples were treated using a K-JITS 1000-1100 catheter (William Cook, Australia; Jansen Anderson Intratubal Transfer Set) introduced by Jansen and Anderson in Sydney, Australia, and endosonographic control with a Kretz Combison 310 echograph and a 5/7.5 MHz vaginal sectorial transducer. In 40 couples, the VITI technique was used, and in the other seven couples, TV-GIFT was used. Fifteen pregnancies (37.5%) were obtained in 63 VITI cycles (23.8% per treatment cycle) and three with TV-GIFT (42% per treatment cycle). A review is also presented of the migration--sedimentation technique used for management of male factor in an artificial insemination programme.

Adult↗
PubMed1998

The presence and characterization of a virF gene on Agrobacterium vitis Ti plasmids.

Octopine and nopaline strains of Agrobacterium tumefaciens differ in their ability to induce tumors on Nicotiana glauca. The presence of a virF locus on the octopine Ti plasmid makes N. glauca a host plant for these strains, indicating that the VirF protein is a host-range determinant. Here we show the presence of a virF locus not only on the Agrobacterium vitis octopine/cucumopine plasmids pTiAg57 and pTiTm4, but also on the nopaline Ti plasmids pTiAT1, pTiAT66a, and pTiAT66b. On the octopine Ti plasmids from A. tumefaciens the virF gene is located between the virE locus and the left border of the T-region. In contrast, the virF gene on Ti plasmids of A. vitis is located at the very left end of the vir-region near the virA locus. The virF gene of pTiAg57 has been sequenced and codes for a protein of 202 amino acids with a molecular mass of 22,280 Da. Comparison showed that the virF gene from A. vitis strain Ag57 is almost identical to that from A. tumefaciens octopine strains. The transcription of the pTiAg57 virF is inducible by the plant phenolic compound acetosyringone through the presence of a vir-box consensus sequence in its promoter region. The VirF protein from pTiAg57 can complement octopine A. tumefaciens strains deleted for virF as shown by tumor formation on N. glauca.

Agrobacterium tumefaciens↗
PubMed1997

Differential expression of chitinases in Vitis vinifera L. responding to systemic acquired resistance activators or fungal challenge.

The concept of systemic acquired resistance (SAR) enables a novel approach to crop protection, and particular pathogenesis-related proteins, i.e. an acidic chitinase, have been classified as markers of the SAR response. Basic class I (VCHIT1b) and a class III (VCH3) chitinase cDNAs were cloned from cultured Vitis vinifera L. cv Pinot Noir cells and used to probe the induction response of grapevine cells to salicylic acid or yeast elicitor. Furthermore, the cells were treated with the commercial SAR activators 2,6-dichloroiso-nicotinic acid or benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester. Elicitor or salicylic acid induced both VCHIT1b and VCH3 transcript abundances, whereas 2,6-dichloroiso-nicotinic acid or benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester enhanced exclusively the expression of VCH3. To assess the systemic sensation of chitinase expression, single leaves of Vitis vinifera L. cv Pinot Noir or Vitis rupestris plants were inoculated with Plasmopara viticola spore suspensions, and the VCH3 and VCHIT1b mRNA amounts in the infected versus the adjacent, healthy leaf were monitored. Two VCH3 mRNA maxima were observed 2 and 6 d postinoculation in the infected, susceptible V. vinifera tissue, whereas in the healthy leaf the transcript increased from low levels d 2 postinoculation to prominent levels d 6 to 8 postinoculation. The level of VCH3 mRNA increased also over 4 d in the inoculated, resistant V. rupestris tissue. However, necrotic spots rapidly limited the infection, and the VCH3 transcript was undetectable in the upper-stage, healthy leaf. The expression of VCHIT1b remained negligible under either experimental condition. Overall, the results suggest that the selective expression of VCH3 might be a reliable indicator of the SAR response in V. vinifera L.

Amino Acid Sequence↗
PubMed1997

Characterization of the Agrobacterium vitis pehA gene and comparison of the encoded polygalacturonase with the homologous enzymes from Erwinia carotovora and Ralstonia solanacearum.

DNA sequencing of the Agrobacterium vitis pehA gene revealed a predicted protein with an M(r) of 58,000 and significant similarity to the polygalacturonases of two other plant pathogens, Erwinia carotovora and Ralstonia (= Pseudomonas or Burkholderia) solanacearum. Sequencing of the N terminus of the PehA protein demonstrated cleavage of a 34-amino-acid signal peptide from pre-PehA. Mature PehA accumulated primarily in the periplasm of A. vitis and pehA+ Escherichia coli cells during exponential growth. A. vitis PehA released dimers, trimers, and monomers from polygalacturonic acid and caused less electrolyte leakage from potato tuber tissue than did the E. carotovora and R. solanacearum polygalacturonases.

Amino Acid Sequence↗
PubMed2006

Phylogenetic analyses of Vitis (Vitaceae) based on complete chloroplast genome sequences: effects of taxon sampling and phylogenetic methods on resolving relationships among rosids.

BACKGROUND: The Vitaceae (grape) is an economically important family of angiosperms whose phylogenetic placement is currently unresolved. Recent phylogenetic analyses based on one to several genes have suggested several alternative placements of this family, including sister to Caryophyllales, asterids, Saxifragales, Dilleniaceae or to rest of rosids, though support for these different results has been weak. There has been a recent interest in using complete chloroplast genome sequences for resolving phylogenetic relationships among angiosperms. These studies have clarified relationships among several major lineages but they have also emphasized the importance of taxon sampling and the effects of different phylogenetic methods for obtaining accurate phylogenies. We sequenced the complete chloroplast genome of Vitis vinifera and used these data to assess relationships among 27 angiosperms, including nine taxa of rosids. RESULTS: The Vitis vinifera chloroplast genome is 160,928 bp in length, including a pair of inverted repeats of 26,358 bp that are separated by small and large single copy regions of 19,065 bp and 89,147 bp, respectively. The gene content and order of Vitis is identical to many other unrearranged angiosperm chloroplast genomes, including tobacco. Phylogenetic analyses using maximum parsimony and maximum likelihood were performed on DNA sequences of 61 protein-coding genes for two datasets with 28 or 29 taxa, including eight or nine taxa from four of the seven currently recognized major clades of rosids. Parsimony and likelihood phylogenies of both data sets provide strong support for the placement of Vitaceae as sister to the remaining rosids. However, the position of the Myrtales and support for the monophyly of the eurosid I clade differs between the two data sets and the two methods of analysis. In parsimony analyses, the inclusion of Gossypium is necessary to obtain trees that support the monophyly of the eurosid I clade. However, maximum likelihood analyses place Cucumis as sister to the Myrtales and therefore do not support the monophyly of the eurosid I clade. CONCLUSION: Phylogenies based on DNA sequences from complete chloroplast genome sequences provide strong support for the position of the Vitaceae as the earliest diverging lineage of rosids. Our phylogenetic analyses support recent assertions that inadequate taxon sampling and incorrect model specification for concatenated multi-gene data sets can mislead phylogenetic inferences when using whole chloroplast genomes for phylogeny reconstruction.

Base Sequence↗
PubMed1999

Molecular analysis of a tryptophan-2-monooxygenase gene (IaaM) of Agrobacterium vitis.

Tryptophan-2-monooxygenase genes occur in a number of bacteria and encode the conversion of tryptophan to the plant hormone precursor indole-3-acetamide. The role of these genes in the plant-bacteria interaction is often unclear. However, their function as a virulence determinant is established for Pseudomonas savastanoi and Agrobacterium tumefaciens. Some members of the Agrobacteria, such as Agrobacterium vitis have a limited host range. We have characterized the tryptophan-2-monooxygenase (iaaM) gene of A. vitis strain AG162 and show it is different from other A. vitis strains and related to iaaM of A. rhizogenes. The sequence of AG162 iaaM was deposited in the Genbank database under the accession number AF142716.

Amino Acid Sequence↗
PubMed1999

Indigenous yeasts associated with two Vitis vinifera grape varieties cultured in southern Spain.

The yeast microbiota present on the surface of grapes of two Vitis vinifera varieties, Pedro Ximénez and Tempranillo de Rioja, grown in the Montilla-Moriles region of southern Spain was identified. The changes between veraison and the physiological ripeness time during 3 years were monitored. Overall, the yeast microbiota isolated was of oxidative metabolism. Sporobolomyces roseus and Cryptococcus albidus species occurred at all physiological stages, in the two Vitis vinifera varieties, and the three seasons studied. On the other hand, Kloeckera apiculata was never detected and Saccharomyces cerevisiae was scarcely isolated, it was only present, testimonial, in Tempranillo de Rioja grapes during the 1992 vintage. The widest variety of yeast species was observed in the 1992 season, and in contrast, the lowest number of species in both varieties of Vitis was detected in the 1994 season.

Climate↗
PubMed2000

Root pressure and specific conductivity in temperate lianas: exotic Celastrus orbiculatus (Celastraceae) vs. native Vitis riparia (Vitaceae).

The exotic temperate liana (woody vine) Celastrus orbiculatus has become a weed in Michigan, occurring in many of the same habitats as the native liana Vitis riparia. However, C. orbiculatus frequently develops into extensive monospecific infestations, while V. riparia does not. Freezing-induced embolism may be responsible for limiting liana distribution. Root pressure has been observed in numerous tropical lianas and temperate species of Vitis and has been implicated as vital to the recovery of xylem function in wide vessels following winter freezes. For both of these co-occurring lianas we investigated root pressure and water conductance as possible explanatory factors for their differential spread. According to our hypothesis, C. orbiculatus should have produced greater or more frequent root pressures than V. riparia. However, the reverse proved true, indicating that root pressure is not a prerequisite for weedy proliferation of C. orbiculatus. Additionally, the seasonal patterns of specific conductivity of stem xylem indicate that each species responds differently to environmental constraints. Vitis riparia establishes conductivity early in the growing season, before the leaves emerge, using root pressure to reverse embolism, but loses conductivity with the first freeze in early autumn. Celastrus orbiculatus is slow to establish conductivity, depending on new wood production, but leafs out sooner than V. riparia and maintains green leaves after the first freeze. Vulnerability curves of xylem to cavitation caused by water stress for the two species indicate that they respond similarly to dehydration. These results indicate that root pressures are not responsible for the invasive success of C. orbiculatus and suggest that other factors must be key to its prolific invasion.

Journal Article↗
PubMed2003

A microsatellite marker based framework linkage map of Vitis vinifera L.

We have constructed a framework linkage map based on microsatellite markers for Vitis vinifera L., the European wine grape. The mapping population consisted of 153 progeny plants from a cross of Vitis vinifera cvs. Riesling x Cabernet Sauvignon. One hundred fifty-two microsatellite markers and one polymorphic EST marker have been mapped to 20 linkage groups (2 n=38). The map covers 1,728 cM with an average distance between markers of 11.0 cM. Estimates of genome size, expected genome coverage, and observed genome coverage were determined with 135-140 markers. Genome length estimates differed between paternal and maternal data sets. Observed approximate genome coverage was 65% versus an expected coverage of 90%. Meiotic recombination rates were not significantly different between maternal and paternal parents. This map has been adopted as a reference map for the International Grape Genome Program.

Genetic Linkage↗
PubMed2006

Identification and molecular mapping of PdR1, a primary resistance gene to Pierce's disease in Vitis.

A major quantitative trait locus (QTL) controlling resistance to Pierce's disease (PD) of grape, caused by the bacterium Xylella fastidiosa (Xf), was identified on a Vitis linkage map and denoted as 'Pierce's disease resistance 1' (PdR1). Placement of the locus was accomplished by evaluating a family of full-sib progeny from a cross of two PD-resistant interspecific hybrids with resistance inherited from Vitis arizonica. Resistance was measured under greenhouse conditions by direct quantification of Xf numbers in stem tissues as well as by evaluation of disease symptoms based on leaf scorch and a cane maturation index (CMI). A large QTL (LOD 17.2) accounting for 72% of the phenotypic variance in bacterial numbers was localized to linkage group 14 of the male parent F8909-17. The approximate 95% confidence interval around the QTL peak extended 5.7 cM when using composite interval mapping. The other disease evaluation methods (leaf scorch and CMI, respectively) placed the resistance QTL to the same region on linkage group 14, although at wider 95% confidence intervals (6.0 and 7.5 cM), lower peak LOD scores (11.9 and 7.7) and accounting for less phenotypic variance (59 and 42%). This is the first report of an Xf resistance QTL mapped in any crop species. The relevance of the markers located in the region spanning the QTL will be discussed, addressing their usefulness for the development of PD-resistant grape cultivars.

Chromosome Mapping↗
PubMed2006

Ultrastructure and germination of Vitis vinifera cv. Loureiro pollen.

The cultivar Loureiro of Vitis vinifera is one of the most economically important, recommended in almost the totality of the Região Demarcada dos Vinhos Verdes. In vineyards, the grape productivity of this cultivar is normal while in others it is extremely low. The aim of this work was to study the morphology and germination of Vitis vinifera cv. Loureiro pollen with high and low productivity. The pollen grain was examined under light, transmission and scanning electron microscopy. Typically V. vinifera pollen present three furrows but in the cultivar Loureiro we found tricolporated and acolporated (without furrows or pores) pollen grains. Both pollen types present generative and vegetative cells with the usual aspect and a dense cytoplasm rich in organelles. In the acolporated pollen a continuous exine layer and an irregular intine layer were observed. Differences were found in the starch accumulation, since only in tricolporated pollen abundant plastids filled with numerous starch granules were observed. To determine the causes of the low productivity of this cultivar we tested pollen viability by the fluorochromatic reaction and pollen germinability by in vitro assays. We observed that the acolporated pollen grain is viable, but no germination was recorded.

Cell Survival↗
PubMed2006

Evaluation of transgenic 'Chardonnay' (Vitis vinifera) containing magainin genes for resistance to crown gall and powdery mildew.

Magainins, short peptides with broad-spectrum antimicrobial activity in vitro, were assayed for their ability to confer resistance to pathogens in transgenic grapevines. Embryogenic cell suspensions of 'Chardonnay' (Vitis vinifera L.) were co-transformed by microprojectile bombardment with a plasmid carrying the npt-II gene and a second plasmid harboring either a natural magainin-2 (mag2) or a synthetic derivative (MSI99) gene. Magainin genes and the marker gene were driven by Arabidopsis ubiquitin-3 and ubiquitin-11 promoters, respectively. A total of 10 mag2 and 9 MSI99 regenerated lines were studied by Southern blot hybridization, which showed 1-6 transgene integration events into the plant genome. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed a variable range in transcription levels among mag2 and MSI99 lines. A positive correlation between number of integration events and transcription level was observed (p<0.05). Plants were acclimated and challenged in the greenhouse with either Agrobacterium vitis strains (bacterial crown gall pathogen) at 10(8) cfu/ml or Uncinula necator (fungal powdery mildew pathogen) at 10(5) conidia/ml for evaluation of disease resistance. A total of 6 mag2 and 5 MSI99 lines expressing the antimicrobial genes exhibited significant reductions of crown gall symptoms as compared to non-transformed controls. However, only two mag2 lines showed measurable symptom reductions in response to U. necator, but not strong resistance. Our results suggest that the expression of magainin-type genes in grapevines may be more effective against bacteria than fungi. Additional strategies to enhance transgene expression and the spectrum of resistance to grape diseases are suggested.

Amino Acid Sequence↗
PubMed2003

Adaptive responses to iron-deficiency in callus cultures of two cultivars of Vitis spp.

The correlation between iron chlorosis resistance and induction of adaptive mechanisms in grapevine calli belonging to cultivars with different susceptibility to iron chlorosis has been investigated. Fe(III)-chelate reductase was clearly linked to the Fe-efficiency status of the genotype. When growing on iron deprived medium (-Fe) calli of the Fe-efficient genotype "Cabernet sauvignon" showed a remarkable increase in enzyme activity, up to five times higher, with respect to +Fe cultures. Moreover, 31P-NMR revealed that in -Fe medium the increase of vacuolar Pi content of the Fe-efficient cultures was more pronounced than that recorded for the Fe-inefficient Vitis riparia. Furthermore, Fe starvation also enhanced the production of phenolic compounds in calli of "Cabernet sauvignon" with respect to those of Vitis riparia. The role of H(+)-ATPase as a marker of Fe-efficiency in tissue culture remains ambiguous in the case of grapevines.

Adaptation, Physiological↗