Vitis Berlandieri Research Articles

Modulation of Protein Phosphorylation, N-Glycosylation and Lys-Acetylation in Grape (Vitis vinifera) Mesocarp and Exocarp Owing to Lobesia botrana Infection

Grapevine (Vitis vinifera) is an economically important fruit crop that is subject to many types of insect and pathogen attack. To better elucidate the plant response to Lobesia botrana pathogen infection, we initiated a global comparative proteomic study monitoring steady-state protein expression as well as changes in N-glycosylation, phosphorylation, and Lys-acetylation in control and infected mesocarp and exocarp from V. vinifera cv Italia. A multi-parallel, large-scale proteomic approach employing iTRAQ labeling prior to three peptide enrichment techniques followed by tandem mass spectrometry led to the identification of a total of 3059 proteins, 1135 phosphorylation sites, 323 N-linked glycosylation sites and 138 Lys-acetylation sites. Of these, we could identify changes in abundance of 899 proteins. The occupancy of 110 phosphorylation sites, 10 N-glycosylation sites and 20 Lys-acetylation sites differentially changed during L. botrana infection. Sequence consensus analysis for phosphorylation sites showed eight significant motifs, two of which containing up-regulated phosphopeptides (X-G-S-X and S-X-X-D) and two containing down-regulated phosphopeptides (R-X-X-S and S-D-X-E) in response to pathogen infection. Topographical distribution of phosphorylation sites within primary sequences reveal preferential phosphorylation at both the N- and C termini, and a clear preference for C-terminal phosphorylation in response to pathogen infection suggesting induction of region-specific kinase(s). Lys-acetylation analysis confirmed the consensus X-K-Y-X motif previously detected in mammals and revealed the importance of this modification in plant defense. The importance of N-linked protein glycosylation in plant response to biotic stimulus was evident by an up-regulated glycopeptide belonging to the disease resistance response protein 206. This study represents a substantial step toward the understanding of protein and PTMs-mediated plant-pathogen interaction shedding light on the mechanisms underlying the grape infection.

Phylogenetic Analyses of Teleki Grapevine Rootstocks Using Three Chloroplast DNA Markers

Teleki rootstocks are used in grapevine-producing countries all over the world. They represent one of the largest groups of available rootstocks but their origin is still in dispute although they have been regarded as Vitis berlandieri × V. riparia hybrids. To investigate their possible origin, we amplified and sequenced three chloroplast regions, two non-coding spacers (trnL-F, trnS-G) and the trnL group I intron in a core collection of Teleki rootstocks representing widespread accessions and related wild North American grape species (V. berlandieri, V. riparia and V. rupestris). Concatenated sequence data coupled with microstructural changes discovered in the chloroplast regions provided data to trace the maternal ancestry of the Teleki lines. All chloroplast regions showed both nucleotide and length variation. Length mutations in the non-coding regions represented mostly simple sequence repeats of poly-A and -T stretches. These indel characters exhibited additional diversity comparable with the nucleotide diversity and increased resolution of the phylogenetic trees. We found that a group of Teleki accessions position together with the wild grape species V. riparia. Another group of Teleki rootstocks formed a sister group to the other North American species V. berlandieri. These clades had moderate support values, and they do not share ancestry with other accessions of Teleki rootstocks resolved with high support value in the V. riparia clade. It seems that Teleki-Kober 5BB and 125 AA accessions might have a V. berlandieri maternal background. We also found great differences within putative clones of Teleki 5C and Teleki-Kober 5BB suggesting that the selection of these accessions was performed on heterogenous or mislabeled plant material collectively maintained under these names.

Agrobacterium tumefaciens virE1Inhibits crown gall development in transgenic grapevine

The biotic and abiotic stresses are the major limiting factors in plant productivity. To overcome these difficulties molecular breeding methods have recently been widely used to improve the stress and disease resistance of grapevine cultivars. Crown gall disease caused by Agrobacterium vitis or Agrobacterium tumefaciens causes serious damage worldwide on grapevine, and there is no efficient method yet that can be routinely used by grape-growers to prevent this disease. Therefore genetic manipulation for crown gall resistance would have a great economic impact. To this end embryogenic culture of Vitis berlandieri × Vitis rupestris cv. Richter 110 was transformed with a virE1 gene construct. Twenty-six plant lines were selected, and their transgenic nature was confirmed by PCR analysis. Seventeen of the 26 lines showed resistance to crown gall disease following inoculation with A. vitis Tm4 strain.

Screening of rootstock hybrids with Vitis cinerea Arnold for phylloxera resistance

AbstractThe resistance to phylloxera is one of the most important properties of grapevine rootstock. The breeding and selection of rootstock varieties represents a biological method of fight against phylloxera. In this study altogether 59 rootstock hybrids originating from nine different populations were evaluated. Pedigrees of all populations involved Vitis cinerea Arnold introduced through the hybrid Bruci/(Vitis berlandieri Planch. × Vitis rupestris Scheele) × Vitis cinerea Arnold/ and the variety Börner. Pot experiments were performed in a glasshouse while those with excised root pieces in root bioassay took place in a laboratory. This combination showed to be a very good selection tool for rootstock breeding. In seven rootstock hybrids, it was found that the resistance to phylloxera gallicoles and radicicoles was very high. The highest number of resistant hybrids was found in Binova × Börner. The variety Börner showed to be the best gene source of phylloxera resistance and even in case of a simple crossing produced sufficient number of resistant progeny.

Local and systemic mycorrhiza-induced protection against the ectoparasitic nematode Xiphinema index involves priming of defence gene responses in grapevine

The ectoparasitic dagger nematode (Xiphinema index), vector of Grapevine fanleaf virus (GFLV), provokes gall formation and can cause severe damage to the root system of grapevines. Mycorrhiza formation by Glomus (syn. Rhizophagus) intraradices BEG141 reduced both gall formation on roots of the grapevine rootstock SO4 (Vitis berlandieri×V. riparia) and nematode number in the surrounding soil. Suppressive effects increased with time and were greater when the nematode was post-inoculated rather than co-inoculated with the arbuscular mycorrhizal (AM) fungus. Using a split-root system, decreased X. index development was shown in mycorrhizal and non-mycorrhizal parts of mycorrhizal root systems, indicating that both local and systemic induced bioprotection mechanisms were active against the ectoparasitic nematode. Expression analyses of ESTs (expressed sequence tags) generated in an SSH (subtractive suppressive hybridization) library, representing plant genes up-regulated during mycorrhiza-induced control of X. index, and of described grapevine defence genes showed activation of chitinase 1b, pathogenesis-related 10, glutathione S-transferase, stilbene synthase 1, 5-enolpyruvyl shikimate-3-phosphate synthase, and a heat shock proein 70-interacting protein in association with the observed local and/or systemic induced bioprotection against the nematode. Overall, the data suggest priming of grapevine defence responses by the AM fungus and transmission of a plant-mediated signal to non-mycorrhizal tissues. Grapevine gene responses during AM-induced local and systemic bioprotection against X. index point to biological processes that are related either to direct effects on the nematode or to protection against nematode-imposed stress to maintain root tissue integrity.

Anatomy and osmotic potential of the Vitis rootstock shoot tips recalcitrant to cryopreservation

This study was carried out on Kober 5BB (Vitis Berlandieri × V. riparia) grape rootstock shoot tips during the preparatory steps preceding the direct immersion in liquid nitrogen, in order to overcome until now unsuccessful cryopreservation with this species. The exposure of shoot tips to 0.3-0.4 M sucrose leads to a high cell solute concentration. The treatment with plant vitrification solution (PVS2) alone, i.e., not followed by storage in liquid nitrogen, markedly affected shoot tip survival. After a 30 min exposure, regrowth percentage of shoot tips decreased from 94 % (control) to 57 %, and dropped to 15 % when the treatment was prolonged up to 60 min. After a 90 min exposure, no regrowth occurred. In addition, plantlets regenerated from shoot tips which underwent 60 min or more exposure to PVS2 showed signs of malformation. Microscope observations of shoot tips treated with 0.3 or 0.4 M sucrose and 30 min PVS2 showed the presence of cells starting to plasmolyze, localized in the area surrounding the apical meristem. A limited presence of starch grains in meristem and bract cells was also noted. However, the most conspicuous consequence of prolonged PVS2 treatment was convex plasmolysis. The phenomenon was dependent on the time of PVS2 exposure. Indeed, after a 30 min treatment, plasmolysis was minimal or absent, but it increased with longer exposure to PVS2 at 4 °C.

Grapevine rootstock effects on scion biomass are not associated with large modifications of primary shoot growth under nonlimiting conditions in the first year of growth.

In grapevine (Vitis vinifera L.), rootstocks are known to alter scion development by modifying stem weight and yield. The aim of this work was to evaluate the contribution of primary growth to the rootstock effects on scion biomass. The shoot growth of Vitis vinifera cv. Cabernet Sauvignon N autografted and grafted onto Vitis riparia cv. Riparia Gloire de Montpellier and Vitis berlandieri×V. rupestris cv. 1103 Paulsen was studied in young plants grown in pots trained to one stem in two experiments. Stem elongation and phytomer emergence were studied from grafting until the end of the growth season. The elongation of the Cabernet Sauvignon N leaves, tendrils and internodes of each phytomer along the stem was fitted using sigmoid curves. The rootstocks studied slightly altered the growth dynamics of the leaves, internodes and tendrils of the scion. This is the first study to examine the effect of rootstocks on shoot growth dynamics in any species. The alterations in primary growth were small, suggesting that rootstocks may alter scion biomass principally by modifying secondary growth.

Anatomy and osmotic potential of the Vitis rootstock shoot tips recalcitrant to cryopreservation

This study was carried out on Kober 5BB (Vitis Berlandieri × V. riparia) grape rootstock shoot tips during the preparatory steps preceding the direct immersion in liquid nitrogen, in order to overcome until now unsuccessful cryopreservation with this species. The exposure of shoot tips to 0.3-0.4 M sucrose leads to a high cell solute concentration. The treatment with plant vitrification solution (PVS2) alone, i.e., not followed by storage in liquid nitrogen, markedly affected shoot tip survival. After a 30 min exposure, regrowth percentage of shoot tips decreased from 94 % (control) to 57 %, and dropped to 15 % when the treatment was prolonged up to 60 min. After a 90 min exposure, no regrowth occurred. In addition, plantlets regenerated from shoot tips which underwent 60 min or more exposure to PVS2 showed signs of malformation. Microscope observations of shoot tips treated with 0.3 or 0.4 M sucrose and 30 min PVS2 showed the presence of cells starting to plasmolyze, localized in the area surrounding the apical meristem. A limited presence of starch grains in meristem and bract cells was also noted. However, the most conspicuous consequence of prolonged PVS2 treatment was convex plasmolysis. The phenomenon was dependent on the time of PVS2 exposure. Indeed, after a 30 min treatment, plasmolysis was minimal or absent, but it increased with longer exposure to PVS2 at 4 °C.

SUSCEPTIBILITY OF THOMPSON SEEDLESS GRAPEVINES RAISED ON DIFFERENT ROOTSTOCKS TO LEAF BLACKENING AND NECROSIS UNDER SALINE IRRIGATION

Saline irrigation water having high sodium (Na+) content leads to the development of black leaf disorder in the vines resulting in reduced productivity and death of permanent vine parts. The response of the vines raised on different rootstocks differs under such conditions. Investigation was carried out to study the susceptibility of grafted and own-rooted Thompson Seedless (Vitis vinifera L.) vines to black leaf disorder. Grapevines grafted on four different rootstocks viz. Dog Ridge (Vitis champini), Salt Creek (Vitis champini), B2-56 (Vitis berlandieri x Vitis rupestris) and 1613C [Vitis longii x (Vitis labrusca x Vitis riparia) x Vitis vinifera] were studied. The variations in nutrient content of various vine parts under saline irrigation in relation to leaf blackening and necrosis symptoms were also studied. Thompson Seedless vines grafted on Dog Ridge, Salt Creek and own rooted vines started developing leaf blackening and necrosis symptoms during the ripening stage. Vines exhibiting these symptoms contained significantly lower potassium (K+) and higher Na+ content in the leaf blades as compared to healthy vines. Vines grafted on Salt Creek showed most severe leaf blackening and necrosis symptoms and had the lowest K+ content in blades and petioles. Amongst the rootstocks, vines raised on B2-56 had the lowest Na+ concentration in leaf blades whereas those on Dog Ridge and Salt Creek accumulated Na+ in leaf blades to a toxic level. Significantly higher K+ content was found in vines raised on B2-56 and 1613C rootstocks in all the vine parts compared to other stock-scion combinations. Vines grafted on B2-56 and 1613C could maintain higher K+: Na+ ratio and tolerated saline irrigation better than other stock-scion combinations. All the rootstocks reduced chloride concentration significantly in the vine parts (petioles, blade and canes) compared to own rooted vines. Vines raised on their own roots and on B2-56 rootstock had significantly higher phosphorus (P) concentration in petiole, blade and canes compared to vines grafted on Dog Ridge and Salt Creek. Total biomass (petiole + blade + cane on dry weight basis) and yield was the highest in B2-56 rootstock and least in 1613C.

The volatile metabolome of grapevine roots: first insights into the metabolic response upon phylloxera attack.

Many plant species respond to herbivore attack by an increased formation of volatile organic compounds. In this preliminary study we analysed the volatile metabolome of grapevine roots [Teleki 5C (Vitis berlandieri Planch. × Vitis riparia Michx.)] with the aim to gain insight into the interaction between phylloxera (Daktulosphaira vitifoliae Fitch; Hemiptera: Phylloxeridae) and grapevine roots. In the first part of the study, headspace solid phase microextraction (HS-SPME) coupled to gas chromatography – mass spectrometry (GC–MS) was used to detect and identify volatile metabolites in uninfested and phylloxera-infested root tips of the grapevine rootstock Teleki 5C. Based on the comparison of deconvoluted mass spectra with spectra databases as well as experimentally derived retention indices with literature values, 38 metabolites were identified, which belong to the major classes of plant volatiles including C6-compounds, terpenes (including modified terpenes), aromatic compounds, alcohols and n-alkanes. Based on these identified metabolites, changes in root volatiles were investigated and resulted in metabolite profiles caused by phylloxera infestation. Our preliminary data indicate that defence related pathways such as the mevalonate and/or alternative isopentenyl pyrophosphate-, the lipoxygenase- (LOX) as well as the phenylpropanoid pathway are affected in root galls as a response to phylloxera attack.

Development of hybrid elution systems for efficient purification of stilbenoids using centrifugal partition chromatography coupled to mass spectrometry

The phytochemical study of the root extract of the stilbenoid-rich Vitis riparia×Vitis berlandieri grapevine was carried out by centrifugal partition chromatography (CPC). For this reason, we developed a new elution mode we named back-step, which allowed us to obtain cleaner fractions and a more efficient separation process when used in conjunction with a classical elution approach. Three hydroxystilbenes: (E)-resveratrol, (E)-ɛ-viniferin and (E)-vitisin C, with greater than 90% purity were thus obtained through such process, with minimal sample handling and purification steps. Online coupling of CPC to ESI mass spectrometry was used for optimization of the separation parameters and to facilitate the characterization of the stilbenoids. This study details the first phytochemical investigation of stilbenoids from the hybrid species together with a new elution mode able to widen the range of ARIZONA biphasic systems.

Salinity Responses of Grafted Grapevines: Effects of Scion and Rootstock Genotypes

Two-year-old Sultana and Mke vines (Vitis vinifera L.) grafted onto Rupestris du Lot (Vitis rupestris Scheele) and 110 R (Vitis berlandieri Vitis rupestris) were grown in soil, sand, sphagnum peat and farmyard manure mixture (2:1:1:1 v/v) irrigated with 0.3, 2.7 and 5.45 dS m-1 NaCl solutions for a period of 60 days. Biomass accumulation, relative chlorophyll content, leaf water potential, stomatal conductance and transpiration were significantly reduced by salinity in all graft combinations. The inhibiting effect of salinity on growth varied according to the characteristics of the scion genotype and the level of salinity. The results imply the predominance of the scion genotype in determining variation in the leaf-level physiological characteristics of grafted vines because stomatal conductance and transpiration were higher in Sultana than in Mke at the same leaf water potential and the level of salinity, regardless of the rootstock genotype. NaCl treatments did not affect leaf Cu concentration while inducing significant Na, K, Ca, N, P, Mg, Fe, Mn and Zn elevations in the leaves of all scion/rootstock combinations. While salinity caused an increase in N content and a decrease in K content in the roots of all graft combinations, it had no influence on Ca, P, Cu and Zn concentrations. In contrast to the Sultana grafted vines, NaCl salinity led to a decrease in Mg, Fe and Mn concentrations in the roots of Mke grafted vines. Nevertheless, ion accumulation in leaves and roots of grafted vines under saline or non-saline conditions was controlled predominantly by the genotype of the scion. In response to salinity, Sultana grafts accumulated higher amounts of ions in their leaves than in their roots. However, in Mke, higher ion concentrations were seen in the roots. Therefore, the increase in the transport of inorganic ions up to the leaves seems to be the major component of osmotic adjustment in salt-stressed Sultana vines.

Contrast in chloride exclusion between two grapevine genotypes and its variation in their hybrid progeny

Potted grapevines of 140 Ruggeri (Vitis berlandieri × Vitis rupestris), a good Cl− excluder, and K 51-40 (Vitis champinii × Vitis riparia ‘Gloire’), a poor Cl− excluder, and of a family obtained by crossing the two genotypes, were used to examine the inheritance of Cl− exclusion. Rooted leaves were then used to further investigate the mechanism for Cl− exclusion in 140 Ruggeri. In both a potting mix trial (plants watered with 50 mM Cl−) and a solution culture trial (plants grown in 25 mM Cl−), the variation in Cl− accumulation was continuous, indicating multiple rather than single gene control for Cl− exclusion between hybrids within the family. Upper limits of 42% and 35% of the phenotypic variation in Cl− concentration could be attributed to heritable sources in the potting mix and solution culture trials, respectively. Chloride transport in roots of rooted leaves of both genotypes appeared to be via the symplastic pathway, since addition of 8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS), an apoplastic tracer, revealed no obvious PTS fluorescence in the laminae of either genotype, despite significant accumulation of Cl− in laminae of K 51-40 during the PTS uptake period. There was no significant difference in either unidirectional 36Cl− flux (10 min) or 36Cl− uptake (3 h) into roots of rooted leaves exposed to 5, 10, or 25 mM Cl−. However, the percentage of 36Cl− transported to the lamina (3 h) was significantly lower in 140 Ruggeri than in K 51-40, supporting reduced Cl− loading into xylem and implicating the root stele in the Cl− exclusion mechanism.

Seasonal changes of whole root system conductance by a drought-tolerant grape root system

The role of root systems in drought tolerance is a subject of very limited information compared with above-ground responses. Adjustments to the ability of roots to supply water relative to shoot transpiration demand is proposed as a major means for woody perennial plants to tolerate drought, and is often expressed as changes in the ratios of leaf to root area (AL:AR). Seasonal root proliferation in a directed manner could increase the water supply function of roots independent of total root area (AR) and represents a mechanism whereby water supply to demand could be increased. To address this issue, seasonal root proliferation, stomatal conductance (gs) and whole root system hydraulic conductance (kr) were investigated for a drought-tolerant grape root system (Vitis berlandieri×V. rupestris cv. 1103P) and a non-drought-tolerant root system (Vitis riparia×V. rupestris cv. 101-14Mgt), upon which had been grafted the same drought-sensitive clone of Vitis vinifera cv. Merlot. Leaf water potentials (ψL) for Merlot grafted onto the 1103P root system (–0.91±0.02 MPa) were +0.15 MPa higher than Merlot on 101-14Mgt (–1.06±0.03 MPa) during spring, but dropped by approximately –0.4 MPa from spring to autumn, and were significantly lower by –0.15 MPa (–1.43±0.02 MPa) than for Merlot on 101-14Mgt (at –1.28±0.02 MPa). Surprisingly, gs of Merlot on the drought-tolerant root system (1103P) was less down-regulated and canopies maintained evaporative fluxes ranging from 35–20 mmol vine−1 s−1 during the diurnal peak from spring to autumn, respectively, three times greater than those measured for Merlot on the drought-sensitive rootstock 101-14Mgt. The drought-tolerant root system grew more roots at depth during the warm summer dry period, and the whole root system conductance (kr) increased from 0.004 to 0.009 kg MPa−1 s−1 during that same time period. The changes in kr could not be explained by xylem anatomy or conductivity changes of individual root segments. Thus, the manner in which drought tolerance was conveyed to the drought-sensitive clone appeared to arise from deep root proliferation during the hottest and driest part of the season, rather than through changes in xylem structure, xylem density or stomatal regulation. This information can be useful to growers on a site-specific basis in selecting rootstocks for grape clonal material (scions) grafted to them.

In vitro interaction studies between Glomus intraradices and Armillaria mellea in vines

An interaction study was performed with mycorrhizal plants of the grapevine rootstock Richter 110 (Vitis berlandieri Planch × Vitis rupestris Scheele) and the root pathogenic fungus Armillaria mellea (Vahl:Fr.) P. Kumm using an autotrophic in vitro culture system. Micropropagated plantlets were transferred to Petri plates with MSR medium lacking sugar and vitamins. Inocula of Glomus intraradices (BEG 72) and of Armillaria mellea obtained from a root organ culture and from a mycelium colony grown in malt agar respectively, were added to the plates according to each treatment: non-inoculated, inoculated with G. intraradices, inoculated with A. mellea, and dual-inoculated plants. There were ten replicates per treatment. Fourteen weeks later, the pathogen's mycelium occupied most of the surface/volume of the plate and had produced rhizomorphs. In dual inoculated plates, A. mellea's growth was not affected by the presence of G. intraradices, but the latter produced a lower number of spores and its extraradical phase showed granulation, vacuolation and tip swelling. The pathogen induced necrosis and growth decrease in the root. Glomus intraradices alleviated these symptoms and there were no differences in root biomass between non-inoculated plants and plants inoculated with both fungi. There were no symptoms of the disease in shoots and G. intraradices stimulated shoot growth both, although mycorrhizal colonization was lower when A. mellea was present. No direct antagonism or antibiosis against the pathogen was observed, thus the protective effect exerted by the symbiotic fungus in grapevines must be indirect, mediated through the host plant physiology.

Susceptibility of grapevine viruses to thermotherapy on in vitro collection of Kober 5BB

A number of virus elimination protocols have been developed to obtain healthy plants but their success rates have been variable due to the strong virus–host relationship, treatment performances and experimental condition. An in vitro collection of plants of Vitis berlandieri × Vitis riparia Kober 5BB single-infected by Grapevine vitivirus A (GVA), Grapevine fanleaf nepovirus (GFLV), Grapevine fleck maculavirus (GFkV), Grapevine leafroll ampelovirus 1 (GLRaV-1) and Grapevine leafroll ampelovirus 3 (GLRaV-3) was established to carry out thermotherapy treatments. The experimental system allowed to control the effects of host genotype by using the same genetic background, and the environmental condition such as relative humidity, soil and temperature conditions. Trials were conducted in a growth chamber set at 37 ± 0.5 °C for 48 days using in vitro cultures for all infections. ELISA and RT-PCR were used to check sanitary conditions. Results showed complete eradication of GFLV and no effect against GFkV. Different sanitation rates were obtained for other phloematic viruses: 70.2% for GVA, 25.1% for GFLaV-1 and 24.7% for GLRaV-3. The results on phloematic viruses showed a gradient of elimination efficiency which could not be completely explained with the location of viral particles that seems to be only one of the several factors affecting virus susceptibility to heat stress. Moreover, the advantages of this experimental system provide the opportunity to obtain a greater understanding of the thermotherapeutic effects on phloematic viruses.

Effect of Medicago sativa ferritin gene on stress tolerance in transgenic grapevine

The effect of Medicago sativa (alfalfa) ferritin gene (MsFer) on abiotic stress tolerance was tested using transgenic Vitis berlandieri × Vitis rupestris cv. ‘Richter 110’ grapevine rootstock lines. Leaf discs from transgenic plants maintained higher photosynthetic activity after NaCl, tert-butyl-hydroperoxide (t-BHP) or paraquat treatment than control ones. These results indicate that the increased production of ferritin significantly improved abiotic stress tolerance in transgenic grapevine plants.

Differential Growth of Mycorrhizal Field-Inoculated Grapevine Rootstocks in Two Replant Soils

The mycorrhizal field inoculation of 161-49 Couderc (<i>Vitis riparia</i> Michx. x <i>Vitis berlandieri</i> Planch.) and 140 Ruggeri (<i>Vitis rupestris</i> L. x <i>V. berlandieri</i>) grapevine rootstock plants grafted with Cabernet Sauvignon was evaluated in two replant soils of a Mediterranean production area in northeastern Spain. The first soil (vineyard 1) had been under tillage for 10 years. The second soil (vineyard 2) had been cultivated for one year before setting up the new plantation and was infested by the white root rot fungus <i>Armillaria mellea</i> (Vahl:Fr.) P. Kumm. The number of infective mycorrhizal propagules estimated in the soil before planting was 1 per 100 mL soil in vineyard 1 and none in vineyard 2. Half of the vines for each rootstock were inoculated with the arbuscular mycorrhizal fungus (AMF) <i>Glomus intraradices</i> Schenck and Smith (BEG 72). In vineyard 2, lacking mycorrhizal propagules and infested with <i>A. mellea</i>, soil was also inoculated before planting with <i>Thymus vulgaris</i> L. and <i>Lavandula officinalis</i> Mill. plants colonized by <i>G. intraradices</i> in two rows. At the end of the second growing season, 19 months after planting, the direct field inoculation showed a beneficial effect on the growth of 140 Ruggeri vines in vineyard 1. The total biomass of inoculated 161-49 Couderc vines increased in vineyard 2 but not in vineyard 1. Precropping with mycorrhizal carrier plants increased the mycorrhizal potential of the replant soil. Grapevine response to AMF inoculation was influenced by the intrinsic characteristics of the vineyard soil and by the rootstock used and the time after planting.

Grapevine cell early activation of specific responses to DIMEB, a resveratrol elicitor

BackgroundIn response to pathogen attack, grapevine synthesizes phytoalexins belonging to the family of stilbenes. Grapevine cell cultures represent a good model system for studying the basic mechanisms of plant response to biotic and abiotic elicitors. Among these, modified β-cyclodextrins seem to act as true elicitors inducing strong production of the stilbene resveratrol.ResultsThe transcriptome changes of Vitis riparia × Vitis berlandieri grapevine cells in response to the modified β-cyclodextrin, DIMEB, were analyzed 2 and 6 h after treatment using a suppression subtractive hybridization experiment and a microarray analysis respectively. At both time points, we identified a specific set of induced genes belonging to the general phenylpropanoid metabolism, including stilbenes and hydroxycinnamates, and to defence proteins such as PR proteins and chitinases. At 6 h we also observed a down-regulation of the genes involved in cell division and cell-wall loosening.ConclusionsWe report the first large-scale study of the molecular effects of DIMEB, a resveratrol inducer, on grapevine cell cultures. This molecule seems to mimic a defence elicitor which enhances the physical barriers of the cell, stops cell division and induces phytoalexin synthesis.

COLLECTING VITIS BERLANDIERI FROM NATIVE HABITAT SITES

Lime content is the major difference between Northern American and European vine growing soils. This resulted in significant difficulties in the development of rootstocks for European conditions at the end of the 19th and the beginning of the 20th century. It was only the introduction of Vitis berlandieri as a breeding partner that led to lime-tolerant rootstocks. Despite the importance of V. berlandieri for European viticulture, only a few accessions have so far been exploited for breeding rootstocks. In most cases, these vines were only used because they were available in Europe at that time. The genetic variability of the species is certainly much larger. To preserve and evaluate genetic diversity in V. berlandieri, grape berries from 86 individual vines were collected in September 2005 from a large range of natural stands in Texas, United States. The collection locations were found in 13 counties and across a distribution from N 31° 23' to N 29° 43' and W 100° 2' to W 97° 26'. Vines will be planted in germplasm collections and evaluated for their rooting and grafting ability, their lime tolerance, and other viticultural features. Superior types will then be utilized for rootstock breeding programs.