The pit membrane (PM) is a primary cell wall barrier that separates adjacent xylem water conduits, limiting the spread of xylem-localized pathogens and air embolisms from one conduit to the next. This paper provides a characterization of the size of the pores in the PMs of grapevine (Vitis vinifera). The PM porosity (PMP) of stems infected with the bacterium Xylella fastidiosa was compared with the PMP of healthy stems. Stems were infused with pressurized water and flow rates were determined; gold particles of known size were introduced with the water to assist in determining the size of PM pores. The effect of introducing trans-1,2-diaminocyclohexane-N,N,N´,N´-tetraacetic acid (CDTA), oligogalacturonides, and polygalacturonic acid into stems on water flux via the xylem was also measured. The possibility that cell wall-degrading enzymes could alter the pore sizes, thus facilitating the ability of X. fastidiosa to cross the PMs, was tested. Two cell wall-degrading enzymes likely to be produced by X. fastidiosa (polygalactuoronase and endo-1,4-b-glucanase) were infused into stems, and particle passage tests were performed to check for changes in PMP. Scanning electron microscopy of control and enzyme-infused stem segments revealed that the combination of enzymes opened holes in PMs, probably explaining enzyme impacts on PMP and how a small X. fastidiosa population, introduced into grapevines by insect vectors, can multiply and spread throughout the vine and cause Pierce's disease.

It is conventionally thought that multiplication of the xylem-limited bacterium Xylella fastidiosa (Xf) within xylem vessels is the sole factor responsible for the blockage of water movement in grapevines (Vitis vinifera) affected by Pierce's disease. However, results from our studies have provided substantial support for the idea that vessel obstructions, and likely other aspects of the Pierce's disease syndrome, result from the grapevine's active responses to the presence of Xf, rather than to the direct action of the bacterium. The use of magnetic resonance imaging (MRI) to observe the distribution of water within the xylem has allowed us to follow nondestructively the development of vascular system obstructions subsequent to inoculation of grapevines with Xf. Because we have hypothesized a role for ethylene produced in vines following infection, the impact of vine ethylene exposure on obstruction development was also followed using MRI. In both infected and ethylene-exposed plants, MRI shows that an important proportion of the xylem vessels become progressively air embolized after the treatments. The loss of xylem water-transporting function, assessed by MRI, has been also correlated with a decrease in stem-specific hydraulic conductivity (K  S) and the presence of tyloses in the lumens of obstructed water conduits. We have observed that the ethylene production of leaves from infected grapevines is greater than that from healthy vines and, therefore, propose that ethylene may be involved in a series of cellular events that coordinates the vine's response to the pathogen.

Anthocyanins are secondary metabolites synthesized in grape berry skins via the phenylpropanoid pathway, with functions ranging from skin coloration to protection against pathogens or UV light. Accumulation of these compounds is highly variable depending on genetics, environmental factors and viticultural practices. Besides their biological functions, anthocyanins improve wine quality, as a high anthocyanin content in berries has a positive impact on the color, total phenolic concentration and, ultimately, the price of wine. The present work studies the effect of the pre-veraison application of pectin derived oligosaccharides (PDO) on the synthesis and accumulation of these compounds, and associates the changes observed with the expression of key genes in the phenylpropanoid pathways. To this end, pre-veraison Cabernet Sauvignon bunches were treated with PDO to subsequently determine total anthocyanin content, the anthocyanin profile (by HPLC-DAD) and gene expression (by qRT-PCR), using Ethrel and water treatments for comparison. The results show that PDO were as efficient as Ethrel in generating a significant rise in total anthocyanin content at 30 days after treatment (dat), compared with water treatments (1.32, 1.48 and 1.02 mg e.Mv-3G/g FW respectively) without any undesirable effect on berry size, soluble solids, tartaric acid concentration or pH. In addition, a significant alteration in the anthocyanin profile was observed. Specifically, a significant increase in the relative concentration of malvidin was observed for both PDO and Ethrel treatments, compared with water controls (52.8; 55.0 and 48.3%, respectively), with a significant rise in tri-hydroxylated forms and a fall in di-hydroxylated anthocyanins. The results of gene expression analyses suggest that the increment in total anthocyanin content is related to a short term increase in phenylalanine ammonia-lyase (PAL) expression, mediated by a decrease in MYB4A expression. A longer term increase in UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT) expression, probably mediated by a rise in MYBA1 was also observed. Regarding the anthocyanin profile, despite the increase observed in MYB5A expression in PDO and Ethrel treatments, no changes in flavonoid 3'-hydroxylase (F-3'-H); flavonoid 3'5'-hydroxylase (F-3'5'-H) or O-methyltransferase (OMT) could be related with the profile modifications described. Overall, this study highlights that application of PDO is a novel means of altering specific grape berry anthocyanins, and could be a means of positively influencing wine quality without the addition of agrochemicals.

Background and Aims Tyloses and embolisms have been reported to impair water transport during the development of Pierce's disease (PD), caused by the xylem‐dwelling pathogen Xylella fastidiosa. This work investigates the relative importance of these xylem conduit obstructions in stems of inoculated Vitis vinifera plants. Methods and Results After 18 weeks in a greenhouse, internodes from control and needle‐inoculated Chardonnay vines were assessed for embolisms using non‐destructive magnetic resonance imaging (MRI). Tyloses were identified by subsequent destructive histology and light microscopy examination. Embolism of individual or small sectors of vessels was detected by MRI analysis of a control shoot, usually in the proximity of the pith; virtually no tyloses were observed in the corresponding histological sections. From a symptomatic (inoculated) shoot, MRIs revealed large areas of embolised vessels extending radially towards the epidermis in most of the internodes; several individual or groups of two to three vessels appeared to be occluded with tyloses. Conclusions In all assessed internodes, MRI‐detected embolisms were found more frequently than tyloses, suggesting that over the course of PD embolism formation precedes tylose development. Significance of the Study Embolisms appear to be the first and predominant type of occlusion in stems during early PD progression; nevertheless, their importance for explaining water transport impairment has been overlooked in most studies.

Color and other quality parameters of “Redglobe” grape (Vitis vinifera L.) berries were evaluated after treatment with brassinosteroid (BR) analogs. Three BRs analogs (24-epibrassinolide, Triol, or Lactone) were applied at three concentrations (0.0, 0.4, or 0.8 mg⋅L-1), at the onset of veraison. A commercial formulation (B-2000®) was also applied, at a recommended rate of 0.06 mg⋅L-1. The tested BR analogs were effective improving berry color (evaluated as color index for red grapes, CIRG), increasing the levels of soluble solids and anthocyanins, and changing the types of anthocyanins present without altering other quality and yield parameters. The effects of BR analogs on color enhancement could be explained by an increase in soluble solids content and/or anthocyanin content. Treatment with 24-epibrassinolide (at 0.4 mg⋅L-1) or the commercial formulation tended to favor the production of dihydroxylated anthocyanins, which are responsible for the red and pink colors of grape berries. Results indicate that the use of BRs constitutes a potential tool in the production of table grapes. This is the first report of this enhancement effect in a productive context.

Grapevine rupestris stem pitting associated virus (GRSPaV) is one of the most widely distributed viruses; even so, little is known about its effect on Vitis vinifera. To provide new insights, the effects of single and mixed GRSPaV infections on the V. vinifera cultivar "Cabernet Sauvignon" were studied by evaluating growth parameters, such as measurements of the total plant length, the number and distance of internodes and the number of leaves per shoot. In addition, parameters relating to gas exchange, i.e., the stomatal conductance, net photosynthetic rate, internal CO 2 concentration and leaf transpiration, were also assessed. All the measurements were performed in one-and two-yearold plants with a single GRSPaV infection or mixed infections of GRSPaV and Grapevine fanleaf virus (GFLV). The results show that the plant phytosanitary status did not significantly alter the growth and gas exchange parameters in oneyear-old plants. However, in two-year-old plants, single GRSPaV infections increased shoot elongation, which was accompanied by the overexpression of genes associated with the gibberellic acid response pathway. The gas exchange parameters of these plants were negatively affected, despite exhibiting higher LHCII gene expression. Plants with mixed infections did not have modified growth parameters, although they presented a greater reduction in the primary photosynthetic parameters evaluated with no change in LHCII expression. The results presented here confirm the co-evolution hypothesis for V. vinifera and GRSPaV during the early stages of plant development, and they provide new evidence about the effects of GRSPaV and GFLV co-infections on the "Cabernet Sauvignon" cultivar.

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In the near future, stomatal behaviour will be crucial to counteract conditions arising from climate change. Grapevine varieties are classified as either isohydric or anisohydric, depending on the sensitivity of stomata to water deficit and on their water potential homeostasis. However, the great variability observed in different studies indicates that a continuum exists in the range of stomatal sensitivity to water stress. Thus, more knowledge about the hydric behaviour and the gas exchange of isohydric and anisohydric grapevine varieties under different water conditions could lead to the development of irrigation strategies oriented at improving water-use efficiency, yield and berry composition. In this study, research was conducted in order to characterise the stomatal regulation of four different Vitis vinifera L. varieties, namely Pinot noir, Sauvignon blanc, Chardonnay and Merlot, according to soil water status. Measurements of leaf gas exchange, together with measurements of stem water potential (Ψs) and leaf water potential (Ψl), were taken during two seasons. Under conditions of water stress, Chardonnay and Merlot reached a minimum Ψs of-1.67 and-1.68 MPa respectively, and higher levels of water-use efficiency (A N/ g s), of 62.3 and 69.7 μmol CO 2/ mol H 2 O respectively. In Sauvignon blanc and Pinot noir, the minimum Ψs was-1.26 and-1.40 MPa respectively, with lower levels of A N/ g s (53.1 and 50.5 μmol CO 2/ mol H 2 O, respectively). Under conditions of water stress (Ψl <-0.9 MPa and Ψs <-0.6 MPa), all varieties had a significantly increased A N/ g s 1 , despite a significant reduction in gas exchange. Therefore, the hydric behaviour and gas exchange observed in this study suggest that Chardonnay and Merlot could be characterised as anisohydric varieties, as they present less sensitive stomatal control, while Pinot noir can be classified as a near-anisohydric variety and Sauvignon blanc as an isohydric variety. New investigations should consider other characteristics of the varieties to classify them better.