Canopy Architecture and Sun Exposure Influence Berry Cluster-Water Relations in the Grapevine Variety Muscat of Alexandria.

Climate-change-related increases in the frequency and intensity of heatwaves affect viticulture, leading to losses in yield and grape quality. We assessed whether canopy-architecture manipulation mitigates the effects of summer stress in a Mediterranean vineyard. The Vitis vinifera L variety Muscat of Alexandria plants were monitored during 2019-2020. Two canopy shoot-positioning treatments were applied: vertical shoot positioning (VSP) and modulated shoot positioning (MSP). In MSP, the west-side upper foliage was released to promote partial shoot leaning, shading the clusters. Clusters were sampled at pea size (PS), veraison (VER), and full maturation (FM). Measurements included rachis anatomy and hydraulic conductance (Kh) and aquaporins (AQP) and stress-related genes expression in cluster tissues. The results show significant seasonal and interannual differences in Kh and vascular anatomy. At VER, the Kh of the rachis and rachis+pedicel and the xylem diameter decreased but were unaffected by treatments. The phloem-xylem ratio was either increased (2019) or reduced (2020) in MSP compared to VSP. Most AQPs were down-regulated at FM in pedicels and up-regulated at VER in pulp. A potential maturation shift in MSP was observed and confirmed by the up-regulation of several stress-related genes in all tissues. The study pinpoints the role of canopy architecture in berry-water relations and stress response during ripening.

Modulation of the Berry Skin Transcriptome of cv. Tempranillo Induced by Water Stress Levels.

Climate change in the Mediterranean area is making summers warmer and dryer. Grapevine (Vitis vinifera L.) is mostly important for wine production in Mediterranean countries, and the variety Tempranillo is one of the most cultivated in Spain and Portugal. Drought decreases yield and quality and causes important economic losses. As full irrigation has negative effects on quality and water is scarce in this region, deficit irrigation is often applied. In this research, we studied the effects of two deficit irrigation treatments, Sustained Deficit Irrigation (SDI) and Regulated Deficit Irrigation (RDI), on the transcriptome of grape berries at full maturation, through RNAseq. The expression of differentially regulated genes (DEGs) was also monitored through RT-qPCR along berry development. Most transcripts were regulated by water stress, with a similar distribution of up- and down-regulated transcripts within functional categories (FC). Primary metabolism was the more severely affected FC under water stress, followed by signaling and transport. Almost all DEGs monitored were significantly up-regulated by severe water stress at veraison. The modulation of an auxin response repression factor, AUX22D, by water stress indicates a role of this gene in the response to drought. Further, the expression of WRKY40, a TF that regulates anthocyanin biosynthesis, may be responsible for changes in grape quality under severe water stress.

Developmental Regulation of Transcription in Touriga Nacional Berries under Deficit Irrigation.

Grapevine (Vitis vinifera L.) is one of the most economically important crops worldwide, especially due to the economic relevance of wine production. Abiotic stress, such as drought, may contribute to low yield, shifts in quality, and important economic loss. The predicted climate change phenomena point to warmer and dryer Mediterranean environmental conditions; as such, it is paramount to study the effects of abiotic stress on grapevine performance. Deficit irrigation systems are applied to optimize water use efficiency without compromising berry quality. In this research, the effect of two deficit irrigation strategies, sustained deficit irrigation (SDI) and regulated deficit irrigation (RDI), in the grape berry were assessed. The effects of different levels of drought were monitored in Touriga Nacional at key stages of berry development (pea size, véraison, and full maturation) through RNA-Seq transcriptome analysis and by specific differentially expressed genes (DEGs) monitoring through RT-qPCR. Handy datasets were obtained by bioinformatics analysis of raw RNA-Seq results. The dominant proportion of transcripts was mostly regulated by development, with véraison showing more upregulated transcripts. Results showed that primary metabolism is the functional category more severely affected under water stress. Almost all DEGs selected for RT-qPCR were significantly upregulated in full maturation and showed the highest variability at véraison and the lowest gene expression values in the pea size stage.

Mycorrhizal Inoculation Differentially Affects Grapevine's Performance in Copper Contaminated and Non-contaminated Soils.

Plant inoculation with arbuscular mycorrhizal fungi (AMF) is increasingly employed to enhance productivity and sustainability in agricultural ecosystems. In the present study, the potential benefits of AMF inoculation on young grapevines replanted in pots containing vineyard soil with high Cu concentration were evaluated. For this purpose, one-year-old cv. Touriga Nacional grapevines grafted onto 1103P rootstocks were further inoculated with Rhizoglomus irregulare or Funneliformis mosseae, or left non-inoculated, and maintained in a sterilized substrate under greenhouse conditions for three months. After this time, half of the plants were transplanted to containers filled with an Arenosol from a vineyard which had been artificially contaminated or not with 300 mg kg-1 of Cu. At the end of the growing season, soil nutrient concentration, soil dehydrogenase activity and mycorrhizal colonization rate were analyzed. Grapevine performance was assessed by measuring several vegetative growth and physiological parameters as well as nutrient concentrations in leaves and roots. In the non-contaminated soil, R. irregulare- and F. mosseae-inoculated plants had significantly greater root biomass than the non-inoculated ones. However, the opposite effect was observed in the Cu-contaminated soil, where non-inoculated plants performed better regarding shoot and root development. Concerning nutrient levels, an increase in Cu, Mg and Mn concentrations was observed in the roots of plants growing in the contaminated soil, although only Mn was translocated to leaves. This led to a large increase in leaf Mn concentrations, which was significantly higher in non-inoculated and F. mosseae- inoculated plants than in the R. irregulare- inoculated ones. Copper contamination induced a general decrease in leaf N, P and Fe concentrations as well as chlorosis symptoms. The largest decrease in N and P was observed in F. mosseae- inoculated plants, with 73 and 31.2%, respectively. However, these plants were the ones with the least decrease in Fe concentration (10% vs. almost 30% in the other two inoculation treatments). In conclusion, this study indicates that soil Cu levels can modify the outcome of AMF inoculations in young grapevines, disclosing new AMF-plant associations potentially relevant in vineyards with a tradition of Cu-based fungicide application.

Grape Ripening Is Regulated by Deficit Irrigation/Elevated Temperatures According to Cluster Position in the Canopy.

The impact of water deficit on berry quality has been extensively investigated during the last decades. Nonetheless, there is a scarcity of knowledge on the performance of varieties exposed to a combination of high temperatures/water stress during the growing season and under vineyard conditions. The objective of this research was to investigate the effects of two irrigation regimes, sustained deficit irrigation (SDI, 30% ETc) and regulated deficit irrigation (RDI, 15% ETc) and of two cluster positions within the canopy (east- and west-exposed sides) on berry ripening in red Aragonez (Tempranillo) grapevines. The study was undertaken for two successive years in a commercial vineyard in South Portugal, monitoring the following parameters: pre-dawn leaf water potential, berry temperature, sugars, polyphenols, abscisic acid (ABA) and related metabolites. Additionally, expression patterns for different transcripts encoding for enzymes responsible for anthocyanin and ABA biosynthesis (VviUFGT, VvNCED1, VvßG1, VviHyd1, VviHyd2) were analyzed. In both years anthocyanin concentration was lower in RDI at the west side (RDIW- the hottest one) from véraison onwards, suggesting that the most severe water stress conditions exacerbated the negative impact of high temperature on anthocyanin. The down-regulation of VviUFGT expression revealed a repression of the anthocyanin synthesis in berries of RDIW, at early stages of berry ripening. At full-maturation, anthocyanin degradation products were detected, being highest at RDIW. This suggests that the negative impact of water stress and high temperature on anthocyanins results from the repression of biosynthesis at the onset of ripening and from degradation at later stages. On the other hand, berries grown under SDI displayed a higher content in phenolics than those under RDI, pointing out for the attenuation of the negative temperature effects under SDI. Irrigation regime and berry position had small effect on free-ABA concentration. However, ABA catabolism/conjugation process and ABA biosynthetic pathway were affected by water and heat stresses. This indicates the role of ABA-GE and catabolites in berry ABA homeostasis under abiotic stresses. Principal component analysis (PCA) showed that the strongest influence in berry ripening is the deficit irrigation regime, while temperature is an important variable determining the improvement or impairment of berry quality by the deficit irrigation regime. In summary, this work shows the interaction between irrigation regime and high temperature on the control of berry ripening.

Performance of an adipokine pathway-based multilocus genetic risk score for prostate cancer risk prediction.

Few biomarkers are available to predict prostate cancer risk. Single nucleotide polymorphisms (SNPs) tend to have weak individual effects but, in combination, they have stronger predictive value. Adipokine pathways have been implicated in the pathogenesis. We used a candidate pathway approach to investigate 29 functional SNPs in key genes from relevant adipokine pathways in a sample of 1006 men eligible for prostate biopsy. We used stepwise multivariate logistic regression and bootstrapping to develop a multilocus genetic risk score by weighting each risk SNP empirically based on its association with disease. Seven common functional polymorphisms were associated with overall and high-grade prostate cancer (Gleason≥7), whereas three variants were associated with high metastatic-risk prostate cancer (PSA≥20 ng/mL and/or Gleason≥8). The addition of genetic variants to age and PSA improved the predictive accuracy for overall and high-grade prostate cancer, using either the area under the receiver-operating characteristics curves (P<0.02), the net reclassification improvement (P<0.001) and integrated discrimination improvement (P<0.001) measures. These results suggest that functional polymorphisms in adipokine pathways may act individually and cumulatively to affect risk and severity of prostate cancer, supporting the influence of adipokine pathways in the pathogenesis of prostate cancer. Use of such adipokine multilocus genetic risk score can enhance the predictive value of PSA and age in estimating absolute risk, which supports further evaluation of its clinical significance.

Tumor cell-educated periprostatic adipose tissue acquires an aggressive cancer-promoting secretory profile.

BACKGROUND/AIMS: The microenvironment produces important factors that are crucial to prostate cancer (PCa) progression. However, the extent to which the cancer cells stimulate periprostatic adipose tissue (PPAT) to produce these proteins is largely unknown. Our purpose was to determine whether PCa cell-derived factors influence PPAT metabolic activity. METHODS: Primary cultures of human PPAT samples from PCa patients (adipose tissue organotypic explants and primary stromal vascular fraction, SVF) were stimulated with conditioned medium (CM) collected from prostate carcinoma (PC3) cells. Cultures without CM were used as control. We used multiplex analysis and ELISA for protein quantification, qPCR to determine mitochondrial DNA (mtDNA) copy number and zymography for matrix metalloproteinase activity, in order to evaluate the response of adipose tissue explants and SVFs to PC3 CM. RESULTS: Stimulation of PPAT explants with PCa PC3 CM induced adipokines associated with cancer progression (osteopontin, tumoral necrosis factor alpha and interleukin-6) and reduced the expression of the protective adipokine adiponectin. Notably, osteopontin protein expression was 13-fold upregulated. Matrix metalloproteinase 9 activity and mitochondrial DNA copy number were higher after stimulation with cancer CM. Stromovascular cells from PPAT in culture were not influenced by tumor-derived factors. CONCLUSION: The modulation of adipokine expression by tumor CM indicates the pervasive extent to which tumor cells command PPAT to produce factors favorable to their aggressiveness.

Grapevine varieties exhibiting differences in stomatal response to water deficit.

Knowledge on variety traits and physiological responses to stress is still scarce in Vitis vinifera L., limiting the optimisation of irrigation and breeding for high water use efficiency. We have characterised five grapevine varieties using thermal imaging, leaf gas exchange, leaf morphology and carbon isotope composition. Plants of the varieties Aragonez, Trincadeira, Cabernet Sauvignon, Syrah and Touriga Nacional were grown in field conditions. Two experiments were performed. In Experiment I (2006), vines of Aragonez and Trincadeira were either well irrigated (WI, 80% ETc), non-irrigated but rain fed (NI) or subjected to regulated deficit irrigation (RDI, 40% ETc) and studied along the summer season. In Experiment II (2006 and 2007), vines of the five varieties were subjected to RDI (30-40% ETc) and studied at veraison. In Experiment I, leaf temperature (Tleaf) correlated negatively with stomatal conductance (gs) and leaf water potential (Ψpd). The inverse relationship between gs and Tleaf was highly significant in the afternoon. In Experiment II, the different genotypes showed different Tleaf for similar Ψpd. Stomatal density did not correlate with gs suggesting that varieties have different stomatal control. Our results show that combined measurements of canopy temperature and Ψpd can aid in better understanding of stomatal regulation in different grapevine varieties. Such variation in stomatal regulation should be taken into account in determining irrigation strategies.

Hydraulic and chemical signalling in the regulation of stomatal conductance and plant water use in field grapevines growing under deficit irrigation.

Effects of irrigation strategies on stomata and plant water use were studied in field-grown grapevines (Vitis vinifera L.). We assessed the importance of root-derived chemical signals vs. hydraulic signalling in stomatal regulation. The experiment included two treatments with the same water added to the soil (50% ETc) applied either to the whole root system (DI) or to half of the roots, alternating irrigation side every 15 days (PRD). Well-watered plants (FI) (100% ETc) and non-irrigated grapevines (NI) were also studied. Partial stomata closure occurred in both PRD and DI plants. [ABA] of xylem sap remained constant during the day and was maintained throughout the season, with higher values in NI plants. Xylem sap pH was not affected by soil water availability. A positive correlation between ψpd and maximum g s was found, indicating that grapevine stomata strongly respond to plant water status. In contrast, ABA did not explain stomatal control at veraison. At mid-ripening g s was significantly correlated with ABA, apparently interacting with the rise in xylem sap pH. Therefore, our data suggest that hydraulic feedback and feed-forward root-to-shoot chemical signalling mechanisms might be involved in the control of stomata in response to decreased soil water availability, hydraulic signals playing the dominant role.

Impact of deficit irrigation on water use efficiency and carbon isotope composition (delta13C) of field-grown grapevines under Mediterranean climate.

The objective of this study was to evaluate the effect of deficit irrigation on intrinsic water use efficiency (A/g(s)) and carbon isotope composition (delta13C) of two grapevine cultivars (Moscatel and Castelão), growing in a commercial vineyard in SW Portugal. The study was done in two consecutive years (2001 and 2002). The treatments were full irrigation (FI), corresponding to 100% of crop evapotranspiration (ETc), rain-fed (no irrigation, NI), and two types of deficit irrigation (50% ETc): (i) by supplying the water either to one side of the root system or to the other, which is partial rootzone drying (PRD), or (ii) dividing the same amount of water by the two sides of the root system, the normal deficit irrigation (DI). The water supplied to the PRD treatment alternated sides approximately every 15 d. The values of predawn leaf water potential (Psi(pd)) and the cumulative integral of Psi(pd) (S(Psi)) during the season were lower in 2001 than in the 2002 growing season. Whereas differences in Psi(pd) and S(Psi) between PRD and DI were not significantly different in 2001, in 2002 (a dryer year) both cultivars showed lower values of S(Psi) in the PRD treatment as compared with the DI treatment. This suggests that partial rootzone drying may have a positive effect on water use under dryer conditions, either as a result of better stomatal control and/or reduced vigour. The effects of the water treatments on delta13C were more pronounced in whole grape berries and pulp than in leaves. The delta13C of pulp showed the best correlation with intrinsic water use efficiency (A/g(s)) as well as with S(Psi). In spite of the better water status observed in PRD compared with DI in the two cultivars in 2002, no statistical differences between the two treatments were observed in A/g(s) and delta13C. On the other hand, they showed a higher delta13C compared with FI. In conclusion, it is apparent that the response to deficit irrigation varies with the environmental conditions of the particular year, the driest conditions exacerbating the differences among treatments. The highest values of delta13C found in the pulp of NI vines in Castelão compared with Moscatel suggest different sensitivities to water deficits in the two cultivars, as was empirically observed.

Partial rootzone drying: regulation of stomatal aperture and carbon assimilation in field-grown grapevines (Vitis vinifera cv. Moscatel).

The effects of 'partial rootzone drying' (PRD) irrigation compared with other irrigation systems, namely non-irrigated (NI), full irrigation (FI) and deficit irrigation (DI), on stomatal conductance and carbon assimilation were evaluated in field-grown grapevines (Vitis vinifera L. cv. Moscatel). At the end of the growing season, pre-dawn leaf water potential was highest in FI (-0.18 ± 0.01 MPa; mean ± s.e.), intermediate in PRD (-0.30± 0.01 MPa) and DI (-0.36 ± 0.02 MPa), and lowest in NI vines (-0.64 ± 0.03 MPa). Stomatal conductance measured under controlled conditions of light and temperature was reduced in NI (ca 60%) and PRD (ca 30%) vines compared with DI and FI vines. Under ambient conditions, NI vines had lower rates of stomatal conductance (ca26%), net CO2 assimilation (ca 28%) and light-adapted PSII quantum yields (ca 47%) than PRD, DI and FI vines. No significant differences were found among the three irrigated treatments. Both maximum electron transport rate (Jmax; ca 30%) and triose-phosphate utilization rates (TPU; ca 20%) were significantly lower in NI and PRD vines than in DI and FI vines. Carbon isotope composition (δ13C) of grape berries was highest in NI vines (-24.3), followed by PRD (-25.4) and DI (-25.8) and lowest in FI (-26.4) vines, suggesting a long-term increase in the efficiency of leaf gas exchange in NI compared with PRD, DI and FI vines. Sap-flow data and estimates of relative stomatal limitation are in accordance with the observed stomatal closure in PRD vines. In this study, we show that PRD irrigation was able to maintain a vine water status closed to FI, but with double water use efficiency, which was due to a reduction of stomatal conductance with no significant decrease in carbon assimilation.

Partial rootzone drying: effects on growth and fruit quality of field-grown grapevines (Vitis vinifera).

A study to assess the effects of the Partial Rootzone Drying (PRD) irrigation strategy in comparison to other irrigation systems was carried out in southern Portugal in two field-grown grapevines varieties, Moscatel and Castelão. We addressed the question of whether by regulating growth and plant water use, the PRD system would enable an equilibrated vegetative development, leading to a favourable capture of solar radiation for photoassimilate production and, at the same time to provide an optimum environment for fruit maturation. Three irrigation schemes were applied in addition to the non-irrigated (NI) vines: partial root drying (PRD), 50% of crop evapotranspiration (ETc), supplied to only one side of the root system while the other one was allowed to dry, alternating sides every 15 days; deficit irrigated (DI), 50% ETc supplied, half to each side of the root system and full irrigated (FI, 100% ETc). During the whole season FI plants of both varieties exhibited a high leaf predawn water potential (ψpd , ca-0.2MPa) while a progressive decline was observed in NI plants, reaching ψpd values near -0.7 MPa at the end of August. PRD and DI presented intermediate values. PRD vines exhibited a stronger control over vegetative growth as compared with DI and FI plants. This was expressed by lower values of total leaf area at harvest, leaf layer number, canopy wideness and water shoots number, allowing a higher light interception at the cluster zone that induced an improvement in some berry quality characteristics. Watering had no significant effects on sugar accumulation in the berries but led to a favourable increase in the must titratable acidity, mainly in Castelão. Whereas in DI and FI treatments berry skin anthocyanins and phenols content were always lower than in NI, in PRD there was either no reduction or the reduction was much lower than in the other irrigation treatments. Water use efficiency (WUE) was increased by about 80% in PRD and DI when compared with FI, as a result of almost similar yields in the three treatments. Yield gains of irrigated plants in relation to NI were modest, explained by the rainy spring in both years.