Water deficit before veraison is crucial in regulating berry VOCs concentration in Sangiovese grapevines.

The impact of water deficit on volatile organic compounds (VOCs) responsible for grape aroma remains quite unclear. The aim of this study was to evaluate the effect of different timing and intensity of water deficit on berry VOCs and on their biosynthetic pathways. Fully irrigated control vines were compared with the following treatments: i) two different levels of water deficit from berry pea-size through veraison, ii) one level of water deficit during the lag-phase, and iii) two different levels of water deficit from veraison through harvest. At harvest, total VOC concentrations were higher in berries of water stressed vines from berry pea size through veraison or during the lag phase, whereas post-veraison water deficit determined similar concentrations as control. This pattern was even more pronounced for the glycosylated fraction and was also observed for single compounds, mainly monoterpenes and C13-norisoprenoids. On the other hand, free VOCs were higher in berries from lag phase or post-veraison stressed vines. The significant glycosylated and free VOCs increment measured after the short water stress limited to the lag phase highlight the pivotal role played by this stage in berry aroma compound biosynthesis modulation. The severity of water stress before veraison was also important, since glycosylated VOCs showed a positive correlation with the pre-veraison daily water stress integral. The RNA-seq analysis showed a wide regulation induced by irrigation regimes on terpenes and carotenoids biosynthetic pathways. The terpene synthases and glycosyltransferases as well as genes of the network of transcription factors were upregulated, especially in berries from pre-veraison stressed vines. Since the timing and intensity of water deficit contribute to regulate berry VOCs, irrigation management can be used to achieve high-quality grapes while saving water.

Berry flavonoids are differently modulated by timing and intensities of water deficit in Vitis vinifera L. cv. Sangiovese.

In this work, we tested the effect of different regulated deficit irrigation (RDI) regimes on berry flavonoid content and its relative biosynthetic pathways. Vines were subjected to six irrigation regimes over two consecutive years: a) full irrigation during the entire irrigation period (FI); b) moderate (RDI-1M) or c) severe (RDI-1S) water deficit between berry pea-size and veraison; d) severe water deficit during the lag-phase (RDI-LS); and e) moderate (RDI-2M) or f) severe (RDI-2S) water deficit from veraison through harvest. Berries from both RDI-1 treatments showed the highest accumulation of anthocyanins, upregulating the expression of many genes of the flavonoid pathway since the beginning of veraison until harvest, far after the water deficit was released. Although to a lesser degree than RDI-1, both post-veraison water deficit treatments increased anthocyanin concentration, particularly those of the tri-substituted forms, overexpressing the F3'5'H hydroxylases. The moderate deficit irrigation treatments enhanced anthocyanin accumulation with respect to the severe ones regardless of the period when they were applied (pre- or post-veraison). The water deficit imposed during the lag-phase downregulated many genes throughout the flavonoid pathway, showing a slight reduction in anthocyanin accumulation. The measurements of cluster temperature and light exposure highlighted that under deficit irrigation conditions, the effects induced by water stress prevailed over that of light and temperature in regulating anthocyanin biosynthesis. Flavonol concentration was higher in RDI-1S berries due to the upregulation of the flavonol synthases and the flavonol-3-O-glycosyltransferases. In this case, the higher cluster light exposure induced by water deficit in RDI-1S berries had a major role in flavonol accumulation. We conclude that the timing and intensity of water stress strongly regulate the berry flavonoid accumulation and that proper management of deficit irrigation can modulate the phenylpropanoid and flavonoid pathways.

Cations and Phenolic Compounds Concentrations in Fruits of Fig Plants Exposed to Moderate Levels of Salinity.

Fig trees are often grown in areas affected by salinity problems. We investigated changes in the concentrations of 15 phenolic compounds and mineral elements (Mg, Ca, K, Zn, Cu, Mn, Mo, Fe, Na) in fruits of fig plants (Ficus carica L. cv. Dottato) subjected to irrigation with saline water (100 mM of NaCl) for 28 days. We used UHPLC-MS/MS techniques to determine chlorogenic acid, tiliroside, catechin, epicatechin (ECTC), p-coumaric acid, trans-ferulic acid, phloridzin, phloretine, quercetagetin 7-O-glucoside, rutin, quercetin 3-O-glucoside, kaempferol 3-O-rutinoside, kaempferol 7-O-glucoside, kaempferol 3-O-glucoside, and quercetin. There was a steep gradient of Na+ concentrations between the root and the canopy of salinized plants, but leaf Na+ was similar in control and salt-treated plants. Quercetin, ECTC, and chlorogenic acid were the most abundant phenolic compounds in fig fruits. Salinity increased total phenols by 5.6%, but this increase was significant only for ECTC. Salt stress significantly increased Zn and Mg concentration in the fruit. Leaf levels of K, Mg, Ca, and Mn were similar in control and salinized plants. Moderate salt stress appears to improve fig fruit quality because of its positive effect on nutrients and antioxidant compounds such as epicatechin.

Moderate Salinity Stress Affects Expression of Main Sugar Metabolism and Transport Genes and Soluble Carbohydrate Content in Ripe Fig Fruits (Ficus carica L. cv. Dottato).

Fig trees (Ficus carica L.) are commonly grown in the Mediterranean area, where salinity is an increasing problem in coastal areas. Young, fruiting plants of cv. Dottato were subjected to moderate salt stress (100 mM NaCl added to irrigation water) for 48 days before fruit sampling. To clarify the effect of salinity stress, we investigated changes in the transcription of the main sugar metabolism-related genes involved in the synthesis, accumulation and transport of soluble carbohydrates in ripe fruits by quantitative real-time PCR as well as the content of soluble sugars by quantitative 1H nuclear magnetic resonance spectroscopy. A general increase in the transcript levels of genes involved in the transport of soluble carbohydrates was observed. Alkaline-neutral and Acid Invertases transcripts, related to the synthesis of glucose and fructose, were up-regulated in ripe fruits of NaCl-stressed plants without a change in the content of D-glucose and D-fructose. The increases in sucrose and D-sorbitol contents were likely the result of the up-regulation of the transcription of Sucrose-Synthase- and Sorbitol-Dehydrogenase-encoding genes.

Corrigendum: Changes in Polyphenolic Concentrations of Table Olives (cv. Itrana) Produced Under Different Irrigation Regimes During Spontaneous or Inoculated Fermentation.

[This corrects the article DOI: 10.3389/fmicb.2018.01287.].

How an ancient, salt-tolerant fruit crop, Ficus carica L., copes with salinity: a transcriptome analysis.

Although Ficus carica L. (fig) is one of the most resistant fruit tree species to salinity, no comprehensive studies are currently available on its molecular responses to salinity. Here we report a transcriptome analysis of F. carica cv. Dottato exposed to 100 mM sodium chloride for 7 weeks, where RNA-seq analysis was performed on leaf samples at 24 and 48 days after the beginning of salinization; a genome-derived fig transcriptome was used as a reference. At day 24, 224 transcripts were significantly up-regulated and 585 were down-regulated, while at day 48, 409 genes were activated and 285 genes were repressed. Relatively small transcriptome changes were observed after 24 days of salt treatment, showing that fig plants initially tolerate salt stress. However, after an early down-regulation of some cell functions, major transcriptome changes were observed after 48 days of salinity. Seven weeks of 100 mM NaCl dramatically changed the repertoire of expressed genes, leading to activation or reactivation of many cell functions. We also identified salt-regulated genes, some of which had not been previously reported to be involved in plant salinity responses. These genes could be potential targets for the selection of favourable genotypes, through breeding or biotechnology, to improve salt tolerance in fig or other crops.

High-resolution imagery acquired from an unmanned platform to estimate biophysical and geometrical parameters of olive trees under different irrigation regimes.

The experiments were conducted in a fully-productive olive orchard (cv. Frantoio) at the experimental farm of University of Pisa at Venturina (Italy) in 2015 to assess the ability of an unmanned aerial vehicle (UAV) equipped with RGB-NIR cameras to estimate leaf area index (LAI), tree height, canopy diameter and canopy volume of olive trees that were either irrigated or rainfed. Irrigated trees received water 4-5 days a week (1348 m3 ha-1), whereas the rainfed ones received a single irrigation of 19 m3 ha-1 to relieve the extreme stress. The flight altitude was 70 m above ground level (AGL), except for one flight (50 m AGL). The Normalized Difference Vegetation Index (NDVI) was calculated by means of the map algebra technique. Canopy volume, canopy height and diameter were obtained from the digital surface model (DSM) obtained through automatic aerial triangulation, bundle block adjustment and camera calibration methods. The NDVI estimated on the day of the year (DOY) 130 was linearly correlated with both LAI and leaf chlorophyll measured on the same date (R2 = 0.78 and 0.80, respectively). The correlation between the on ground measured canopy volumes and the ones by the UAV-RGB camera techniques yielded an R2 of 0.71-0.86. The monthly canopy volume increment estimated from UAV surveys between (DOY) 130 and 244 was highly correlated with the daily water stress integral of rainfed trees (R2 = 0.99). The effect of water stress on the seasonal pattern of canopy growth was detected by these techniques in correspondence of the maximum level of stress experienced by the rainfed trees. The highest level of accuracy (RMSE = 0.16 m) in canopy height estimation was obtained when the flight altitude was 50 m AGL, yielding an R2 value of 0.87 and an almost 1:1 ratio of measured versus estimated canopy height.

Changes in Polyphenolic Concentrations of Table Olives (cv. Itrana) Produced Under Different Irrigation Regimes During Spontaneous or Inoculated Fermentation.

Irrigation is widely used for the production of table olives because it increases fruit size and yield. However, irrigation also determines less accumulation of total phenols, an increase in water content, a decrease of firmness, lower concentrations of soluble sugars in the mesocarp, thus positively or negatively affecting the fermentation process for the production of table olives. In this study we tested the hypothesis that green fruits of cultivar Itrana obtained by different irrigation regimes had different phenolic concentration that responded differentially to spontaneous or inoculated fermentation. Fruits were harvested from two orchards in the Latina province of Latium, Italy, which had been irrigated with different volumes of water during the growing season to compare the evolution of spontaneous and inoculated fermentation processes. We measured fruit characteristics at harvest, changes in the concentrations of secoiridoids and lignans, and main microbial groups abundance during fermentation. At harvest and during fermentation the concentration of phenolic compounds was higher in fruits sampled from trees that had received less water in the field. Differences were observed between spontaneous and inoculated fermentations, with a prevalence of lactic acid bacteria (LAB) in inoculated samples. In particular, oleuropein concentration completely disappeared only from samples inoculated with the two selected strains used as starters. The inoculum with selected LAB positively influenced the fermentation process of green olives, whereas the irrigation regime previously experienced by trees did not alter fermentation.

Cultivar-specific transcriptome prediction and annotation in Ficus carica L.

The availability of transcriptomic data sequence is a key step for functional genomics studies. Recently, a repertoire of predicted genes of a Japanese cultivar of fig (Ficus carica L.) was released. Because of the great phenotypic variability that can be found in this species, we decided to study another fig genotype, the Italian cv. Dottato, in order to perform comparative studies between the two cultivars and extend the pan genome of this species. We isolated, sequenced and assembled fig genomic DNA from young fruits of cv. Dottato. Then, putative gene sequences were predicted and annotated. Finally, a comparison was performed between cvs. Dottato and Horaishi predicted transcriptomes. Our data provide a resource (available at the Sequence Read Archive database under SRP109082) to be used for functional genomics of fig, in order to fill the gap of knowledge still existing in this species concerning plant development, defense and adaptation to the environment.

The Role of Polyphenoloxidase, Peroxidase, and ß-Glucosidase in Phenolics Accumulation in Olea europaea L. Fruits under Different Water Regimes.

Olive fruits and oils contain an array of compounds that contribute to their sensory and nutritional properties. Phenolic compounds in virgin oil and olive-derived products have been proven to be highly beneficial for human health, eliciting increasing attention from the food industry and consumers. Although phenolic compounds in olive fruit and oil have been extensively investigated, allowing the identification of the main classes of metabolites and their accumulation patterns, knowledge of the molecular and biochemical mechanisms regulating phenolic metabolism remains scarce. We focused on the role of polyphenoloxidase (PPO), peroxidase (PRX) and ß-glucosidase (ß-GLU) gene families and their enzyme activities in the accumulation of phenolic compounds during olive fruit development (35-146 days after full bloom), under either full irrigation (FI) or rain-fed (RF) conditions. The irrigation regime affected yield, maturation index, mesocarp oil content, fruit size, and pulp-to-pit ratio. Accumulation of fruit phenolics was higher in RF drupes than in FI ones. Members of each gene family were developmentally regulated, affected by water regime, and their transcript levels were correlated with the respective enzyme activities. During the early phase of drupe growth (35-43 days after full bloom), phenolic composition appeared to be linked to ß-GLU and PRX activities, probably through their effects on oleuropein catabolism. Interestingly, a higher ß-GLU activity was measured in immature RF drupes, as well as a higher content of the oleuropein derivate 3,4-DHPEA-EDA and verbascoside. Activity of PPO enzymes was slightly affected by the water status of trees during ripening (from 120 days after full bloom), but was not correlated with phenolics content. Overall, the main changes in phenolics content appeared soon after the supply of irrigation water and remained thereafter almost unchanged until maturity, despite fruit growth and the progressive decrease in pre-dawn leaf water potential. We suggest that enzymes involved in phenolic catabolism in the olive fruit have a differential sensitivity to soil water availability depending on fruit developmental stage.

Irrigation and fruit canopy position modify oil quality of olive trees (cv. Frantoio).

BACKGROUND: Fruit development and oil quality in Olea europaea L. are strongly influenced by both light and water availability. In the present study, the simultaneous effects of light environment and irrigation on fruit characteristics and oil quality were studied in a high-density orchard over two consecutive years. Olive fruits were harvested from three canopy positions (intercepting approximately 64%, 42% and 30% of above canopy radiation) of fully-productive trees subjected to full, deficit or complementary irrigation. RESULTS: Fruits receiving 61-67% of above canopy radiation showed the highest fruit weight, mesocarp oil content and maturation index, whereas those intercepting only 27-33% showed the lowest values. Palmitoleic and linoleic acids increased in oils obtained from fruits exposed to high light levels, whereas oleic acid and the oleic-linoleic acid ratio decreased. Neither canopy position, nor irrigation affected K232 , K270 , ΔK and the concentration of lignan in virgin olive oils (VOOs). Total phenols, 3,4-DHPEA-EDA [2-(3,4-hydroxyphenyl)ethyl (3S,4E)-4-formyl-3-(2-oxoethyl)hex-4-enoate] and p-HPEA-EDA (decarboxymethyl ligstroside-aglycone) increased in VOOs produced from fruits harvested from the top of the canopy, whereas full irrigation decreased total phenols and 3,4-DHPEA-EDA concentrations with respect to the complementary irrigation treatment. CONCLUSION: Light and water availability are crucial not only for tree productivity, but also they clearly affect olive oil quality. © 2017 Society of Chemical Industry.

VOCs-Mediated Location of Olive Fly Larvae by the Braconid Parasitoid Psyttalia concolor: A Multivariate Comparison among VOC Bouquets from Three Olive Cultivars.

Herbivorous activity induces plant indirect defenses, as the emission of herbivorous-induced plant volatiles (HIPVs), which could be used by parasitoids for host location. Psyttalia concolor is a larval pupal endoparasitoid, attacking a number of tephritid flies including B. oleae. In this research, we investigated the olfactory cues routing host location behavior of P. concolor towards B. oleae larvae infesting three different olive cultivars. VOCs from infested and healthy fruits were identified using GC-MS analyses. In two-choice behavioral assays, P. concolor females preferred infested olive cues, which also evoked ovipositional probing by female wasps. GC-MS analysis showed qualitative and quantitative differences among volatiles emitted by infested and healthy olives. Volatile emissions were peculiar for each cultivar analyzed. Two putative HIPVs were detected in infested fruits, regardless of the cultivar, the monoterpene (E)-ß-ocimene, and the sesquiterpene (E-E)-α-farnesene. Our study adds basic knowledge to the behavioral ecology of P. concolor. From an applied point of view, the field application of the above-mentioned VOCs may help to enhance effectiveness of biological control programs and parasitoid mass-rearing techniques.

Changes in olive oil volatile organic compounds induced by water status and light environment in canopies of Olea europaea L. trees.

BACKGROUND: Light and water are major factors in fruit development and quality. In this study, the effect of water and light in Olea europaea trees on volatile organic compounds (VOCs) in olive oil was studied over 2 years. Mature fruits were harvested from three zones of the canopy with different light exposure (64%, 42% and 30% of incident light) of trees subjected to full, deficit or complementary irrigation. VOCs were determined by SPME GC-MS and analysed by principal component analysis followed by discriminant analysis to partition treatment effects. RESULTS: Fruit fresh weight and mesocarp oil content decreased in zones where intercepted light was less. Low light levels significantly slowed down fruit maturation, whereas conditions of water deficit accelerated the maturation process. The presence of cyclosativene and α-muurulene was associated with water deficit, nonanal, valencene with full irrigation; α-muurulene, (E)-2-hexanal were related to low light conditions, while trans-ß-ocimene, α-copaene, (Z)-2-penten-1-ol, hexanal and nonanal to well exposed zones. The year strongly affected the VOC profile of olive oil. CONCLUSION: This is the first report on qualitative changes in VOCs induced by light environment and/or water status. This information is valuable to better understand the role of environmental factors on the sensory quality of virgin olive oil.

Oil accumulation in intact olive fruits measured by near infrared spectroscopy-acousto-optically tunable filter.

BACKGROUND: A field experiment was conducted to test the reliability of the near infrared spectroscopy (NIR)-acousto-optically tunable filter (AOTF) method to measure mesocarp oil content in vivo against nuclear magnetic resonance (NMR) determinations using three different olive cultivars at different stages of ripening. RESULTS: In the partial least squares model carried out for the cultivar 'Arbequina', the coefficient of determination in calibration (R(2)c) was 0.991, while the coefficient of determination in cross-validation (R(2)cv) was 0.979. For the cultivar 'Frantoio' the indexes were 0.982 and 0.971, respectively; while for the cultivar 'Leccino' R(2)c was 0.977 and R(2)cv was 0.965. Finally, for the combined model (sum of the three varieties) these indexes were 0.921 and 0.903, respectively. The residual predictive deviation (RPD) ratio was insufficient for the predictive model of cultivar 'Leccino' only (1.98), whereas in the other cases the RPD ratios were completely sufficient, within the estimation range over 2.5-3 (2.61 in the global model, and 4.23 in the cultivar 'Frantoio'), or in describing a large capacity with values greater than 5, as in the cultivar 'Arbequina' (9.58). CONCLUSION: NIR-AOTF spectroscopy proved to be a novel, rapid and reliable method to monitor the oil accumulation process in intact olive fruits in the field. The innovative approach of coupling NIR and NMR technologies opens up new scenarios for determining the optimal time for harvesting olive trees to obtain maximum oil production.

Water deficit-induced changes in mesocarp cellular processes and the relationship between mesocarp and endocarp during olive fruit development.

A field experiment was conducted during two consecutive growing seasons to determine and quantify the growth response of the olive (Olea europaea L. cv. Leccino) fruit and of its component tissues to tree water status. Pre-dawn leaf water potential (Psi(w)) and fruit volume were measured at about weekly intervals, and fresh weight (FW) and dry weight (DW) of the fruit tissues at 15, 20 and 21 weeks after full bloom (AFB). Fruit anatomical sections were prepared at 8, 15 and 21 weeks AFB for area determinations and cell counts. Fruit volume of the well-watered trees (average Psi(w) = -0.97 MPa) increased rapidly and reached the greatest final size, that from the most stressed (average Psi(w) = -2.81 MPa) grew most slowly and were smallest. In general, equatorial transverse areas of the mesocarp increased with increasing Psi(w), and this response was more evident at 21 than at 15 weeks AFB. By 21 weeks AFB, the mesocarp of the well-watered trees reached values more than three times higher than those measured at 8 weeks AFB. The endocarp FW and DW did not increase between 15 and 21 weeks AFB. Within each sampling date the endocarp area, FW and DW responded weakly to Psi(w). The mesocarp-to-endocarp ratio (FW and DW) increased from 15 to 21 weeks AFB regardless of water status, mainly due to the mesocarp growth. In both years at 20 and 21 weeks AFB, low values of the mesocarp-to-endocarp ratio were found with Psi(w) below -2.5 MPa. Within the mesocarp, cell size was more responsive to water deficit than to cell number. At 8 weeks AFB, the number of cells in the mesocarp was unaffected by tree water deficit, whereas cell size decreased, although slightly, in fruits sampled from trees in which Psi(w) was < -3.0 MPa. At 21 weeks AFB, cell size showed a linear decrease with increasing level of water deficit, whereas the number of cells at 21 weeks AFB decreased as the Psi(w) decreased below -2.5 MPa and seemed unaffected above that range. Overall, the results clarify the complexity of the water-induced response of mesocarp and endocarp growth and cellular processes of olive fruits.

Irrigation effects on quality, phenolic composition, and selected volatiles of virgin olive oils cv. Leccino.

Field-grown olive trees (Olea europaea L. cv. Leccino) were used over two growing seasons to determine the effect of deficit irrigation regimes on virgin olive oil (VOO) quality. Drip irrigation was managed to maintain a predawn leaf water potential (PLWP): (a) higher than -1.1 MPa (full irrigation: FI); (b) between -1.0 and -3.3 MPa (deficit irrigation: DI); (c) higher than -4.2 MPa (severe deficit irrigation: SI). The fruit yield and oil yield of DI trees were over 90% of those of FI treatments in both years, respectively, whereas yields of SI trees ranged from 61 to 76%. The irrigation regime had minor effects on the free acidity, peroxide value, and fatty acid composition of VOO. The concentrations of phenols and o-diphenols in VOO were negatively correlated with PLWP. The concentrations of the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol (3,4-DHPEA-EDA), the isomer of the oleuropein aglycon (3,4-DHPEA-EA), and the dialdehydic form of decarboxymethyl elenolic acid linked to (p-hydroxyphenyl)ethanol (p-HPEA-EDA) were lower in FI than in SI treatments. The concentrations of lignans (+)-1-acetoxipinoresinol and (+)-1-pinoresinol were unaffected by the irrigation regime. The tree water status had a marked effect on the concentration of volatile compounds, such as the C(6)-saturated and unsaturated aldehydes, alcohols, and esters.