Effect of photostimulation through red LED light radiation on natural fermentation of table olives: An innovative case study with Negrinha the Freixo variety.

In this study, for the first time, red LED light radiation was applied to the fermentation process of table olives using the Negrinha de Freixo variety. Photostimulation using LED light emission (630 ± 10 nm) is proposed to shorten and speed up this stage and reduce time to market. Several physical-chemical characteristics and microorganisms (total microbial count of mesophilic aerobic, molds, yeasts, and lactic acid bacteria) and their sequence during fermentation were monitored. The fermentation occurred for 122 days, with two irradiation periods for red LED light. The nutritional composition and sensory analysis were performed at the end of the process. Fermentation under red LED light increased the viable yeast and lactic acid bacteria (LAB) cell counts and decreased the total phenolics in olives. Even though significant differences were observed in some color parameters, the hue values were of the same order of magnitude and similar for both samples. Furthermore, the red LED light did not play a relevant change in the texture profile, preventing the softening of the fruit pulp. Similarly, LED light did not modify the existing type of microflora but increased species abundance, resulting in desirable properties and activities. The species identified were yeasts - Candida boidinii, Pichia membranifaciens, and Saccharomyces cerevisiae, and bacteria - Lactobacillus plantarum and Leuconostoc mesenteroides, being the fermentative process dominated by S. cerevisiae and L. plantarum. At the end of fermentation (122 days), the irradiated olives showed less bitterness and acidity, higher hardness, and lower negative sensory attributes than non-irradiated. Thus, the results of this study indicate that red LED light application can be an innovative technology for table olives production.

Olive Varieties under UV-B Stress Show Distinct Responses in Terms of Antioxidant Machinery and Isoform/Activity of RubisCO.

In recent decades, atmospheric pollution led to a progressive reduction of the ozone layer with a consequent increase in UV-B radiation. Despite the high adaptation of olive trees to the Mediterranean environment, the progressive increase of UV-B radiation is a risk factor for olive tree cultivation. It is therefore necessary to understand how high levels of UV-B radiation affect olive plants and to identify olive varieties which are better adapted. In this study we analyzed two Italian olive varieties subjected to chronic UV-B stress. We focused on the effects of UV-B radiation on RubisCO, in terms of quantity, enzymatic activity and isoform composition. In addition, we also analyzed changes in the activity of antioxidant enzymes (SOD, CAT, GPox) to get a comprehensive picture of the antioxidant system. We also evaluated the effects of UV-B on the enzyme sucrose synthase. The overall damage at biochemical level was also assessed by analyzing changes in Hsp70, a protein triggered under stress conditions. The results of this work indicate that the varieties (Giarraffa and Olivastra Seggianese) differ significantly in the use of specific antioxidant defense systems, as well as in the activity and isoform composition of RubisCO. Combined with a different use of sucrose synthase, the overall picture shows that Giarraffa optimized the use of GPox and opted for a targeted choice of RubisCO isoforms, in addition to managing the content of sucrose synthase, thereby saving energy during critical stress points.

Physiological and Metabolite Reconfiguration of Olea europaea to Cope and Recover from a Heat or High UV-B Shock.

To understand how olives reconfigure their metabolism to face stress shock episodes, plants from the economically relevant olive (Olea europaea cv. Cobrançosa) were exposed to high UV-B radiation (UV-B, 12 kJ m-2 d-1) or heat shock (HS, 40 °C) for two consecutive days. The physiological responses and some important lipophilic compounds were evaluated immediately (day 0) and 30 days after UV-B or HS episodes. Both treatments induced a reduction of the olive physiological performance, particularly increasing cell membrane damages and proline pool and at the same time reducing chlorophyll levels, the quantum yield of photosystem II (ΦPSII), and the efficiency of excitation energy capture by open photosystem II (PSII) reaction centers (F'v/F'm). Nevertheless, the HS episode caused more adverse effects, additionally reducing the pool of protective pigments (carotenoids) and the maximum efficiency of PSII (with F0 increase). In the UV-B treatment, despite the higher lipid peroxidation, the activation of some stress protective mechanisms (e.g., increase of NPQ and carotenoids and remobilization of some metabolites, such as phytol and proline) might have contributed to avoiding photoinhibition. Thirty days after stress relief, the performance of olives from both treatments recovered similarly, in part due to the metabolites' adjustments that contributed to strengthened stress protection (an increase of long-chain alkanes) and provided energy (through the use of soluble sugars, mannitol, and myo-inositol) for re-establishment. Other metabolites, like anthocyanins and squalene, also have an important role in responding specifically to HS or UV-B recovery for helping in the oxidative damage control. These data contribute to understanding how young olive plants may deal with climatic episodes when being transferred from nurseries to field orchards, under the actual context of climate change.

Feasible production of biomass and natural antioxidants through callus cultures in response to varying light intensities in olive (Olea europaea. L) cult. Arbosana.

Light is the most important physical factor in growth and development of plants. Light intensity is directly proportional to the growth and accumulation of natural antioxidants during in vitro cultures of various medicinal plants. The present research study was designed to determine the effect of different light intensities i.e. normal light (2000-2500 lx), diffused light (500-1000 lx) and complete dark (0 lx) on callus growth dynamics and production of natural antioxidants in olive cult. Arbosana. Highest callus induction frequency (50%) was observed in the stem explants pre-treated with silver nanoparticles suspension (AgNPs: 50 ppm) and cultured on MS media supplemented with combination of 6-Benzylaminopurine (BAP: 2 mg/l), Gibberellic acid (GA3: 1.5 mg/l) plus Naphthalene acetic acid (NAA: 0.5 mg/l). Maximum callus biomass (FW = 1414 mg/l) was recorded when the cultured explants were incubated initially for seven days in complete darkness, followed by transference to diffused light for one week and then finally placed under normal light in total fifty six days culture period. Moreover, phytochemical analysis of the callus cultures showed significantly higher activities of antioxidant enzymes i.e. SOD, POD, CAT and APx (2.45, 2.96, 2.57 and 1.67 U/mg. protein) in the callus cultures grown under dark condition as compared with other light treatments. For non-enzymatic antioxidant potential, maximum activity of TPC, TFC, PAL and DPPH (2.42 mg GAE/g, 1.50 mg QAE/g, 3.95 U/mg and 75%) were recorded in the calli raised in vitro under diffused light. This is the first report on the production of natural antioxidants in response to different light intensities in callus cultures of Olea europaea. Future studies should focus on large scale production of callus cultures in order to yield maximum biomass from this high valued plant.

UV-B radiation modulates physiology and lipophilic metabolite profile in Olea europaea.

Ultraviolet-B (UV-B) radiation plays an important role in plant photomorphogenesis. Whilst the morpho-functional disorders induced by excessive UV irradiation are well-known, it remains unclear how this irradiation modulates the metabolome, and which metabolic shifts improve plants' tolerance to UV-B. In this study, we use an important Mediterranean crop, Olea europaea, to decipher the impacts of enhanced UV-B radiation on the physiological performance and lipophilic metabolite profile. Young olive plants (cv. 'Galega Vulgar') were exposed for five days to UV-B biologically effective doses of 6.5 kJ m-2 d-1 and 12.4 kJ m-2 d-1. Cell cycle/ploidy, photosynthesis and oxidative stress, as well as GC-MS metabolites were assessed. Both UV-B treatments impaired net CO2 assimilation rate, transpiration rate, photosynthetic pigments, and RuBisCO activity, but 12.4 kJ m-2 d-1 also decreased the photochemical quenching (qP) and the effective efficiency of PSII (ΦPSII). UV-B treatments promoted mono/triperpene pathways, while only 12.4 kJ m-2 d-1 increased fatty acids and alkanes, and decreased geranylgeranyl-diphosphate. The interplay between physiology and metabolomics suggests some innate ability of these plants to tolerate moderate UV-B doses (6.5 kJ m-2 d-1). Also their tolerance to higher doses (12.4 kJ m-2 d-1) relies on plants' metabolic adjustments, where the accumulation of specific compounds such as long-chain alkanes, palmitic acid, oleic acid and particularly oleamide (which is described for the first time in olive leaves) play an important protective role. This is the first study demonstrating photosynthetic changes and lipophilic metabolite adjustments in olive leaves under moderate and high UV-B doses.

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.

The expression of light-related leaf functional traits depends on the location of individual leaves within the crown of isolated Olea europaea trees.

BACKGROUND: The spatial arrangement and expression of foliar syndromes within tree crowns can reflect the coupling between crown form and function in a given environment. Isolated trees subjected to high irradiance and concomitant stress may adjust leaf phenotypes to cope with environmental gradients that are heterogeneous in space and time within the tree crown. The distinct expression of leaf phenotypes among crown positions could lead to complementary patterns in light interception at the crown scale. METHODS: We quantified eight light-related leaf traits across 12 crown positions of ten isolated Olea europaea trees in the field. Specifically, we investigated whether the phenotypic expression of foliar traits differed among crown sectors and layers and five periods of the day from sunrise to sunset. We investigated the consequences in terms of the exposed area of the leaves at the tree scale during a single day. KEY RESULTS: All traits differed among crown positions except the length-to-width ratio of the leaves. We found a strong complementarity in the patterns of the potential exposed area of the leaves among day periods as a result of a non-random distribution of leaf angles across the crown. Leaf exposure at the outer layer was below 60 % of the displayed surface, reaching maximum interception during morning periods. Daily interception increased towards the inner layer, achieving consecutive maximization from east to west positions within the crown, matching the sun's trajectory. CONCLUSIONS: The expression of leaf traits within isolated trees of O. europaea varies continuously through the crown in a gradient of leaf morphotypes and leaf angles depending on the exposure and location of individual leaves. The distribution of light-related traits within the crown and the complementarity in the potential exposure patterns of the leaves during the day challenges the assumption of low trait variability within individuals.

Canopy light heterogeneity drives leaf anatomical, eco-physiological, and photosynthetic changes in olive trees grown in a high-density plantation.

In the field, leaves may face very different light intensities within the tree canopy. Leaves usually respond with light-induced morphological and photosynthetic changes, in a phenomenon known as phenotypic plasticity. Canopy light distribution, leaf anatomy, gas exchange, chlorophyll fluorescence, and pigment composition were investigated in an olive (Olea europaea, cvs. Arbequina and Arbosana) orchard planted with a high-density system (1,250 trees ha(-1)). Sampling was made from three canopy zones: a lower canopy (<1 m), a central one (1-2 m), and an upper one (>2 m). Light interception decreased significantly in the lower canopy when compared to the central and top ones. Leaf angle increased and photosynthetic rates and non-photochemical quenching (NPQ) decreased significantly and progressively from the upper canopy to the central and the lower canopies. The largest leaf areas were found in the lower canopy, especially in the cultivar Arbequina. The palisade and spongy parenchyma were reduced in thickness in the lower canopy when compared to the upper one, in the former due to a decrease in the number of cell layers from three to two (clearly distinguishable in the light and fluorescence microscopy images). In both cultivars, the concentration of violaxanthin-cycle pigments and ß-carotene was higher in the upper than in the lower canopy. Furthermore, the de-epoxidized forms zeaxanthin and antheraxanthin increased significantly in those leaves from the upper canopy, in parallel to the NPQ increases. In conclusion, olive leaves react with morphological and photosynthetic changes to within-crown light gradients. These results strengthen the idea of olive trees as "modular organisms" that adjust the modules morphology and physiology in response to light intensity.

Coping with drought-induced xylem cavitation: coordination of embolism repair and ionic effects in three Mediterranean evergreens.

Embolism repair and ionic effects on xylem hydraulic conductance have been documented in different tree species. However, the diurnal and seasonal patterns of both phenomena and their actual role in plants' responses to drought-induced xylem cavitation have not been thoroughly investigated. This study provides experimental evidence of the ability of three Mediterranean species to maintain hydraulic function under drought stress by coordinating the refilling of xylem conduits and ion-mediated enhancement of stem hydraulic conductance (K stem). Vessel grouping indices and starch content in vessel-associated parenchyma cells were quantified to verify eventual correlations with ionic effects and refilling, respectively. Experiments were performed on stems of Ceratonia siliqua L., Olea europaea L. and Laurus nobilis L. Seasonal, ion-mediated changes in K stem (ΔK stem) and diurnal and/or seasonal embolism repair were recorded for all three species, although with different temporal patterns. Field measurements of leaf specific stem hydraulic conductivity showed that it remained quite constant during the year, despite changes in the levels of embolism. Starch content in vessel-associated parenchyma cells changed on diurnal and seasonal scales in L. nobilis and O. europaea but not in C. siliqua. Values of ΔK stem were significantly correlated with vessel multiple fraction values (the ratio of grouped vessels to total number of vessels). Our data suggest that the regulation of xylem water transport in Mediterranean plants relies on a close integration between xylem refilling and ionic effects. These functional traits apparently play important roles in plants' responses to drought-induced xylem cavitation.

Degradations of model compounds representing some phenolics in olive mill wastewater via electro-Fenton and photoelectro-Fenton treatments.

The electrochemical oxidation of vanillic acid, o-coumaric acid and protocatechuic acid, three representative toxic phenolics in olive mill wastewater, was studied using carbon felt cathode in the electro-Fenton system. Results obtained, in the presence or absence of UV support, were compared throughout the degradation processes up to mineralization. It was demonstrated that all three phenolic compounds reacted completely with hydroxyl radicals and degraded efficiently. It was shown in the photoelectro-Fenton process that the degradation and mineralization efficiency of the phenolic compounds were enhanced by the effect of UV light, especially at the later stages of the degradation processes.

Temporal matching among diurnal photosynthetic patterns within the crown of the evergreen sclerophyll Olea europaea L.

Trees are modular organisms that adjust their within-crown morphology and physiology in response to within-crown light gradients. However, whether within-plant variation represents a strategy for optimizing light absorption has not been formally tested. We investigated the arrangement of the photosynthetic surface throughout one day and its effects on the photosynthetic process, at the most exposed and most sheltered crown layers of a wild olive tree (Olea europaea L.). Similar measurements were made for cuttings taken from this individual and grown in a greenhouse at contrasted irradiance-levels (100 and 20% full sunlight). Diurnal variations in light interception, carbon fixation and carbohydrate accumulation in sun leaves were negatively correlated with those in shade leaves under field conditions when light intensity was not limiting. Despite genetic identity, these complementary patterns were not found in plants grown in the greenhouse. The temporal disparity among crown positions derived from specialization of the photosynthetic behaviour at different functional and spatial scales: architectural structure (crown level) and carbon budget (leaf level). Our results suggest that the profitability of producing a new module may not only respond to construction costs or light availability, but also rely on its spatio-temporal integration within the productive processes at the whole-crown level.

Antioxidant defences and oxidative damage in salt-treated olive plants under contrasting sunlight irradiance.

The interactive effects of root-zone salinity and sunlight on leaf biochemistry, with special emphasis on antioxidant defences, were analysed in Olea europaea L. cv. Allora, during the summer period. Plants were grown outside under 15% (shade plants) or 100% sunlight (sun plants) and supplied with 0 or 125 mM NaCl. The following measurements were performed: (1) the contribution of ions and soluble carbohydrates to osmotic potentials; (2) the photosystem II (PSII) photochemistry and the photosynthetic pigment concentration; (3) the concentration and the tissue-specific distribution of leaf flavonoids; (4) the activity of antioxidant enzymes; and (5) the leaf oxidative damage. The concentrations of Na(+) and Cl(-) were significantly greater in sun than in shade leaves, as also observed for the concentration of the 'antioxidant' sugar-alcohol mannitol. The de-epoxidation state of violaxanthin-cycle pigments increased in response to salinity stress in sun leaves. This finding agrees with a greater maximal PSII photochemistry (F(v)/F(m)) at midday, detected in salt-treated than in control plants, growing in full sunshine. By contrast, salt-treated plants in the shade suffered from midday depression in F(v)/F(m) to a greater degree than that observed in control plants. The high concentration of violaxanthin-cycle pigments in sun leaves suggests that zeaxanthin may protect the chloroplast from photo-oxidative damage, rather than dissipating excess excitation energy via non-photochemical quenching mechanisms. Dihydroxy B-ring-substituted flavonoid glycosides accumulate greatly in the mesophyll, not only in the epidermal cells, in response to high sunlight. The activity of antioxidant enzymes varied little because of sunlight irradiance, but declined sharply in response to high salinity in shade leaves. Interestingly, control and particularly salt-treated plants in the shade underwent greater oxidative damage than their sunny counterparts. These findings, which conform to the evolution of O. europaea in sunny environments, suggest that under partial shading, the antioxidant defence system may be ineffective to counter salt-induced oxidative damage.

Seasonal evolution of diffusional limitations and photosynthetic capacity in olive under drought.

This study tests the hypothesis that diffusional limitation of photosynthesis, rather than light, determines the distribution of photosynthetic capacity in olive leaves under drought conditions. The crowns of four olive trees growing in an orchard were divided into two sectors: one sector absorbed most of the radiation early in the morning (MS) while the other absorbed most in the afternoon (AS). When the peak of radiation absorption was higher in MS, air vapour pressure deficit (VPD) was not high enough to provoke stomatal closure. In contrast, peak radiation absorption in AS coincided with the daily peak in VPD. In addition, two soil water treatments were evaluated: irrigated trees (I) and non-irrigated trees (nI). The seasonal evolution of leaf water potential, leaf gas exchange and photosynthetic capacity were measured throughout the tree crowns in spring and summer. Results showed that stomatal conductance was reduced in nI trees in summer as a consequence of soil water stress, which limited their net assimilation rate. Olive leaves displayed isohydric behaviour and no important differences in the diurnal course of leaf water potentials among treatments and sectors were found. Seasonal diffusional limitation of photosynthesis was mainly increased in nI trees, especially as a result of stomatal limitation, although mesophyll conductance (g(m)) was found to decrease in summer in both treatments and sectors. A positive relationship between leaf nitrogen content with both leaf photosynthetic capacity and the daily integrated quantum flux density was found in spring, but not in summer. The relationship between photosynthetic capacity and g(m) was curvilinear. Leaf temperature also affected to g(m) with an optimum temperature at 29 degrees C. AS showed larger biochemical limitation than MS in August in both treatments. All these suggest that both diffusional limitation and the effect of leaf temperature could be involved in the seasonal reduction of photosynthetic capacity of olive leaves. This work highlights the need for models of plant growth and ecosystem function to incorporate new parameters affecting the distribution of photosynthetic capacity in canopies.

High-power ultrasound in olive paste pretreatment. Effect on process yield and virgin olive oil characteristics.

The effect of high-power ultrasound on olive paste, on laboratory thermo-mixing operations for virgin olive oil extraction, has been studied. Direct sonication by an ultrasound probe horn (105 W cm(-2) and 24 kHz) and indirect sonication with an ultrasound-cleaning bath (150 W and 25 kHz) were applied and their effects compared with the conventional thermal treatment. A quick-heating of olive paste, from ambient (12-20 degrees C) to optimal temperature conditions (28-30 degrees C), and an oil extractability improvement were observed when applying sonication. Better extractability was obtained by direct sonication for high moisture olives (>50%) whereas indirect sonication gave greater extractability for low moisture olive fruits (<50%). Optimal application of ultrasound was achieved with direct sonication for 4 min at the beginning of paste malaxation and with indirect sonication during the malaxation time. Effect of high-power ultrasound on oil quality parameters and nutritional and sensory characteristics were studied. Changes in quality parameters (free acidity value, peroxide value, K270 and K232) were not found, however significant effects on the levels of bitterness, polyphenols, tocopherols (vitamin E), chlorophyll and carotenoids were observed. Oils from sonicated pastes showed lower bitterness and higher content of tocopherols, chlorophylls and carotenoids. Related to sensory characteristics, off-flavour volatiles were not detected in oils from sonication treatments. Total peak areas of volatiles and the ratio hexanal/E-2-hexenal, as determined by SPME analysis, were lower than non-sonicated reference oils; sensory evaluation by panel test showed higher intensity of positive attributes and lesser of negative characteristics than those untreated.