Developmental expression of β-glucosidase in olive leaves

Plant β-glucosidases catalyze the hydrolysis of glycosidic linkages and play a vital role in defense against pathogens and stress. The present work investigated the relationship between leaf development and β-glucosidase protein content in Olea europea L. (cv. Picual) leaves. The total chlorophyll content increased with leaf age in current-season leaves. Immunoblot analysis revealed that the content of 61 kD protein of β-glucosidase also increased with leaf age, and that the enzyme existed in three isoforms (pI 5.8–6.2). Statistical analysis indicated a strong correlation between chlorophyll and β-glucosidase protein contents.     

Enhancement of enzymatic hydrolysis rate of olive oil in water by dimethyl β-cyclodextrin

Summary The hydrolysis rate of olive oil byCandida cylindracea lipase in an aqueous solution without surfactants can be increased up to 6-fold by the addition of up to 60 mg/ml of dimethyl -cyclodextrin (DMCD).     

On the origins and spread of Olea europaea L. (olive) domestication: evidence for shape variation of olive stones at Ugarit,Late Bronze Age,Syria—a window on the Mediterranean Basin and on the westward diffusion of olive varieties

Charred archaeological stones of Olea europaea L. (olive) from Late Bronze Age Ugarit, Syria, were analyzed with geometric morphometry and compared with a morphological differentiation model established on the basis of analyses of modern spontaneous (uncultivated) olive populations and cultivated varieties of various origins within the Mediterranean Basin. The results allow a reinterpretation of the east–west morphological diversity previously observed in wild olives. The archaeobotanical data were compared in detail to the partly geographically structured modern morphological diversity of the cultivated olive. Ancient morphotypes could be distinguished, among which one is dominant in the assemblage. Their diffusion from east to west is shown, and their time of arrival in the northwestern Mediterranean can be evaluated by comparison to archaeological material from that area. Combining morphometric and genetic data, modern reference and archaeological material also guides us in understanding the mechanisms that prevailed in the long-term agrobiodiversity of the olive.     

An alkaline β-glucosidase isolated from an olive brine strain of Wickerhamomyces anomalus

An efficient β-glucosidase (βG)-producing strain, Wickerhamomyces anomalus BS81, was isolated from naturally fermented olive brine and identified based on PCR/restriction fragment length polymorphism of the rDNA internal transcribed spacer and sequence analysis of the D1/D2 region of the 26S rRNA gene. The hydrolytic activity of the βG had an optimum pH of 8.5 and an optimum temperature of 35 °C. The enzyme had high substrate specificity and high catalytic efficiency (K(m) 0.99 mM, V(max) 14 U g(-1) of cells) for p-nitrophenyl-β-d-glucopyranoside. The enzyme was activated by increasing concentrations of NaCl, with maximum activity at 150 g L(-1) NaCl. Although βGs have been purified and characterized from several other sources, the W. anomalusβG is unique among βGs because its relative maximum activity occurs at alkaline pH and 35 °C. Moreover, the yeast strain has esterase activity that acts synergistically with βG to degrade oleuropein to debitter table olives and olive oil.     

Effects of olive leaf polyphenols against H₂O₂ toxicity in insulin secreting β-cells

In pancreatic β-cells, although H?O? is a metabolic signal for glucose stimulated insulin secretion, it may induce injury in the presence of increased oxidative stress (OS) as in the case of diabetic chronic hyperglycemia. Olea europea L. (olive) leaves contain polyphenolic compounds that may protect insulin-secreting cells against OS. The major polyphenolic compound in ethanolic olive leaf extract (OLE) is oleuropein (about 20%), thus we compared the effects of OLE with the effects of standard oleuropein on INS-1 cells. The cells were incubated with increasing concentrations of OLE or oleuropein for 24 h followed by exposure to H?O? (0.035 mM) for 45 min. H?O? alone resulted in a significantly decreased viability (MTT assay), depressed glucose-stimulated insulin secretion, increased apoptotic and necrotic cell death (AO/EB staining), inhibited glutathione peroxidase activity (GPx) and stimulated catalase activity that were associated with increased intracellular generation of reactive oxygen species (ROS) (fluorescence DCF). OLE and oleuropein partly improved the viability, attenuated necrotic and apoptotic death, inhibited the ROS generation and improved insulin secretion in H?O?-exposed cells. The effects of oleuropein on insulin secretion were more pronounced than those of OLE, while OLE exerted a stronger anti-cytotoxic effect than oleuropein. Unlike OLE, oleuropein had no significant preserving effect on GPx; however, both compounds stimulated the activity of catalase in H?O?-exposed cells. These findings indicate different modulatory roles of polyphenolic constituents of olive leaves on redox homeostasis that may have a role in the maintenance of β-cell physiology against OS.     

Development of 81 new polymorphic EST-derived microsatellite markers for the olive flounder,Paralichthys olivaceus

Expressed sequence tags (ESTs) can be used to identify microsatellite markers. We developed 81 polymorphic microsatellite markers from 4,940 ESTs of the olive flounder, Paralichthys olivaceus. Out of 100 EST-derived microsatellites for which PCR primers were designed, 81 loci were polymorphic in 30 individuals from a single natural population with 2–28 (mean 10.6) alleles per locus. The observed and expected heterozygosities of these loci were 0.033–1.000 and 0.033–0.965, respectively. Segregation analysis within a mapping family revealed non-amplifying null alleles at five loci. These new EST-derived microsatellite markers should be useful for population genetic analyses, pedigree tracing and constructing a linkage map for olive flounder.     

In vitro plantlet regeneration of “dwarf” Indian olive (Elaeocarpus robustus Roxb.): a fruit plant of Bangladesh

A plantlet regeneration protocol was developed on pot-grown mature plants of Elaeocarpus robustus Roxb. cv. Dwarf from nodal and leaf explants. The best yield of adventitious shoots was achieved from the leaf-derived calli in a modified MS (MMS₁, half strength of major salts, full strength of minor salts, and vitamins) medium containing 4.0 μM BA + 4.0 μM Kn + 0.5 μM NAA + 15% coconut water (CW). The shoot multiplication rate was amplified about twofold per culture after the addition of 15% CW to the medium. The rate of shoot multiplication reached maximum at the 5th subculture, and it maintained this rate throughout the 3 subsequent subcultures. The best rooting in vitro was investigated by subculturing the microcuttings in an MMS₂ (half strength of both major salts and minor salts and full strength of vitamins) medium containing 1.0 μM IBA in the dark for one initial week at 30°C, followed by subculturing them in a plant-growth regulator (PGR)-free medium in the light. The plantlets raised in vitro were successfully established under ex vitro conditions.     

Synoptic and meteorological characterisation of olive pollen transport in Córdoba province (south-western Spain)

The main goal of the present study was to provide a detailed analysis of olive pollen transport dynamics in the province of Córdoba (south-western Spain) by applying back-trajectory analysis. Pollen data from 2006 and 2007 were analysed at four monitoring sites: Córdoba city in the centre of the province, Baena and Priego de Córdoba located in the south, and El Cabril reserve (Hornachuelos Natural Park) in the north. Particular attention was paid to nine episodes of high pollen counts. Synoptic surface maps were used, and kinematic back-trajectories (3D) were computed using the hybrid single particle Lagrangian integrated trajectory model (HYSPLIT) at 500 m above ground level, run with a time-step of 6 h over a period of 36 h. Findings were analysed in conjunction with daily and bi-hourly airborne pollen data, field phenological data and hourly surface meteorological data recorded at nearby stations: temperature, relative humidity, rainfall, wind direction and wind speed. The results identified two pollen source areas over the Córdoba province, the largest one located in the south, affecting Baena, Priego de Córdoba and Córdoba city, and one smaller located in the west, which determines mainly the pollen cycle over the north of the province, El Cabril. In addition, two air mass circulations were found, one coming from the south and crossing the main olive pollen sources very close to the surface and being frequently associated with higher pollen counts, and the other coming from the west and, in the episodes investigated, influencing mainly the north of the province.     

Effects of the planting density on water relations and production of ‘Chemlali’ olive trees (Olea europaea L.)

Water relations are a key factor limiting olive production. In this study, effects of plating density on physiological aspects and productivity of ‘Chemlali’ olive trees were analyzed under rain-fed conditions in four planting densities (156, 100, 69 and 51 trees ha⁻¹), in an experimental olive orchard located in the center of Tunisia. Seasonal changes in leaf relative water content (RWC), leaf water potential, stomatal conductance (g _s), CO₂ assimilation rate and tree production were studied. Accompanying the changes in leaf water status, all the monitored trees reduced leaf stomatal conductance (g _s) and photosynthetic rate (A) throughout the summer drought, mirroring the increase in soil moisture deficit and vapor pressure deficit. However, the decrease in gas exchange was much more pronounced in high planting densities than in low ones. Our results confirm that the increase of tree-to-tree water competition with planting density was significant in the dry climate of Tunisia. Thus, planting density is critical when planting new olive orchards in arid regions.     

Are indigenous strains of Trichogramma sp. (Hym., Trichogrammatidae) better candidates for biological control of lepidopterous pests of the olive tree?

Strains of the egg parasitoid, Trichogramma, indigenous in olive groves in the Mediterranean region (T. bourarachae Pintureau & Babault, T. cacoeciae Marchal, T. cordubensis Vargas & Cabello, T. euproctidis Girault, T. nerudai Pintureau & Gerding, T. oleae Voegelé & Pointel) as well as commercially available strains (T. brassicae Bezdenko, T. cacoeciae, T. evanescens Westwood), were assessed in laboratory and semi-field experiments for their attributes as biological control agents for use against lepidopterous olive pests like the olive moth (Prays oleae Bern.) and the jasmine moth (Palpita unionalis Hübn.). In a choice test, an Egyptian strain of T. cordubensis parasitized significantly more olive and jasmine moth eggs than those of the rearing host Sitotroga cerealella Olivier. This strain was also most efficient in finding eggs of the target pests on olive foliage and on potted olive trees. Survival and fecundity of a Tunisian strain of T. bourarachae was not affected when exposed to hot (35°C) and arid (< 40% relative humidity) conditions in comparison to the mild standard of 25°C and 70% relative humidity. It was concluded that indigenous strains collected from olive groves were more effective against the target pests and more tolerant to arid conditions than commercially available parasitoid species, indicating that biological control agents should preferably be isolated from the relevant pest/crop system.     

Do extraction procedures affect olive oil quality and stability?

Quality and stability of olive oil extracted using discontinuous (pressing) or continuous (centrifuging) procedures were studied. Virgin olive oils extracted from two olive varieties were stored for 17 months. Analytical parameters of oil quality (acidity, peroxide value, polyphenol content, chlorophyll amount, fatty acid composition and triacylglycerol molecular species) were followed at different time intervals. Our results showed that no differences exist between the two types of tested mill, since the studied parameters were unchanged or varied slightly in the same range during the time of oil conservation. Changes depend on olive varieties rather than extraction systems used. Moreover, oils produced by the continuous system contain comparatively higher levels of polyphenols, which may confer to them greater resistance against oxidation for long-term storage.     

Allelic variation, balancing selection and positive selected sites detected from MHC class Iα gene of olive flounder

Major histocompatibility complex (MHC) genes play an important role in the immune response of vertebrates. Allelic polymorphism and pattern of evolution in MHC genes has been investigated in many mammals, however, much less is known in teleost. In the present study, we have investigated complete MHC Iα gene consists of 7 exons and 6 introns in Olive flounder (Paralichthys olivaceus). Genetic variation in the MHC class Iα gene was also tested in flounder. In 32 individuals, a total of 62 alleles were detected from exon 2 of MHC class Iα gene. The rate of non-synonymous substitutions (d ( N )) occurred at a significantly higher frequency than that of synonymous substitutions (d ( S )) in PBR and non-PBR, suggesting that balancing selection for maintaining polymorphisms at the MHC Iα locus. Many positive selection sites were found to act very intensively on antigen binding sites. Our founding suggests a snapshot in an evolutionary process of MHC Iα gene evolution of the P. olivaceus.     

Photosynthesis parameters and activities of enzymes of oxidative stress in two young ‘Chemlali’ and ‘Chetoui’ olive trees under water deficit

The effects of water deficit on photochemical parameters and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were investigated in two olive cultivars differing in drought tolerance — ‘Chemlali’ and ‘Chetoui’. After 30 days without irrigation, leaf water potential fell to −5.5 MPa that was accompanied by a marked decrease in net photosynthesis in ‘Chetoui’ olive cultivar. Maximal efficiency of PSII photochemistry (F_v/F_m) decreased slightly in ‘Chemlali’ (28 %) and substantially in ‘Chétoui’ (47 %). Both cultivars showed a similar decline (about 25 %) in the photochemical quenching coefficient, but only the drought-sensitive olive cultivar exhibited an enhancement (31 %) of non-photochemical fluorescence quenching under water deficit conditions. The quantum yield of electron transport decreased in both olive cultivars. ‘Chemlali’ showed a higher protection against oxidative stress, as judged from the lower levels of the malondialdehyde production. Catalase activity was higher in ‘Chetoui’. Glutathione reductase activity was increased similarly in both olive cultivars under water stress. Ascorbate peroxidase activity was enhanced in ‘Chemlali’ under water stress, but was unaffected in ‘Chetoui’. While, superoxide dismutase activity was inhibited in both cultivars under water stress, but higher activity was detected in ‘Chemlali’. Thus, the ability to increase ascorbate peroxidase and a higher superoxide dismutase activity might be an important attribute linked to the drought tolerance in ‘Chemlali’ olive cultivar.     

A meta-analysis of biocontrol potential and herbivore pressure in olive crops: Does integrated pest management make a difference?

Agricultural policies in the European Union (EU) are increasingly promoting organic management and integrated pest management (IPM) as environmentally friendly alternatives to high-input conventional management. While there is consensus that organic management is largely beneficial for biodiversity, including the natural enemies of crop pests, IPM has been much less scrutinized. We conducted a meta-analysis based on 294 observations extracted from 18 studies to compare the effects of conventional, IPM and organic management on biocontrol potential and herbivore pressure in olive, an important cash crop in the EU. Information about the management practices used was also compiled to assess differences in intensity between the three management strategies. Results suggest that IPM is predominantly based on intensive practices, employing chemical control rather than preventive measures as a first resort. Biocontrol potential and herbivore pressure were similar in conventional management and IPM. Moreover, biocontrol potential was higher in organic crops than in crops under IPM, especially when considering canopy-dwelling natural enemies. Although organic management enhanced biocontrol potential, it also benefitted some olive pests, and in both cases effects were more pronounced at warmer temperatures. Our results suggest that, in its current form, IPM might not significantly affect biocontrol potential or herbivore pressure when compared with conventional olive crop management. A shift to a more comprehensive implementation of IPM practices is thus needed, involving the use of proactive measures to promote natural enemies and regulate olive pests before resorting to chemical control. Moreover, greater use of non-chemical inputs might be required for effective regulation of olive pests in organic olive crops.     

Water-retention additives’ effects on plant water status and some physiological parameters of two olive cultivars under reduced irrigation regimes

In arid and semi-arid zones of world, the majority of annual rainfall or irrigation water is lost due to high evaporation from the soil. The amount of available water in soil can be increased by surface mulching. However, the effect varies with soils, climate and kind of mulch material used. Some physiological and biochemical parameters of two olive cultivars (‘Konservolea’ and ‘Manzanilla’) as affected by different water-retention additives (organic, chemical and mineral mulches and hydrogel) and two amounts of irrigation water were studied in pot experiments during 2013. Water stress affects plant water relations and photosynthesis and caused oxidative stress in olive. Our results confirmed that water-retention additives partly mitigated the adverse effects of water stress on olive restoring most of the measured parameters similar to those of control unstressed plants. Mulching treatment enhanced antioxidative defenses and reduced water deficit-induced oxidative damage in olive compared to zero mulching treatment. The results indicated that de-oiled olive pomace mulch and pistachio shell mulch—as new mulch materials—proved to be very effective for soil water conservation under arid conditions. It can be concluded that water management through mulch application can strongly influence the cultivar response to water deficit.     

β-Glucan synthase induction in mushrooms grown on olive mill wastewaters

-1-3-Glucan synthase activity and its induction by olive mill wastewaters (OMW) was studied in ten fungal strains (Auricularia auricula-judae, Lentinula edodes, Pleurotus eryngii, Stropharia aeruginosa, Agrocybe aegerita, P. pulmonarius, Armillaria mellea, P. ferulae, P. ostreatus, P. nebrodensis). A microtiter-based enzymatic assay on -1-3-glucan synthase activity was carried out on all mycelia growth both on the control medium and on OMW. Among the fungi assayed, L. edodes -1-3-glucan synthase was highly enhanced in OMW. The main components of OMW, i.e. phenols and lipids, were added separately to the control medium, to highlight the mechanism of L. edodes -1-3-glucan synthase induction. A Southern blot analysis and PCR with degenerated primers were carried out to detect the presence of fks1-like genes in these Basidiomycetes. The sequences obtained from the ten Basidiomycota were remarkably similar to fks1 from Filobasidiella neoformans. Spectrofluorimetric and RT-PCR analyses of -1-3-glucan synthase were performed on the mycelia of L. edodes. In this fungus, a strong stimulation of -1-3-glucan synthase mRNA and protein was recorded in the presence of OMW and phenols.     

Plant-soil interactions in a fertigated ‘Manzanilla de Sevilla’ olive orchard

Main processes governing the plant-soil interactions in adult olive (Olea europaea L.) trees under fertigation were studied to better understand the response of the trees to this agricultural practice widely used in new olive orchards. Our final objective was to obtain soundly based scientific evidences for a rational choice of the fertilizer dose. Measurements were made in a ‘Manzanilla de Sevilla’ olive orchard in which 200 g N, 400 g N and 600 g N per tree and irrigation period (T200, T400 and T600, respectively) of a 4N-1P-3K fertilizer were applied by fertigation for 5 years (1999–2003); a control treatment (unfertilized) was also established. Four years after the start of the experiment mean values of soil P and K concentrations were greater in the fertigation treatments than in the control. For K, concentrations increased with fertilizer dose. The profile of NO₃-N, P and K concentrations within the irrigation wetted zone was studied in 2003; in the top soil layer, the concentrations of the three elements increased with fertilizer dose, generally showing linear responses to the different doses; in deeper soil layers, concentrations also increased with fertigation, but to a lesser extent; the concentrations of NO₃-N, P and K recorded at 0.8–0.9 m depth in the soil of T600, together with observations of root distribution, were enough to suggest leaching losses and possible groundwater contamination. As a consequence of the higher soil nutrient availability, leaf N, P and K increased generally with dose. Leaf N deficiencies and low, but not deficient, leaf K levels were found in control trees in 2002 and 2003, as well as in T200 trees in 2003. Differences between treatments in shoot length, trunk circumference and canopy volume were not significant, for any studied year. Nevertheless, between February 1999 and November 2003 there was a significant increase in canopy volume with fertilizer dose. In 2003, fruit yield increased with fertilizer dose, as a consequence of an increase both in fruit number and weight. Cumulative yield for the experimental period also increased with fertilizer dose. These results are further evidence to confirm previous research made with the same experimental set-up: T400 for oil quality and T600 for table olive quality seem to be the most appropriate treatments, although there is a risk for leaching losses and the possibility of groundwater contamination with T600.