Description of the volatile fraction of Erica honey from the northwest of the Iberian Peninsula.

Heather honey is highly appreciated by consumers for its sensorial profile, which varies depending on the flora used by the honeybees. Volatile compounds contribute to these qualities. Characterisation of the volatile profile related to the botanical origin is of great interest for the standardization of unifloral honey. For this reason, 33 heather honey samples from northwest of the Iberian Peninsula were analysed by headspace solid-phase microextraction (HS-SPME) to identify the key volatile compounds in this type of honey. The aim of this research was to provide a descriptive analysis of these compounds, and to find whether there is any relationship with the main Erica species. A total of 58 volatile organic compounds were found, with hotrienol, phenylacetaldehyde, and cis-linalool being the most abundant. A principal component analysis and Spearman's rank correlation showed the homogeneity of the volatile profile in the samples, and their close relationship with the main pollen types.

Phenolic Compounds in Poorly Represented Mediterranean Plants in Istria: Health Impacts and Food Authentication.

Phenolic compounds are well-known bioactive compounds in plants that can have a protective role against cancers, cardiovascular diseases and many other diseases. To promote local food development, a comprehensive overview of the phenolic compounds' composition and their impact on human health from typical Mediterranean plants such as Punica granatum L., Ziziphus jujuba Mill., Arbutus unedo L., Celtis australis L., Ficus carica L., Cynara cardunculus var. Scolymus L. is provided. Moreover, the potential use of these data for authenticity determination is discussed. Some of the plants' phenolic compounds and their impact to human health are very well determined, while for others, the data are scarce. However, in all cases, more data should be available about the content, profile and health impacts due to a high variation of phenolic compounds depending on genetic and environmental factors. Quantifying variation in phenolic compounds in plants relative to genetic and environmental factors could be a useful tool in food authentication control. More comprehensive studies should be conducted to better understand the importance of phenolic compounds on human health and their variation in certain plants.

Chemical Composition and Biological Activities of Lebanese Pentapera Plant.

BACKGROUND Due to their chemical constituents and biological properties, plants have long been used to control life-threatening diseases. The flora of Lebanon includes many plants that have already been demonstrated to have medicinal value, and other species, such as Pentapera sicula libanotica, that are yet to be characterized. The present study characterized the chemical composition, anti-oxidant, anti-inflammatory, and anti-proliferative potential of aqueous, ethanol, and methanol extracts derived from the leaves of the Lebanese Pentapera plant. MATERIAL AND METHODS High-performance liquid chromatography (HPLC) was used to determine the chemical composition. Gas chromatography (GC) coupled with mass spectrometry (MS) was applied to determine the content of essential oil. DPPH radical scavenging assay was performed to evaluate the anti-oxidant potential. The anti-inflammatory potential was assessed using quantitative real-time PCR (qRT-PCR) by measuring TNF-alpha, IL-6, and CCL4 mRNA levels, and we assessed Cox-2 and iNOS proteins levels using Western blot (WB) analysis. MTT assay was carried out to determine the anti-proliferative potential. RESULTS We identified, mainly in the alcoholic (methanol and ethanol) extracts, distinct bioactive compounds with pharmacological relevance. In parallel, with their phytochemical content, these 2 extracts showed significant anti-oxidant, anti-inflammatory and anti-proliferative capacities. CONCLUSIONS Pentapera sicula libanotica appears to be a promising pharmacological tool.

Leaf biochemical adjustments in two Mediterranean resprouter species facing enhanced UV levels and reduced water availability before and after aerial biomass removal.

Effects of supplemented UV radiation and diminished water supply on the leaf concentrations of phenols and antioxidants of two Mediterranean resprouter species, Arbutus unedo and Quercus suber, were assessed before and after entire aerial biomass removal. Potted seedlings of both species were grown outdoors for 8 months with enhanced UV-A + UV-B, enhanced UV-A or ambient UV, in combination with two watering conditions (field capacity or watering reduction). After this period, all aerial biomass was removed and new shoots (resprouts) developed for a further 8 months under the two treatments. In general, the investment in leaf phenols was substantially greater in A. unedo than in Q. suber, while Q. suber allocated more resources to non-phenolic antioxidants (ascorbate and glutathione). In response to enhanced UV-B radiation, Q. suber leaves rose their UV-screening capacity mainly via accumulation of kaempferols, accompanied by an increased concentration of rutins, being these effects exacerbated under low-watering conditions. Conversely, A. unedo leaves responded to UV-B radiation reinforcing the antioxidant machinery by increasing the overall amount of flavonols (especially quercetins) in seedlings, and of ascorbate and glutathione, along with catalase activity, in resprouts. Nevertheless, UV effects on the amount/activity of non-phenolic antioxidants of A. unedo resprouts were modulated by water supply. Indeed, the highest concentration of glutathione was found under the combination of enhanced UV-B radiation and reduced watering, suggesting an enlargement of the antioxidant response in A. unedo resprouts. Different biochemical responses to enhanced UV and drier conditions in seedlings and resprouts of these two species might modulate their competitive interactions in the near future.

Active proton efflux, nutrient retention and boron-bridging of pectin are related to greater tolerance of proton toxicity in the roots of two Erica species.

BACKGROUND AND AIMS: Tolerance to soil acidity was studied in two species of Ericaceae that grow in mine-contaminated soils (S Portugal, SW Spain) to find out if there are interspecific variations in H+ tolerance which might be related to their particular location. METHODS: Tolerance to H+ toxicity was tested in nutrient solutions using seeds collected in SW Spain. Plant growth and nutrient contents in leaves, stems and roots were determined. Viability tests and proton exchange were studied in roots exposed, short-term, to acidic conditions. Membrane ATPase activity and the cell-wall pectic polysaccharide domain rhamnogalacturonan-II (RG-II) were analysed to find out interspecific differences. RESULTS: Variation in survival, growth and mineral composition was found between species. The H+-tolerant species (Erica andevalensis) showed greater concentration of nutrients than E. australis. Very low pH (pH 2) produced a significant loss of root nutrients (K, P, Mg) in the sensitive species. Root ATPase activity was slightly higher in the tolerant species with a correspondingly greater H+ efflux capacity. In both species, the great majority of the RG-II domains were in their boron-bridged dimeric form. However, shifting to a medium of pH 2 caused some of the boron bridges to break in the sensitive species. CONCLUSIONS: Variation in elements linked to the cell wall-membrane complex and the stability of their components (RG-II, H+-ATPases) are crucial for acid stress tolerance. Thus, by maintaining root cell structure, active proton efflux avoided toxic H+ build-up in the cytoplasm and supported greater nutrient acquisition in H+-tolerant species.

EFFECTS OF COLD STRATIFICATION AND GA3 ON GERMINATION OF ARBUTUS UNEDO SEEDS OF THREE PROVENANCES.

BACKGROUND: Arbutus unedo is a valuable Mediterranean shrub as an ornamental plant as well as fruit tree. Fresh fruits of A. unedo are a good source of antioxidants, of vitamins C, E and carotenoids and also are characterized by the high content of mineral elements. MATERIALS AND METHODS: The effects of gibberellic acid (GA3) and cold stratification (CS) on seed germination performance were investigated in A. unedo seeds collected from three provenances in the Northern part of Greece. Seeds of each provenance were soaked in solutions of GA3 (500, 1000 or 2000 ppm) for 24 h and subsequently were subjected to CS at 3 - 5°C for 0, 1, 2, and 3 months. RESULTS: Non-stratified seeds of the three A. unedo provenances which were not treated with GA3 solutions exhibited very low germination. However, seed germination was significantly improved after a one-month period of CS. Similarly, the non-stratified seeds of all three provenances became non-dormant after the treatment with 2000 ppm GA3 and they germinated at high percentages. However, in untreated seeds with GA3, after a one-month CS period the seeds of the Pieria provenance exhibited higher germination percentage than that of Rodopi provenance seeds. Furthermore, in non-stratified seeds, the Pieria provenance seeds treated with GA3 germinated at higher percentages and more rapidly than those of the other two provenances. CONCLUSION: The results indicated that untreated seeds exhibited very low germination at 20/25°C. However, in all three provenances seed germinability was significantly improved by a one-month period of CS or treatment of seeds with 2000 ppm GA3. Furthermore, there was a considerable variability among seed provenances in response to the treatments which were applied.

Understanding of human metabolic pathways of different sub-classes of phenols from Arbutus unedo fruit after an acute intake.

Arbutus unedo is a small Mediterranean fruit, commonly named strawberry tree, which is a rich source of different sub-classes of phenolic compounds, the more representative being the gallic acid derivatives, including its mono and oligomeric forms esterified with quinic and shikimic acids. In addition, galloyl derivatives, particularly gallotannins, described in A. unedo, are part of a very selective phenolic group, present in a reduced number of plant-products. The aim of the present study is to provide a better understanding of human metabolic pathways of different sub-classes of phenols from the A. unedo fruit after an acute intake by healthy adults. Therefore, the A. unedo phenolic metabolites were studied in whole blood samples (0 to 24 h), urine (24 h) and feces (12 and 24 h). Special focus was placed on the application of dried blood spot (DBS) cards for the sample collection and for the analysis of phenolic metabolites in whole blood samples. The results of the blood analysis revealed two peaks for the maximum concentrations of the main phenolic metabolites. Furthermore, it is appropriate to highlight the application of DBS cards as an efficient and accurate way to collect blood samples in post-prandial bioavailability studies. The analysis of urine (24 h) gave a wide range of phenolic metabolites showing the extensive metabolism that A. unedo phenolic compounds underwent in the human body. The results of the study provide a relevant contribution to the understanding of the in vivo human bioavailability of phenolic compounds, especially galloyl derivatives, a singular phenolic sub-group present in the A. unedo fruit.

Nutrient resorption of two evergreen shrubs in response to long­term fertilization in a bog.

Plant resorption of multiple nutrients during leaf senescence has been established but stoichiometric changes among N, P and K during resorption and after fertilization are poorly understood. We anticipated that increased N supply would lead to further P limitation or co-limitation with N or K [i.e. P-(co)limitation], decrease N resorption and increase P and K resorption, while P and K addition would decrease P and K resorption and increase N resorption. Furthermore, Ca would accumulate while Mg would be resorbed during leaf senescence, irrespective of fertilization. We investigated the effect of N, P and K addition on resorption in two evergreen shrubs (Chamaedaphne calyculata and Rhododendron groenlandicum) in a long-term fertilization experiment at Mer Bleue bog, Ontario, Canada. In general, N addition caused further P-(co)limitation, increased P and K resorption efficiency but did not affect N resorption. P and K addition did not shift the system to N limitation and affect K resorption, but reduced P resorption proficiency. C. calyculata resorbed both Ca and Mg while R. groenlandicum resorbed neither. C. calyculata showed a higher resorption than R. groenlandicum, suggesting it is better adapted to nutrient deficiency than R. groenlandicum. Resorption during leaf senescence decreased N:P, N:K and K:P ratios. The limited response of N and K and the response of P resorption to fertilization reflect the stoichiometric coupling of nutrient cycling, which varies among the two shrub species; changes in species composition may affect nutrient cycling in bogs.

Comparison of arsenic resistance in Mediterranean woody shrubs used in restoration activities.

Myrtus communis, Arbutus unedo and Retama sphaerocarpa are Mediterranean shrubs widely used in revegetation of semiarid degraded soils. The aim of this work is to study the resistance of these plants to arsenic under controlled conditions, in order to evaluate their potential use in revegetation and/or phytoremediation of As-polluted soils. R. sphaerocarpa showed higher resistance to As than M. communis or A. unedo according to its higher EC50, P status and P/As molar ratio in both, roots and shoots, and the lower increases in lipid peroxidation and decrease of chlorophyll levels in response to arsenic, while the highest arsenate sensitivity was obtained for A. unedo. Arsenic was mainly retained in roots, and, although M. communis accumulated higher arsenic amounts than the other two species, R. sphaerocarpa showed the highest root to shoot transfer. Most of the studied parameters (chlorophylls, MDA and total thiols) showed significant correlation with arsenic concentration in roots and leaves of plants, so they can be useful indexes in the diagnosis of arsenic toxicity in these species. According to our results, both M. communis and R. sphaerocarpa could be used in the revegetation of moderately arsenic contaminated sites.

Influence of pH on copper and zinc sensitivity of ericoid mycobionts in vitro.

The effect of pH on growth, metal uptake and toxicity in four isolates of ericoid mycobionts (two Hymenoscyphus ericae from unpolluted heathland sites and two H. ericae-type mycobionts from metal-contaminated mine spoil) was assessed in vitro. These isolates were incubated in liquid medium (10% Rorison's medium, glucose at 10 g l(-1)) containing either 0.25 mM Cu or 2.0 mM Zn and adjusted to pH 2, 3, 4, 5 or 6. After 30 days incubation, dry mass and mycelial metal content were determined and growth was expressed as tolerance index, i.e. dry mass in the presence of metal as a percentage of dry mass in the absence of metal. Initial medium pH had a significant effect on both tolerance index and metal accumulation. Tolerance indices were highest at pH 2, with several isolates showing a stimulation of growth (i.e. tolerance index >100%) at this pH. Tolerance index decreased at higher initial pH values and growth of two mycobionts was completely inhibited (tolerance index=0) in the Cu-supplemented media at pH 6. Reduction in tolerance index coincided with an increase in mycelial accumulation of Cu and Zn. Practical and environmental implications of these results are discussed.

Nutritional influences on the solubilization of metal phosphate by ericoid mycorrhizal fungi.

Four ericoid mycobionts (two isolates of Hymenoscyphus ericae, and two dark, sterile ericoid mycobionts isolated from metal-contaminated mine sites) were grown on solid agar plates supplemented with zinc phosphate (0.25 %) containing different forms of nitrogen (nitrate, ammonium or alanine) and different concentrations of carbon (glucose) and phosphorus (K2HPO4). The influence of nutrient variation on solubilizing ability of the fungi was assessed by measuring the zones of solubilization appearing beneath the growing colonies. All four mycobionts were capable of zinc phosphate solubilization in the presence of all three nitrogen sources and in media containing no nitrogen. No solubilization was observed at 0 mM glucose-C but was observed with increasing glucose concentration from 300 to 600 mM C. Increasing phosphorus concentration (0-5 mM P) had no effect on the solubilizing ability of the isolates. All but one of the mycobionts were capable of solubilizing calcium phosphate (CaHPO4), while no solubilization was observed in media containing aluminium phosphate (AlPO4), iron phosphate (FePO4 x 4H2O) or copper phosphate (Cu3O8P2 x 2H2O) under conditions which were found to be optimal for zinc phosphate solubilization. Under conditions of glucose at 300 mM C and alanine as the N source in the zinc phosphate-amended agar medium, one of the mycobionts produced new crystals, which were morphologically distinct from the original zinc phosphate crystals. It is concluded that medium composition influences the metal-phosphate solubilizing ability of ericoid mycobionts. The results are discussed in relation to the possible mechanisms involved in solubilization and the potential benefits of metal-phosphate solubilization to ericoid mycobionts and their host plants.