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.

Functional Leaf Traits and Diurnal Dynamics of Photosynthetic Parameters Predict the Behavior of Grapevine Varieties Towards Ozone.

A comparative study on functional leaf treats and the diurnal dynamics of photosynthetic processes was conducted on 2-year-old potted plants of two grape (Vitis vinifera L.) varieties (Aleatico, ALE, and Trebbiano giallo, TRE), exposed under controlled conditions to realistic concentrations of the pollutant gas ozone (80 ppb for 5 h day(-1), 8:00-13:00 h, + 40 ppb for 5 h day(-1), 13:00-18:00 h). At constitutive levels, the morphological functional traits of TRE improved leaf resistance to gas exchange, suggesting that TRE is characterized by a potential high degree of tolerance to ozone. At the end of the treatment, both varieties showed typical visible injuries on fully expanded leaves and a marked alteration in the diurnal pattern of photosynthetic activity. This was mainly due to a decreased stomatal conductance (-27 and -29% in ALE and TRE, in terms of daily values in comparison to controls) and to a reduced mesophyllic functioning (+33 and +16% of the intercellular carbon dioxide concentration). Although the genotypic variability of grape regulates the response to oxidative stress, similar detoxification processes were activated, such as an increased content of total carotenoids (+64 and +30%, in ALE and TRE), enhanced efficiency of thermal energy dissipation within photosystem II (+32 and +20%) closely correlated with the increased de-epoxidation index (+26 and +22%) and variations in content of some osmolytes. In summary, we can conclude that: the daily photosynthetic performance of grapevine leaves was affected by a realistic exposure to ozone. In addition, the gas exchange and chlorophyll a fluorescence measurements revealed a different quali-quantitative response in the two varieties. The genotypic variability of V. vinifera and the functional leaf traits would seem to regulate the acclimatory response to oxidative stress and the degree of tolerance to ozone. Similar photoprotective mechanisms were activated in the two varieties, though to a different extent.

Large-scale patterns of genetic variation in a female-biased dispersing passerine: the importance of sex-based analyses.

Dispersal affects the distribution, dynamics and genetic structure of natural populations, and can be significantly different between sexes. However, literature records dealing with the dispersal of migratory birds are scarce, as migratory behaviour can notably complicate the study of dispersal. We used the barn swallow Hirundo rustica as model taxon to investigate patterns of genetic variability in males and in females of a migratory species showing sex-biased dispersal. We collected blood samples (n = 186) over the period 2006 to 2011 from adults (H. r. rustica subspecies) nesting in the same breeding site at either high (Ireland, Germany and Russia) or low (Spain, Italy and Cyprus) latitude across Europe. We amplified the Chromo Helicase DNA gene in all birds in order to warrant a sex-balanced sample size (92 males, 94 females). We investigated both uniparental (mitochondrial ND2 gene) and biparental (microsatellite DNA: 10 loci) genetic systems. The mtDNA provided evidence for demographic expansion yet no significant partition of the genetic variability was disclosed. Nevertheless, a comparatively distant Russian population investigated in another study, whose sequences were included in the present dataset, significantly diverged from all other ones. Different to previous studies, microsatellites highlighted remarkable genetic structure among the studied populations, and pointed to the occurrence of differences between male and female barn swallows. We produced evidence for non-random patterns of gene flow among barn swallow populations probably mediated by female natal dispersal, and we found significant variability in the philopatry of males of different populations. Our data emphasize the importance of taking into account the sex of sampled individuals in order to obtain reliable inferences on species characterized by different patterns of dispersal between males and females.

How sensitive is Melissa officinalis to realistic ozone concentrations?

Lemon balm (Melissa officinalis, L.; Lamiaceae) was exposed to realistic ozone (O3) dosages (80 ppb for 5 h), because high background levels of O3 are considered to be as harmful as episodic O3 regimes. Temporal alterations of different ecophysiological, biochemical and structural parameters were investigated in order to test if this species can be considered as an O3-bioindicator regarding changes in background concentrations. At the end of ozone exposure, the plants did not exhibit any visible foliar symptoms, as only at microscopic level a small number of dead cells were found. Photosynthetic processes, however, were significantly affected. During and after the treatment, ozone induced a reduction in CO2 fixation capacity (up to 52% after 12 h from the beginning of the treatment) due to mesophyllic limitations. Intercellular CO2 concentration significantly increased in comparison to controls (+90% at the end of the post-fumigation period). Furthermore impairment of carboxylation efficiency (-71% at the end of the post-fumigation period compared to controls in filtered air) and membrane damage in terms of integrity (as demonstrated by a significant rise in solute leakage) were observed. A regulatory adjustment of photosynthetic processes was highlighted during the post-fumigation period by the higher values of qNP and (1-q(P)) and therefore suggests a tendency to reduce the light energy used in photochemistry at the expense of the capacity to dissipate the excess as excitation energy. In addition, the chlorophyll a/b ratio and the de-epoxidation index increased, showing a rearrangement of the pigment composition of the photosynthetic apparatus and a marked activation of photoprotective mechanisms.