Bacterial colonization of the phyllosphere of nineteen plant species and antimicrobial activity of their leaf secondary metabolites against leaf associated bacteria

Summary. The scope of this work was to examine whether leaf constitutive secondary metabolites play a role in determining bacterial colonization of the phyllosphere. To this aim, we surveyed nineteen native or cultivated plant species that share a common bacterial pool in a North Mediterranean area, and estimated the size of total and ice nucleation active (INA) bacterial populations on their leaves. Large differences in the colonization of their phyllosphere were found; the population size of epiphytic bacteria ranged from 7.5 × 10² to 1 × 10⁶ CFU/g fresh weight, in eucalypt and celery, respectively. Species native in Mediterranean-type climate areas, particularly those belonging to the group of aromatic plants, are characterized by scarce presence of INA bacteria. The antibacterial activity of essential oils, surface phenolics and leaf tissue extracts was also estimated against the INA strains P. syringae and E. herbicola, isolated from two of these plant species. E. herbicola proved more sensitive than P. syringae. Of the species examined, oregano [Origanum vulgare L. subsp. hirtum (Link.) Ietswaart], an aromatic plant, had the highest antimicrobial activity, whereas six species showed no activity at all. Further experiments were performed with oregano and bean (Phaseolus vulgaris L.) that represent two extremes in their secondary metabolite content. Both plants were inoculated with P. syringae. By the end of incubation, the bacterial population on bean plants was about 100 times higher than that on oregano leaves. Scanning electron micrographs showed that bacterial growth on oregano leaves was confined to sites away from glandular hairs. Results from the bacterial colonization survey together with those from the toxicity tests showed that all species rich in antibacterial secondary metabolites harbored low leaf bacterial populations. These results provide substantial evidence that leaf secondary metabolites function as constitutive defense chemicals against microbial invasions. However, the fact that species with non- or moderately active leaf secondary metabolites are not always highly colonized suggests mediation of other unknown factors, the contribution of which requires further investigation.     

Abundance and diversity of the phyllosphere bacterial communities of Mediterranean perennial plants that differ in leaf chemistry

Summary.  We studied the epiphytic bacterial communities of the summer leaves of eight perennial species naturally occurring in a Mediterranean ecosystem. The species differ in essential-oil content (from rich in essential oil to non-producers) and composition, and also in life form (from herbaceous species to tall shrubs). We compared the epiphytic bacterial communities on the basis of (i) their abundance, (ii) their metabolic profile (derived by use of the BIOLOG Ecoplate system) and (iii) richness and diversity of substrates that they use, as a measure of functional diversity. Among all species, the aromatic Melissa officinalis was the most abundantly colonized. The bacterial communities on the leaves of the aromatic Myrtus communis, Calamintha nepeta and Melissa officinalis, and also of Cistus incanus catabolized all 31 substrates offered; those on the evergreen-sclerophyllous species, Arbutus unedo and Quercus coccifera, catabolized only 14 and 17 substrates, respectively. Carbohydrates were consistently used abundantly by all communities, whereas carboxylic acids were most variably used. On average, the group of aromatic plants scored higher regarding bacterial abundance, and richness and diversity of substrates used by the bacterial communities on their leaves; the lowest values for both substrate-use indices were recorded in A. unedo. Bacterial abundance or richness or diversity of substrates used did not vary with leaf oil content. Abundance was positively correlated with both substrate-use indices. Results support claims that the antimicrobial effects of essential oils are not exerted so much under natural conditions as reports based on biassays with pathogens usually show. Although essential oils play a part in the microbial colonization of the phyllosphere, it is not likely that inhibition of phyllosphere bacteria is essential oils’ primary role, at least in the Mediterranean environment.     

Diversity of cyanobacterial phylotypes in a Mediterranean drinking water reservoir (Marathonas, Greece)

The structure of the cyanobacterial community in a large drinking water reservoir (Marathonas, Greece) was investigated in October 2007 and September 2008. Cyanobacteria-specific primers were used for the PCR amplification of cyanobacterial 16S rDNAs from three water column sites and the water collection tank. In total, 199 clones were sequenced representing 52 unique cyanobacterial, including chloroplast-related, and 11 non-cyanobacterial phylotypes. All cyanobacterial phylotypes belonged to the order Chroococcales. Cluster analysis showed that the cyanobacterial communities in 2007 in the three water column sites showed high similarity between the stations and low diversity (H?=?1.17???1.44), due to the occurring common phylotypes, while all sites in 2008 had very low similarities between them and higher diversity (H?=?1.56???2.40). Some of the most abundant phylotypes were closely related (>98%) to members of the genus Gloeocapsa and a potentially toxin-producing strain of Microcystis aeruginosa. The non-cyanobacterial phylotypes were either unaffiliated or belonged to the Verrucomicrobia, and were related with sequences originating from lake water habitats.     

Effects of Grazing Pressure on Succession Process and Productivity of Old Fields on Mediterranean Islands

Poterium spinosum, competitive under conditions of overgrazing and fire, accounted for 70%–85% of the total plant cover in all sites, except Santorini, where participation of spiny species was very low. In the recently abandoned sites, annuals accounted for 50%–60% of the standing biomass, but constituted only a tiny fraction in those abandoned for 30 years or more. This was not paralleled by any remarkable increase of the total plant cover. In fact, the yearly produced green biomass decreased with the years after abandonment. The productivity of old fields abandoned for a long time is insufficient to support the grazing animals present. This shortfall, given the animals' requirements, is most dramatic in Symi. The nonrecovering vegetation in the old fields of these mountainous islands constitutes a severe environmental threat; remedial measures appear imperative.     

Need for recovery after emotional labor: Differential effects of daily deep and surface acting

This diary study examines the psychological processes that contribute to daily recovery from emotional labor by combining emotion regulation with work‐home resources theories. We hypothesized that overall perceptions of display rules relate positively to daily deep and surface acting. Daily surface acting was expected to relate positively to exhaustion and negatively to flow during work and consequently, to a higher need for recovery at the end of the workday. In contrast, daily deep acting was hypothesized to relate positively to flow and negatively to exhaustion and consequently, to a lower need for recovery at the end of the workday. In turn, need for recovery was expected to associate negatively to vigor at bedtime through reduced relaxation during leisure. Fifty Dutch and Polish employees first filled in a survey, and then a diary for five consecutive workdays, twice per day: at the end of the workday and before sleep. Multilevel path analyses largely supported these hypotheses suggesting that surface acting has unfavorable implications, whereas deep acting has favorable implications for daily well‐being at work and recovery after work.     

Derivation of ozone flux-yield relationships for lettuce: a key horticultural crop

Ozone flux-response relationships were derived for lettuce, employing a multiplicative approach to model the manner in which stomatal conductance is influenced by key environmental variables, using a dataset collected during field experimentation in Crete and yield-response relationships derived from parallel open-top chamber experiments. Regional agronomic practices were adopted throughout. Computed versus measured data revealed that the derived model explained 51% (P<0.001) of the observed variation in stomatal conductance. Concentration-based indices were compared with flux-based indices. Analyses revealed a significant relationship between accumulated stomatal ozone flux and yield employing flux threshold cut-offs up to 4 nmol m(-2) s(-1). Regressions employing very low or zero flux thresholds resulted in the strongest yield-flux relationships (explaining approximately 80% (P<0.05) of the variation in the dataset).     

Stimulation of soil microbial activity by essential oils

Summary. Although essential oils are well known antimicrobial agents, some microorganisms are activated by them and can use them as a carbon and energy source; this is the case for soil bacteria from Mediterranean ecosystems. We examined the assumption that soil microorganisms when offered with an essential oil, to which they had been previously exposed, would respond faster making immediate use of the newly added substrate. Origanum vulgare subsp. hirtum, Rosmarinus officinalis, Mentha spicata, and Coridothymus capitatus plants were collected and their essential oils isolated. Soil samples from the upper surface layer, beneath these aromatic plants, were also collected. All possible combinations of essential oils and soil samples were examined as well as the effect of the oil of R. officinalis and the non-indigenous, Lavandula angustifolia, on soil samples collected from cultivated fields. Soil respiration was used as a measure of the microbial activity. Oils (0.1 ml) were repeatedly added to the soil samples (150 g) and CO₂ release was measured every seven days. Essential oils differed in their chemical composition. In spite of that, they activated respiration of the different soil samples, even of those not previously exposed to essential oils, to a comparable degree. These results suggest that essential oils are used as a carbon and energy source by rather ubiquitously occurring soil microorganisms and provide evidence that they would not accumulate in the soil, if environmental conditions favour growth of these microorganisms.     

Forecasting daily pollen concentrations using data-driven modeling methods in Thessaloniki,Greece

Airborne pollen have been associated with allergic symptoms in sensitized individuals, having a direct impact on the overall quality of life of a considerable fraction of the population. Therefore, forecasting elevated airborne pollen concentrations and communicating this piece of information to the public are key issues in prophylaxis and safeguarding the quality of life of the overall population. In this study, we adopt a data-oriented approach in order to develop operational forecasting models (1–7 days ahead) of daily average airborne pollen concentrations of the highly allergenic taxa: Poaceae, Oleaceae and Urticaceae. The models are developed using a representative dataset consisting of pollen and meteorological time-series recorded during the years 1987–2002, in the city of Thessaloniki, Greece. The input variables (features) of the models have been optimized by making use of genetic algorithms, whereas we evaluate the performance of three algorithms: i) multi-Layer Perceptron, ii) support vector regression and iii) regression trees originating from distinct domains of Computational Intelligence (CI), and compare the resulting models with traditional multiple linear regression models. Results show the superiority of CI methods, especially when forecasting several days ahead, compared to traditional multiple linear regression models. Furthermore, the CI models complement each other, resulting to a combined model that performs better than each one separately. The overall performance ranges, in terms of the index of agreement, from 0.85 to 0.93 clearly suggesting the potential operational use of the models. The latter ones can be utilized in provision of personalized and on-time information services, which can improve quality of life of sensitized citizens.