Foliar nitrogen uptake in broadleaf evergreen Mediterranean forests: Fertilisation experiment with labelled nitrogen.

Atmospheric nitrogen (N) deposition in Mediterranean sclerophyllous forests of Holm oak (Quercus rotundifolia, Q. ilex) in Spain often exceeds empirical critical loads established for ecosystem conservation. There are still uncertainties on the capacity of canopy retention and uptake of the atmospheric N deposited of these forests. Studying and analysing all the forest nitrogen-cycle processes is essential to understand the potential effect of N deposition in these ecosystems. This study conducted a year-long short-term fertilisation experiment with labelled ammonium (15N-NH4) and nitrate (15N-NO3) to estimate foliar N absorption rates and assess the influence of leaf phenology and meteorological seasonal variations. Fertilising solutions were prepared to simulate low and high wet N deposition concentration, based on data reported from previous studies. Additionally, ecophysiological and meteorological measurements were collected to explore potential relationships between absorption rates, plant activity, and weather conditions. The results showed that Holm oak leaves were able to absorb both oxidised and reduced N compounds, with higher rates of NH4+ absorption. N recovery of both NH4+ and NO3- was higher in the low concentration treatments, suggesting reduced effectiveness of absorption as concentration increases. Foliar absorption rates were leaf-age dependent, with the highest values observed in young developing leaves. Foliar uptake showed seasonal changes with a clear reduction during the summer, linked to drought and dry weather conditions, and showing also smaller leaf net assimilation and stomatal conductance. During the rest of the year, foliar N absorption was not clearly associated to plant physiological activity but with environmental conditions. Our findings suggest that Holm oak canopies could absorb an important part of the incoming N deposition, but this process is compound, season and leaf phenology dependent. Further research is therefore needed to better understand and model this part of the N cycle.

Ozone and Temperature May Hinder Adaptive Capacity of Mediterranean Perennial Grasses to Future Global Change Scenarios.

Climate warming is recognized as a factor that threatens plant species in Mediterranean mountains. Tropospheric ozone (O3) should also be considered as another relevant stress factor for these ecosystems since current levels chronically exceed thresholds for plant protection in these areas. The main aim of the present study was to study the sensitivity of four Mediterranean perennial grasses to O3 and temperature based on plant growth, gas exchange parameters (photosynthesis-A, stomatal conductance-gs, and water use efficiency-WUE), and foliar macro- (N, K, Ca, Mg, P, and S) and micronutrients (B, Cu, Fe, Mn, Mo, and Zn) content. The selected species were grasses inhabiting different Mediterranean habitats from mountain-top to semi-arid grasslands. Plants were exposed to four O3 treatments in Open-Top chambers, ranging from preindustrial to above ambient levels, representing predicted future levels. Chamber-less plots were considered to study the effect of temperature increase. Despite the general tolerance of the grasses to O3 and temperature in terms of biomass growth, WUE and foliar nutrient composition were the most affected parameters. The grass species studied showed some degree of similarity in their response to temperature, more related with phylogeny than to their tolerance to drought. In some species, O3 or temperature stress resulted in low A or WUE, which can potentially hinder plant tolerance to climate change. The relationship between O3 and temperature effects on foliar nutrient composition and plant responses in terms of vegetative growth, A, gs, and WUE constitute a complex web of interactions that merits further study. In conclusion, both O3 and temperature might be modifying the adaptation capacity of Mediterranean perennial grass species to the global change. Air pollution should be considered among the driving favors of biodiversity changes in Mediterranean grassland habitats.

The Effects of Ozone on Visual Attraction Traits of Erodium paularense (Geraniaceae) Flowers: Modelled Perception by Insect Pollinators.

Ozone (O3) effects on the visual attraction traits (color, perception and area) of petals are described for Erodium paularense, an endangered plant species. Plants were exposed to three O3 treatments: charcoal-filtered air (CFA), ambient (NFA) and ambient + 40 nL L-1 O3 (FU+) in open-top chambers. Changes in color were measured by spectral reflectance, from which the anthocyanin reflectance index (ARI) was calculated. Petal spectral reflectance was mapped onto color spaces of bees, flies and butterflies for studying color changes as perceived by different pollinator guilds. Ozone-induced increases in petal reflectance and a rise in ARI under NFA were observed. Ambient O3 levels also induced a partial change in the color perception of flies, with the number of petals seen as blue increasing to 53% compared to only 24% in CFA. Butterflies also showed the ability to partially perceive petal color changes, differentiating some CFA petals from NFA and FU+ petals through changes in the excitation of the UV photoreceptor. Importantly, O3 reduced petal area by 19.8 and 25% in NFA and FU+ relative to CFA, respectively. In sensitive species O3 may affect visual attraction traits important for pollination, and spectral reflectance is proposed as a novel method for studying O3 effects on flower color.

DNA integrity and ecophysiological responses of Spanish populations of Ulmus glabra to increasing ozone levels.

Ulmus glabra is a deciduous tree with a wide distribution in the Eurosiberian region. The southernmost populations, in the Mediterranean area, are fragmented in mountain areas which act as a refugium. These small relict populations can act as sentinel of global change, including climate change and impacts of human activities such as air pollution. Besides, tropospheric ozone (O3) is an additional stress factor in the Mediterranean region affecting plant physiology and health. Moreover, oxidative stress caused by O3 could increase DNA damage in plants cells. U. glabra 4-year-old seedlings originated from a natural population growing in the Guadarrama mountain range (central Spain), were exposed in Open Top Chambers to four O3 treatments: charcoal filtered air, non-filtered air reproducing ambient levels, non-filtered air supplemented with 15 nl l-1 O3 and non- filtered air supplemented with 30 nl l-1 O3. Ozone effects on the DNA integrity through Comet assay were evaluated and eco-physiological responses were explored as well as. Comet assay showed a significant increase of DNA damage with increasing levels of O3 after only one-month exposure, when no eco-physiological symptoms of damage could be detected. Comet assay could thus be suggested as a predictive test to detect DNA damage induced in plants by other abiotic stresses as well as to identify tolerant and sensitive species or in preservation strategies of small relict populations. The discovery of a test for an early identification of stressed plants could be important to speed the selection of tolerant individuals for breeding programmes.