Assessment of the effects of ozone exposure and plant competition on the reproductive ability of three therophytic clover species from Iberian pastures

Ozone (O3) phytototoxicity has been reported on a wide range of crops and wild Central European plantspecies, however no information has been provided regarding the sensitivity of plantspecies from dehesa Mediterranean therophytic grasslands in spite of their great plantspecies richness and the high O3 levels that are recorded in this area. A study was carried out in open-top chambers (OTCs) to assess the effects of O3 and competition on the reproductiveability of threecloverspecies: Trifolium cherleri, Trifolium subterraneum and Trifolium striatum. A phytometer approach was followed, therefore plants of these species were grown in mesoscosms composed of monocultures of four plants of each species, of threeplants of each species competing against a Briza maxima individual or of a single plant of each cloverspecies competing with threeB. maximaplants. Three O3 treatments were adopted: charcoal filtered air (CFA), non-filtered air (NFA) and non-filtered air supplemented with 40 nl l−1 of O3 (NFA+). The different mesocosms were exposed to the different O3 treatments for 45 days and then they remained in the open. Ozoneexposure caused reductions in the flower biomass of the threecloverspecies assessed. In the case of T. cherleri and T. subterraneum this effect was found following their exposure to the different O3 treatments during their vegetative period. An attenuation of these effects was found when the plants remained in the open. Ozone-induced detrimental effects on the seed output of T. striatum were also observed. The flower biomass of the cloverplants grown in monocultures was greater than when competing with one or threeB. maxima individuals. An increased flower biomass was found in the CFA monoculture mesocosms of T. cherleri when compared with the remaining mesocosms, once the plants were exposed in the open for 60 days. The implications of these effects on the performance of dehesa acid grasslands and for the definition of O3 critical levels is discussed

Assessment of the ozone sensitivity of 22 native plant species from mediterranean annual pastures based on visible injury

Ozone (O3) phytototoxicity has been reported on a wide range of plantspecies, inducing the appearance of specific foliar injury or increasing leaf senescence. No information regarding the sensitivity of plantspecies from dehesa Mediterranean grasslands has been provided in spite of their great biological diversity. A screening study was carried out in open-top chambers (OTCs) to assess the O3-sensitivity of 22 representative therophytes of these ecosystems based on the appearance and extent of foliar injury. A distinction was made between specific O3injury and non-specific discolorations. Three O3 treatments (charcoal-filtered air, non-filtered air and non-filtered air supplemented with 40 nl l−1 O3 during 5 days per week) and three OTCs per treatment were used. The Papilionaceae species were more sensitive to O3 than the Poaceae species involved in the experiment since ambient levels induced foliar symptoms in 67% and 27%, respectively, of both plant families. An O3-sensitivity ranking of the species involved in the assessment is provided, which could be useful for bioindication programmes in Mediterranean areas. The assessed Trifoliumspecies were particularly sensitive since foliar symptoms were apparent in association with O3 accumulated exposures well below the current critical level for the prevention of this kind of effect. The exposure indices involving lower cut-off values (i.e. 30 nl l−1) were best related with the extent of O3-induced injury on these species.

Modelling ozone stomatal flux over wheat under Mediterranean conditions

Correct estimation of leaf-level stomatal conductance (gsto) is central for current ozone (O3) risk assessment of wheat yield loss based on the absorbed O3 phytotoxic dose (POD). The gsto model parameterizations developed in Europe must be checked in the different climatic regions where they are going to be applied in order to reduce the uncertainties associated with the POD approach. This work proposes a new gsto model parameterization for estimating POD of Triticum aestivum and Triticum durum under Mediterranean conditions, based on phenological observations over 25 years and gsto field measurements during 5 growing seasons. Results show that POD in the Mediterranean area might be higher than previously estimated. However, caution must be paid when assessing the risk of yield loss for wheat in this area since field validation of O3 impacts is still limited.

Urban trees and atmospheric pollutants in big cities: Effects in Madrid

Una investigación sobre la mejora de la contaminación del aire (CA) por medio de arbolado urbano se realizó en Madrid, una ciudad con casi 4 M de habitantes, 2,8 M de vehículos y casi 3 M de árboles de mantenimiento público. La mayoría de los árboles estaban en dos bosques periurbanos. Los 650.000 restantes era pies de alineación y parques. Los taxones estudiados fueron Platanus orientalis (97.205 árboles), Ulmus sp. (70.557), Pinus pinea (49.038), Aesculus hippocastanum (22.266), Cedrus sp. (13.678) y Quercus ilex (1.650), de calles y parques. Muestras foliares se analizaron en diferentes épocas del año, así como datos de contaminación por PM10 de 28 estaciones de medición de la contaminación durante 30 años, y también la intensidad del tráfico (IMD) en 2.660 calles. La acumulación de metales pesados (MP) sobre hojas y dentro de estas se estimó en relación con la CA y del suelo y la IMD del tráfico. La concentración media de Ba, Cd, Cr, Cu, Mn, Ni, Pb y Zn en suelo (materia seca) alcanzó: 489,5, 0,7, 49,4, 60,9, 460,9, 12,8, 155,9 y 190,3 mg kg-1 respectivamente. Los árboles urbanos, particularmente coníferas (debido a la mayor CA en invierno) contribuyen significativamente a mejorar la CA sobre todo en calles con alta IMD. La capacidad de las seis sp. para capturar partículas de polvo en su superficies foliares está relacionada con la IMD del tráfico y se estimó en 16,8 kg/año de MP tóxicos. Pb y Zn resultaron ser buenos marcadores antrópicos en la ciudad en relación con el tráfico, que fue la principal fuente de contaminación en los árboles y suelos de Madrid. Las especies de árboles variaron en función de su capacidad para capturar partículas (dependiendo de las propiedades de sus superficies foliares) y acumular los MP absorbidos de los suelos. Las concentraciones foliares de Pb y Zn estuvieron por encima de los límites establecidos en diferentes sitios de la ciudad. La microlocalización de Zn mediante microscópico mostró la translocación al xilema y floema. Se detectaron puntos de contaminación puntual de Cu and Cr en antiguos polígonos industriales y la distribución espacial de los MP en los suelos de Madrid mostró que en incluso en zonas interiores del El Retiro había ciertos niveles elevados de [Pb] en suelo, tal vez por el emplazamiento la Real Fábrica de Porcelana en la misma zona hace 200 años. Distintas áreas del centro de la ciudad también alcanzaron niveles altos de [Pb] en suelo. Según los resultados, el empleo de una combinación de Pinus pinea con un estrato intermedio de Ulmus sp. y Cedrus sp. puede ser la mejor recomendación como filtro verde eficiente. El efecto del ozono (O3) sobre el arbolado en Madrid fue también objeto de este estudio. A pesar de la reducción de precursores aplicada en muchos países industrializados, O3 sigue siendo la principal causa de CA en el hemisferio norte, con el aumento de [O3] de fondo. Las mayores [O3] se alcanzaron en regiones mediterráneas, donde el efecto sobre la vegetación natural es compensado por el xeromorfismo y la baja conductancia estomática en respuesta los episodios de sequía estival característicos de este clima. Durante una campaña de monitoreo, se identificaron daños abióticos en hojas de encina parecidos a los de O3 que estaban plantadas en una franja de césped con riego del centro de Madrid. Dada la poca evidencia disponible de los síntomas de O3 en frondosas perennifolias, se hizo un estudio que trató de 1) confirman el diagnóstico de daño de O3, 2) investigar el grado de los síntomas en encinas y 3) analizar los factores ambientales que contribuyeron a los daños por O3, en particular en lo relacionado con el riego. Se analizaron los marcadores macro y micromorfológicos de estrés por O3, utilizando las mencionadas encinas a modo de parcela experimental. Los síntomas consistieron en punteado intercostal del haz, que aumentó con la edad. Además de un punteado subyacente, donde las células superiores del mesófilo mostraron reacciones características de daños por O3. Las células próximas a las zonas dañadas, presentaron marcadores adicionales de estrés oxidativo. Estos marcadores morfológicos y micromorfológicos de estrés por O3 fueron similares a otras frondosas caducifolias con daños por O3. Sin embargo, en nuestro caso el punteado fue evidente con AOT40 de 21 ppm•h, asociada a riego. Análisis posteriores mostraron que los árboles con riego aumentaron su conductancia estomática, con aumento de senescencia, manteniéndose sin cambios sus características xeromórficas foliares. Estos hallazgos ponen de relieve el papel primordial de la disponibilidad de agua frente a las características xeromórficas a la hora de manifestarse los síntomas en las células por daños de O3 en encina. ABSTRACT Research about air pollution mitigation by urban trees was conducted in Madrid (Spain), a southern European city with almost 4 M inhabitants, 2.8 M daily vehicles and 3 M trees under public maintenance. Most trees were located in two urban forests, while 650'000 trees along urban streets and in parks. The urban taxa included Platanus orientalis (97'205 trees), Ulmus sp. (70’557), Pinus pinea (49'038), Aesculus hippocastanum (22’266), Cedrus sp. (13'678 and Quercus ilex (1'650) along streets and parks. Leave samples were analysed sequentially in different seasons, PM10 data from 28 air monitoring stations during 30 years and traffic density estimated from 2’660 streets. Heavy metal (HM) accumulation on the leaf surface and within leaves was estimated per tree related to air and soil pollution, and traffic intensity. Mean concentration of Ba, Cd, Cr, Cu, Mn, Ni, Pb and Zn in topsoil samples (dry mass) amounted in Madrid: 489.5, 0.7, 49.4, 60.9, 460.9, 12.8, 155.9 and 190.3 mg kg-1 respectively. Urban trees, particularly conifers (due to higher pollution in winter) contributed significantly to alleviate air pollution especially near to high ADT roads. The capacity of the six urban street trees species to capture air-born dust on the foliage surface as related to traffic intensity was estimated to 16.8 kg of noxious metals from exhausts per year. Pb and Zn pointed to be tracers of anthropic activity in the city with vehicle traffic as the main source of diffuse pollution on trees and soils. Tree species differed by their capacity to capture air-borne dust (by different leaf surface properties) and to allocate HM from soils. Pb and Zn concentrations in the foliage were above limits in different urban sites and microscopic Zn revelation showed translocation in xylem and phloem tissue. Punctual contamination in soils by Cu and Cr was identified in former industrial areas and spatial trace element mapping showed for central Retiro Park certain high values of [Pb] in soils even related to a Royal pottery 200 years ago. Different areas in the city centre also reached high levels [Pb] in soils. According to the results, a combination of Pinus pinea with understorey Ulmus sp. and Cedrus sp. layers can be recommended for the best air filter efficiency. The effects of ozone (O3) on trees in different areas of Madrid were also part of this study. Despite abatement programs of precursors implemented in many industrialized countries, ozone remained the main air pollutant throughout the northern hemisphere with background [O3] increasing. Some of the highest ozone concentrations were measured in regions with a Mediterranean climate but the effect on the natural vegetation is alleviated by low stomatal uptake and frequent leaf xeromorphy in response to summer drought episodes characteristic of this climate. During a bioindication survey, abiotic O3-like injury was identified in foliage. Trees were growing on an irrigated lawn strip in the centre of Madrid. Given the little structural evidence available for O3 symptoms in broadleaved evergreen species, a study was undertaken in 2007 with the following objectives 1) confirm the diagnosis, 2) investigate the extent of symptoms in holm oaks growing in Madrid and 3) analyse the environmental factors contributing to O3 injury, particularly, the site water supply. Therefore, macro- and micromorphological markers of O3 stress were analysed, using the aforementioned lawn strip as an intensive study site. Symptoms consisted of adaxial and intercostal stippling increasing with leaf age. Underlying stippling, cells in the upper mesophyll showed HR-like reactions typical of ozone stress. The surrounding cells showed further oxidative stress markers. These morphological and micromorphological markers of ozone stress were similar to those recorded in deciduous broadleaved species. However, stippling became obvious already at an AOT40 of 21 ppm•h and was primarily found at irrigated sites. Subsequent analyses showed that irrigated trees had their stomatal conductance increased and leaf life-span reduced whereas their leaf xeromorphy remained unchanged. These findings suggest a central role of water availability versus leaf xeromorphy for ozone symptom expression by cell injury in holm oak.

Current ozone levels threaten gross primary production and yield of Mediterranean annual pastures and nitrogen modulates the response

Pastures are among the most important ecosystems in Europe considering their biodiversity and dis- tribution area. However, their response to increasing tropospheric ozone (O 3 ) and nitrogen (N) deposi- tion, two of the main drivers of global change, is still uncertain. A new Open-Top Chamber (OTC) experiment was performed in central Spain, aiming to study annual pasture response to O 3 and N in close to natural growing conditions. A mixture of six species of three representative families was sowed in the fi eld. Plants were exposed for 40 days to four O 3 treatments: fi ltered air, non- fi ltered air (NFA) repro- ducing ambient levels and NFA supplemented with 20 and 40 nl l � 1 O 3 . Three N treatments were considered to reach the N integrated doses of “ background ” , þ 20 or þ 40 kg N ha � 1 . Ozone signi fi cantly reduced green and total aboveground biomass (maximum reduction 25%) and increased the senescent biomass (maximum increase 40%). Accordingly, O 3 decreased community Gross Primary Production due to both a global reduction of ecosystem CO 2 exchange and an increase of ecosystem respiration. Nitrogen could partially counterbalance O 3 effects on aboveground biomass when the levels of O 3 were moderate, but at the same time O 3 exposure reduced the fertilization effect of higher N availability. Therefore, O 3 must be considered as a stress factor for annual pastures in the Mediterranean areas.