Morphoanatomical, Physiological, and Biochemical Indicators in Lactuca sativa L. Germination and Growth in Response to Fluoride.

Fluoride is one of the main phytotoxic environmental pollutants, and high concentrations (10-30 mg L-1) are commonly detected in surface and groundwater. Little, however, is known about the effects of this pollutant on crops that require irrigation during their development, which, in addition to phytotoxicity, may cause negative human health effects. Thus, the aim of this study was to characterize the effects of potassium fluoride (KF) on the germination of lettuce seeds and identify the physiological and anatomical markers of this pollutant's action on plants exposed to it during growth. Initially, lettuce seeds were sown in gerboxes and soaked in solutions containing 0 mg L-1, 10 mg L-1, 20 mg L-1, and 30 mg L-1 KF. Plants grown in a greenhouse were treated daily with KF irrigation at the same KF concentrations for 40 days. KF exposure reduced the germination rate and germination speed index of lettuce seeds at 20 mg L-1 and 30 mg L-1, resulting in compromised root development at the highest KF concentration. Lettuce plants displayed a slight photosynthesis reduction and a significant photochemical efficiency decrease after exposures to all KF concentrations. Lower chlorophyll contents and nitrogen balance indices were observed in plants exposed to 30 mg L-1 KF. On the other hand, increases in phenolic compounds and malondialdehyde were noted with increasing KF concentrations. Lettuce plants can, therefore, accumulate fluoride in leaves when irrigated with KF-rich water. The investigated physiological and biochemical variables were proven to be adequate fluoride action biomarkers in lettuce plants and may become an important tool in the study of olericulture contaminants.

Isotopic composition (δ13C and δ15N) in the soil-plant system of subtropical urban forests.

This study brings information on the dynamics of C and N in urban forests in a subtropical region. We tested the hypothesis that C and N isotopic sign of leaves and soil and physiological traits of trees would vary from center to periphery in a megacity, considering land uses, intensity of automotive fleet and microclimatic conditions. 800 trees from four fragments were randomly chosen. Soil samples were collected at every 10 cm in trenches up to 1 m depth to analyze C and N contents. Both, plants and soil were assessed for δ13C, δ15N, %C and %N. Physiological traits [carbon assimilation (A)], CO2 internal and external pressure ratio (Pi/Pa) and intrinsic water use efficiency iWUE were estimated from δ13C and Δ Î´13C in leaves and soil ranged from -27.42 ‰ to -35.39 ‰ and from -21.22 ‰ to -28.18 ‰, respectively, and did not vary along the areas. Center-periphery gradient was not evidenced by C. Emissions derived from fossil fuel and distinct land uses interfered at different levels in δ13C signature. δ15N in the canopy and soil varied clearly among urban forests, following center-periphery gradient. Leaf δ15N decreased from the nearest forest to the city center to the farthest, ranging from <3 ‰ to <-3 ‰. δ15N was a good indicator of atmospheric contamination by NOx emitted by vehicular fleet and a reliable predictor of land use change. %N followed the same trend of δ15N either for soils or leaves. Forest fragments located at the edges of the center-periphery gradient presented significantly lower A and Pi/Pa ratio and higher iWUE. These distinct physiological traits were attributed to successional stage and microclimatic conditions. Results suggest that ecosystem processes related to C and N and ecophysiological responses of urban forests vary according to land use and vehicular fleet.

Strategic roadmap to assess forest vulnerability under air pollution and climate change.

Although it is an integral part of global change, most of the research addressing the effects of climate change on forests have overlooked the role of environmental pollution. Similarly, most studies investigating the effects of air pollutants on forests have generally neglected the impacts of climate change. We review the current knowledge on combined air pollution and climate change effects on global forest ecosystems and identify several key research priorities as a roadmap for the future. Specifically, we recommend (1) the establishment of much denser array of monitoring sites, particularly in the South Hemisphere; (2) further integration of ground and satellite monitoring; (3) generation of flux-based standards and critical levels taking into account the sensitivity of dominant forest tree species; (4) long-term monitoring of N, S, P cycles and base cations deposition together at global scale; (5) intensification of experimental studies, addressing the combined effects of different abiotic factors on forests by assuring a better representation of taxonomic and functional diversity across the ~73,000 tree species on Earth; (6) more experimental focus on phenomics and genomics; (7) improved knowledge on key processes regulating the dynamics of radionuclides in forest systems; and (8) development of models integrating air pollution and climate change data from long-term monitoring programs.

Fluoride effect indicators in Phaseolus vulgaris seeds and seedlings.

Background: Fluoride (F) is one of the main environmental pollutants, and high concentrations are commonly detected in the air and in both surface and groundwater. However, the effects of this pollutant on seed germination and on the initial growth of crop seedlings are still poorly understood. In this context, the aim of this study was to assess morphoanatomical, physiological and biochemical fluoride effect indicators in Phaseolus vulgaris L. seeds and seedlings. Methods: P. vulgaris seeds were exposed to a liquid potassium fluoride solution (KF, pH 6.0) at concentrations of 0 (control), 10, 20, 30 mg L-1 for 7 days. A completely randomized experimental design was applied, consisting of four treatments with four replications each. During the experimental period, physiological (7 days) anatomical and histochemical (2 days), biochemical and chemical (4 days) assessments. An analysis of variance was performed followed by Dunnett's test. to determine significant differences between the KF-exposed groups and control seeds; and a multivariate analysis was performed. Results: The germination parameters, and anatomical, morphological, physiological, biochemical and nutritional characteristics of the seedlings did not show negative effects from exposure to KF at the lowest doses evaluated. On the other hand, treatment with the highest dose of KF (30 mg L-1) resulted in a lower germination rate index and increase in abnormal seedlings, and higher electrical conductivity. A lower root length, magnesium content and photochemical efficiency were also observed. The exposure of P. vulgaris to KF, regardless the dose did not affect seeds anatomy and the accumulation of starch and proteins, in relation to the control group. Conclusions: Our findings demonstrated that P. vulgaris seedlings were tolerant to KF solutions up to 20 mg L-1, and sensitive when exposed to 30 mg KF L-1.

Pah levels in the soil-litter-vegetation-atmosphere system of Atlantic Forest remnants in Southeast Brazil.

Although the Brazilian Atlantic Forest is a hotspot for biodiversity conservation, it is one of the most fragmented biomes in Brazil and also affected by air pollutants such as polycyclic aromatic hydrocarbons (PAHs). The study aimed at measuring the PAH levels in leaf trees, litter, soil, and atmosphere of two Atlantic Forest remnants impacted by air pollutants during summer and winter periods; identifying emission sources; and investigating the relationship among the PAH concentrations in the soil, litter, leaves, and atmosphere. Site 1 is situated in the largest South American city, with rainy summers and dry winters, and characterized by intense urbanization. Site 2 is situated in a large forest continuum and is characterized by wet climate with no defined dry seasons. It is more distant from the anthropogenic urban sources than site 1, but closer to an industrial complex. No differences were detected for PAH amounts (summer + winter) in the particles and wet deposition fluxes between sites. In site 1, the highest concentrations of PAHs in the particles were measured during the winter while in the leaf trees were measured during the summer. PMF model showed that sites 1 and 2 receive PAHs mainly from vehicle emissions and industrial activities, respectively. The accumulation of heavier compounds in soil and leaves via wet deposition was more evident in site 2. PAHs were mainly stored in the soil of site 1, contrasting with site 2, where they were retained in litter, which were attributed to disturbances of decomposer community and reduced decomposition rates.

Searching for biomarkers of early detection of 2,4-D effects in a native tree species from the Brazilian Cerrado biome.

Biodiversity in the Brazilian Cerrado biome has been declining sharply with the continued expansion of agriculture and the excessive use of herbicides. Thus, the aim of this study was to evaluate the morphophysiological and biochemical responses in Dipteryx alata plants to various doses of the herbicide 2,4-D. Specific biomarkers that characterize the phytoindicator potential of this species were determined. Gas exchange, chlorophyll a fluorescence, photosynthetic pigments, and the activities of antioxidant enzymes and cellulase were performed after 24, 96 and/or 396 hours after 2,4-D application (HAA). The herbicide caused higher antioxidant enzymatic activity 24 HAA and damage to the photosynthetic machinery after 96 HAA. Reduction in gas exchange, chlorophyll content, and photochemical traits were observed. Increased respiratory rates, non-photochemical quenching, and carotenoid concentrations in 2,4-D-treated plants were important mechanisms in the defense against the excess energy absorbed. Furthermore, the absence of leaf symptoms suggested tolerance of D. alata to 2,4-D. Nevertheless, changes in the photosynthetic and biochemical metabolism of D. alata are useful as early indicators of herbicide contamination, especially in the absence of visual symptoms. These results are important for early monitoring of plants in conserved areas and for preventing damage to sensitive species.

Metabolic and physiological alterations indicate that the tropical broadleaf tree Eugenia uniflora L. is sensitive to ozone.

Eugenia uniflora L. is an important fruit tree native to tropical South America that adapts to different habitats, thanks to its metabolic diversity and ability to adjust the leaf antioxidant metabolism. We hypothesized that this metabolic diversity would also enable E. uniflora to avoid oxidative damage and tolerate the enhanced ozone (O3) concentrations that have been registered in the (sub)tropics. We investigated whether carbohydrates, polyphenols and antioxidants are altered and markers of oxidative damage (ROS accumulation, alterations in leaf gas exchange, growth and biomass production) are detected in plants exposed to two levels of O3 (ambient air and twice elevated ozone level in a O3-FACE system for 75 days). Phytotoxic O3 dose above a threshold of 0 nmol m-2 s-1 (POD0) and accumulated exposure above 40 ppb (AOT40) were 3.6 mmol m-2 and 14.898 ppb h at ambient, and 4.7 mmol m-2 and 43.881 ppb h at elevated O3. Twenty-seven primary metabolites and 16 phenolic compounds were detected in the leaves. Contrary to the proposed hypothesis that tropical broadleaf trees are relatively O3 tolerant, we concluded that E. uniflora plants are sensitive to elevated O3 concentrations. Experimental POD0 values were lower than the critical levels for visible foliar O3, because of low stomatal conductance. In spite of this low stomatal O3 uptake, we found classic O3 injury, e.g. reduction in carbohydrates and fatty acids concentrations; non-significant changes in the polyphenol profile; inefficient antioxidant responses; increased contents of ROS and indicators of lipid peroxidation; reductions in stomatal conductance, net photosynthesis, root/shoot ratio and height growth. However, we also found some compensation mechanisms, e.g. increased leaf concentration of polyols for protecting the membranes, and increased leaf number for compensating the decline of photosynthetic rate. These results help filling the knowledge gap about tropical tree responses to O3.

Trace metals at the tree-litter-soil- interface in Brazilian Atlantic Forest plots surrounded by sources of air pollution.

Passive biomonitoring was applied in four Atlantic forest plots in southeast Brazil, affected by different levels of trace metal pollution (OP site located in Minas Gerais State and PEFI, PP and STG located in São Paulo State). Native tree species were selected as biomonitors according to their abundance in each plot and successional classification. Current trace metal concentrations in total suspended particles, leaves of non-pioneer (NPi) and pioneer (Pi) species, topsoil (0-20 cm) and litter and concentration ratios at the plant/soil interface were analyzed to verify the atmosphere-plant-soil interactions, basal concentrations, spatial variations and metal accumulation at the ecosystem level. Redundant analysis helped to identify similar characteristics of metal concentrations in PP and PEFI, which can be influenced by the high concentrations of elements related to anthropogenic inputs. Analysis of variance and multivariate statistics indicated that the trees of OP presented higher concentrations of Cr, Fe, Mn and Ni than those in the other sites. High enrichment of Cd, Fe, Ni in non-pioneer plants indicated that the PP forest (initially considered as the least polluted) has still been affected by metal pollution. Soil collected in STG was enriched by all elements, however these elements were low available for plant uptake. Metal deposited in leaves and litter was an important sink for soil cycling, nevertheless, these metals are not bioavailable in most cases. Non-pioneer tree species revealed to be more appropriate than pioneer species to indicate the current panorama of the contamination and bioavailability levels of trace metals in the tree community-litter-soil interface of the Atlantic forest remnants included in this study.

Ozone affects plant, insect, and soil microbial communities: A threat to terrestrial ecosystems and biodiversity.

Elevated tropospheric ozone concentrations induce adverse effects in plants. We reviewed how ozone affects (i) the composition and diversity of plant communities by affecting key physiological traits; (ii) foliar chemistry and the emission of volatiles, thereby affecting plant-plant competition, plant-insect interactions, and the composition of insect communities; and (iii) plant-soil-microbe interactions and the composition of soil communities by disrupting plant litterfall and altering root exudation, soil enzymatic activities, decomposition, and nutrient cycling. The community composition of soil microbes is consequently changed, and alpha diversity is often reduced. The effects depend on the environment and vary across space and time. We suggest that Atlantic islands in the Northern Hemisphere, the Mediterranean Basin, equatorial Africa, Ethiopia, the Indian coastline, the Himalayan region, southern Asia, and Japan have high endemic richness at high ozone risk by 2100.

Fluorescence emission spectra of target chloroplast metabolites (flavonoids, carotenoids, lipofuscins, pheophytins) as biomarkers of air pollutants and seasonal tropical climate.

Chloroplasts have luminescent metabolites-chlorophyll being the most known one-whose fluorescence emission may be a useful tool to assess the physiological status of the plant. Some antioxidants (flavonoids and carotenoids), and byproducts of membrane rupture (lipofuscins) and chlorophyll degradation (pheophytins), are chloroplasts' fluorescent metabolites directly involved in plant response to environmental stressors and pollutants and may act as a biomarker of stress. Here we hypothesized that climatic variations and air pollutants induce alterations in the emission profile of chloroplasts' fluorescent metabolites in Tillandsia usneoides (Bromeliaceae). To test this hypothesis, an active biomonitoring study was performed during 2 years in five polluted sites located at the Metropolitan Region of Campinas (São Paulo State, Brazil), aiming to identify target chloroplasts' fluorescent metabolites acting as biomarkers of environmental stress. In situ identification and quantification of the intensity of the fluorescence emission from target metabolites (flavonoids, carotenoids, lipofuscins, and pheophytins) were performed by the observation of fresh leaf sections under confocal laser scanning microscopy. Changes in the profile of fluorescence emission were correlated with local climate and air pollution data. The fluorescence emissions of flavonoids and carotenoids varied seasonally, with significant influence of rainfall and NO2. Our results expand the use of T. usneoides as a bioindicator by using alterations in the fluorescence emission profile of chloroplast metabolites. This application may be especially interesting for NO2 biomonitoring.

Dipteryx alata, a tree native to the Brazilian Cerrado, is sensitive to the herbicide nicosulfuron.

The expansion of land use for agricultural interests and the excessive use of herbicides are among the causes of biodiversity losses in the Brazilian Cerrado biome. Therefore, we aimed to test the hypothesis that Dipteryx alata Vogel, a common species in this biome, is sensitive to nicosulfuron because of its high phytotoxicity. We evaluated physiological, biochemical and morphological responses in D. alata plants exposed to increasing doses of the herbicide. Young plants were transplanted to 10 L pots containing substrate composed of soil and sand (2:1) after fertilization. After an acclimation period, the following doses of nicosulfuron were applied: 0 (control), 6, 12, 24, 48, and 60 g a.e. ha-1. The experiment was conducted in a randomized block design factorial scheme with six doses of nicosulfuron, three evaluation times, and five replicates per treatment. The effects of the herbicide were assessed by measuring gas exchange, chlorophyll a fluorescence, photosynthetic pigments, membrane permeability, antioxidant enzymes and acetolactate synthase. Nicosulfuron altered the photosynthetic machinery and enzymatic metabolism of D. alata. Reductions in physiological traits, increased catalase and ascorbate peroxidase activities, enhanced malondialdehyde concentrations rate of electrolyte leakage and decreased acetolactate synthase activity in response to nicosulfuron all suggest that D. alata is sensitive to this herbicide.

A quantitative assessment of hormetic responses of plants to ozone.

Evaluations of ozone effects on vegetation across the globe over the last seven decades have mostly incorporated exposure levels that were multi-fold the preindustrial concentrations. As such, global risk assessments and derivation of critical levels for protecting plants and food supplies were based on extrapolation from high to low exposure levels. These were developed in an era when it was thought that stress biology is framed around a linear dose-response. However, it has recently emerged that stress biology commonly displays non-linear, hormetic processes. The current biological understanding highlights that the strategy of extrapolating from high to low exposure levels may lead to biased estimates. Here, we analyzed a diverse sample of published empirical data of approximately 500 stimulatory, hormetic-like dose-responses induced by ozone in plants. The median value of the maximum stimulatory responses induced by elevated ozone was 124%, and commonly <150%, of the background response (control), independently of species and response variable. The maximum stimulatory response to ozone was similar among types of response variables and major plant species. It was also similar among clades, between herbaceous and woody plants, between deciduous and evergreen trees, and between annual and perennial herbaceous plants. There were modest differences in the stimulatory response between genera and between families which may reflect different experimental designs and conditions among studies. The responses varied significantly upon type of exposure system, with open-top chambers (OTCs) underestimating the maximum stimulatory response compared to free-air ozone-concentration enrichment (FACE) systems. These findings suggest that plants show a generalized hormetic stimulation by ozone which is constrained within certain limits of biological plasticity, being highly generalizable, evolutionarily based, and maintained over ecological scales. They further highlight that non-linear responses should be taken into account when assessing the ozone effects on plants.

Predicting ozone levels from climatic parameters and leaf traits of Bel-W3 tobacco variety.

Air pollution has been identified as a major cause of environmental and human health damage. O3 is an oxidative pollutant that causes leaf symptoms in sensitive plants. This study aims to adjust a multilinear model for the monitoring of O3 in subtropical climatic conditions by associating O3 concentrations with measurements of morphological leaf traits in tobacco plants and different environmental variables. The plants were distributed into five areas (residential, urban or industrial) in the southern region of Brazil and exposed during 14 periods, of 14 days each, during the years of 2014 and 2015. The environmental variables and leaf traits during the exposure periods were described by mean, median, standard deviation and minimum and maximum values. Spearman correlation and multiple linear regression analyses were applied on data from exposure periods. Leaf injury index, leaf area, leaf dry mass, temperature, relative humidity, global solar radiation and accumulated rainfall were used in the regression analyses to select the best models for predicting O3 concentrations. Leaf injury characteristically caused by O3 was verified in all areas and periods of plant exposure. Higher values of leaf injury (24.5% and 27.7%) were registered in the 13th and 12th exposure periods during spring and in areas influenced by urban and industrial clutches. The VPD, temperature, global solar radiation and O3 were correlated to leaf injury. Environmental variables [leaf area, leaf dry mass, global solar radiation and accumulated rainfall] and primarily the VPD were fundamental to improve the adjustments done in the bioindicator model (R2 ≥ 0.73). Our research shows that biomonitoring employing the tobacco "Bel-W3" can be improved by measuring morphological leaf traits and meteorological parameters. Additionally, O3 fumigation experiment should be performed with biomonitoring as conducted in this study, which are useful in understanding the role of other environmental factors.

The passion fruit liana (Passiflora edulis Sims, Passifloraceae) is tolerant to ozone.

Passiflora edulis Sims is a liana species of high economic interest and is an interesting model plant for understanding ozone action on disturbed vegetation. In this work we hypothesized that P. edulis has adaptive responses to oxidative stress that enable it to tolerate ozone damage based on its capacity to grow under a diversity of environmental conditions and to dominate disturbed areas. We exposed seedlings to three levels of ozone in a Free-Air Controlled Exposure (FACE) system (22, 41 and 58 ppb h AOT40 and 13.52, 17.24 and 20.62 mmol m-2 POD0, over 97 days) for identifying its tolerance mechanisms. Anatomical (leaf blade structure and fluorescence emission of chloroplast metabolites), physiological (leaf gas exchange, growth rate and biomass production) and biochemical (pigments, total sugars, starch, enzymatic and non-enzymatic antioxidant metabolites, reactive oxygen species and lipid peroxidation derivatives) responses were assessed. Ozone caused decreased total number of leaves, hyperplasia and hypertrophy of the mesophyll cells, and accelerated leaf senescence. However, O3 did not affect carbohydrates content, net photosynthetic rate, or total biomass production, indicating that the carboxylation efficiency and associated physiological processes were not affected. In addition, P. edulis showed higher leaf contents of ascorbic acid, glutathione (as well high ratio between their reduced and total forms), carotenoids, and flavonoids located in the chloroplast outer envelope membrane. Our results indicate that P. edulis is an O3-tolerant species due to morphological acclimation responses and an effective antioxidant defense system represented by non-enzymatic antioxidants, which maintained the cellular redox balance under ozone.

Commentary: EPA's proposed expansion of dose-response analysis is a positive step towards improving its ecological risk assessment.

The United States Environmental Protection Agency (US EPA) has recently proposed changes to strengthen the transparency of its pivotal regulatory science policy and procedures. In this context, the US EPA aims to enhance the transparency of dose-response data and models, proposing to consider for the first time non-linear biphasic dose-response models. While the proposed changes have the potential to lead to markedly improved ecological risk assessment compared to past and current approaches, we believe there remain open issues for improving the quality of ecological risk assessment, such as the consideration of adaptive, dynamic and interactive effects. Improved risk assessment including adaptive and dynamic non-linear models (beyond classic threshold models) can enhance the quality of regulatory decisions and the protection of ecological health. We suggest that other countries consider adopting a similar scientific-regulatory posture with respect to dose-response modeling via the inclusion of non-linear biphasic models, that incorporate the dynamic potential of biological systems to adapt (i.e., enhancing positive biological endpoints) or maladapt to low levels of stressor agents.

Efficiency of biomonitoring methods applying tropical bioindicator plants for assessing the phytoxicity of the air pollutants in SE, Brazil.

In the tropical region, the greatest challenge of the biomonitoring approach is to establish linear relationships between biomarkers measured in plants and pollutant concentrations, since the bioindicator responses can be intensified or restricted by climatic variations. In southeastern Brazil, there are two regions affected by air pollution, where the Atlantic Forest remains and should be preserved. Consequently, both areas have been monitored by biomonitoring procedures using standardized and tropical plants. The industrial complex settled in Cubatão is one of the world's most famous examples of environmental pollution and degradation, with consequent decline of the Atlantic Forest. An oil refinery is among the most polluting industries in the Cubatão region. The other region is located in the Metropolitan Region of Campinas (MRC). The MRC has been affected by high levels of air pollutants originated from road traffic and is responsible for over 80% of CO, NOx, and hydrocarbon emissions and develops industrial activities that emit about 70% of the particulate matter present in the region. Both regions are distinguished by the climate, despite the fact that they are only about 130 km far from each other. Several studies carried out by our group in these regions aimed to establish the best native tree species and respective potential biomarkers for future assessment of pollution effects on tropical Forests. We present a critical review about the efficiency of native species compared to standardized bioindicator plants considering antioxidant defense system, nutrient accumulation, and microscopic aspects when exposed to atmospheric pollutants and climate.

Oxidant-antioxidant balance and tolerance against oxidative stress in pioneer and non-pioneer tree species from the remaining Atlantic Forest.

The extensive land occupation in Southeast Brazil has resulted in climatic disturbances and environmental contamination by air pollutants, threatening the Atlantic forest remnants that still exist in that region. Based on previous results, we assumed that pioneer tree species are potentially more tolerant against environmental oxidative stress than non-pioneer tree species from that Brazilian biome. We also assumed that reactive oxygen species (ROS) are accumulated in higher proportions in leaves of non-pioneer trees, resulting in changes in the oxidant-antioxidant balance and in more severe oxidative damage at the cellular level than in the leaves of pioneer trees. We tested these hypotheses by establishing the relationship between oxidants (ROS), changes in key antioxidants (among enzymatic and non-enzymatic compounds) and in a lipid peroxidation derivative in their leaves, as well as between ROS accumulation and oscillations in environmental stressors, thus permitting to discuss comparatively for the first time the oxidant-antioxidant balance and the tolerance capacity of tree species of the Atlantic Forest in SE Brazil. We confirmed that the non-pioneer tree species accumulated higher amounts of superoxide and hydrogen peroxide in palisade parenchyma and epidermis, showing a less effective antioxidant metabolism than the pioneer species. However, the non-pioneer species showed differing capacities to compensate the oxidative stress in both years of study, which appeared to be associated with the level of ROS accumulation, which was evidently higher in 2015 than in 2016. We also applied exploratory multivariate statistics, which revealed that the oscillations in these biochemical leaf responses in both functional groups coincided with the oscillations in both climatic conditions and air pollutants, seemingly showing that they had acclimated to the stressful oxidative environment observed and may perpetuate in the disturbed forest remnants located in SE Brazil.

Morphoanatomical and physiological changes in Bauhinia variegata L. as indicators of herbicide diuron action.

The wide use of the herbicide diuron has compromised surrounding uncultivated areas, resulting in acute and/or chronic damage to non-target plants. Thus, the aim of this research was to evaluate physiological and morphoanatomical responses in Bauhinia variegata L. plants to different doses of diuron. Seedlings of 90-day-old B. variegata were transplanted into 10liter pots. After an acclimation period (about 30 days), treatments consisting of different diuron doses were applied: 0 (control), 400, 800, 1600, and 2400g ai ha-1. The experiment was conducted in a randomized block design in a 5×5 factorial scheme with five doses of diuron five evaluation times, and five replicates per treatment. Anatomical and physiological injuries were observed in leaves of Bauhina variegata 10h after diuron application. Disruption of waxes was observed on both sides of the leaves of plants exposed since the lowest dose. Plasmolysis in cells were observed in treated leaves; more severe damage was observed in plants exposed to higher doses, resulting in rupture of epidermis. The diuron herbicide also caused gradual reduction in the gas exchange and chlorophyll fluorescence variables. Among the morphoanatomical and physiological variables analyzed, the non-invasive ones (e.g., ETR, YII, and Fv/Fm) may be used as biomarkers of diuron action in association with visible symptoms. In addition, changes in leaf blade waxes and chlorophyll parenchyma damage may also be considered additional leaf biomarkers of diuron herbicide action.

Tillandsia usneoides: A successful alternative for biomonitoring changes in air quality due to a new highway in São Paulo, Brazil.

Tillandsia usneoides is an aerial epiphytic bromeliad that absorbs water and nutrients directly from the atmosphere by scales covering its surface. We expanded the use of this species as a broader biomonitor based on chemical and structural markers to detect changes in air quality. The usefulness of such comprehensive approach was tested during the construction and opening of a highway (SP-21) in São Paulo State, Brazil. The biomonitoring study was performed from 2009 to 2012, thus comprising the period during construction and after the highway inauguration. Metal accumulation and structural alterations were assessed, in addition to microscopy analyses to understand the metal chelation in plant tissues and to assess the causes of alterations in the number and shape of scale cells. Altogether, our analyses support the use of this species as a wide biomonitor of air quality in urbanized areas.