Using native plants to evaluate urban metal pollution and appoint emission sources in the Brazilian Steel Valley region.

In southeastern Brazil, the city of Ipatinga is inserted in the Steel Valley Metropolitan Region, which hosts the largest industrial complex for flat-steel production in Latin America, while also having one of the largest vehicle fleets in the entire country. Since potentially toxic elements (PTEs) are not emitted solely by industries, yet also by vehicular activity, the predominant emission source can be determined by evaluating the ratio between different elements, which are called technogenic tracers. We performed a biomonitoring assay using two tropical legumes, Paubrasilia echinata and Libidibia ferrea var. leiostachya, aiming to assess chemical markers for the origin of emissions in the region, distinguishing between different anthropogenic sources. Plants were exposed for 90 days in four urban sites and in a neighboring park which served as reference. After the experimental period, plants were evaluated for trace-metal accumulation. L. ferrea var. leiostachya retained lower amounts of metals associated with vehicular and industrial emission. The opposite was found with P. echinata, a species which should be recommended for biomonitoring of air pollution as a bioaccumulator. Plants of P. echinata were enriched with Fe, Al, Ni, Cr, and Ba, whereas plants of L. ferrea var. leiostachya were enriched with Fe, Cu, and Co. In both species, Fe was the element with which plants were enriched the most. Plants showed highest iron enrichment at Bom Retiro, the site downwind to the steel industry, which has shown to be the main particle emission source in the region.

Why is Brachiaria decumbens Stapf. a common species in the mining tailings of the Fundão dam in Minas Gerais, Brazil?

Currently, more than five years after the Fundão dam failure in Mariana, Minas Gerais, Brazil, Brachiaria decumbens Stapf. is the main grass in pasturelands affected by the mining tailings. The aim of this study was to investigate the reason for this fact as well as to determine the ecophysiological effects of mining tailings on B. decumbens and to test whether mixing the tailings with unaffected local soil enhances the affected soil properties. For the experiment, two different soils were collected, one unaffected soil without mining tailings (Ref) and the mining tailings (Tec), and we also created a mixture with 50 % of each soil type (Ref/Tec). We cultivated B. decumbens in the three soil treatments in a greenhouse for 110 days and evaluated soil physical-chemical properties and plant ecophysiology. Our results show that the tailings (Tec) compromised the normal ecophysiological state of B. decumbens. The species survived these adverse conditions due to its great efficiency in acquiring some elements. The soil management tested by this work mitigated the stress caused by tailings and can represent an alternative for the environmental recovery of the affected soils.

Effects of fluoride emission on the morphoanatomy of three plant species endemics to Brazil using passive biomonitoring.

Fluoride is the most phytotoxic atmospheric pollutant. The objective of this study was to evaluate the effects of fluoride emissions by an aluminum smelter on three plant species endemics to Brazil, located at Parque Estadual do Itacolomi (PEI). The monitored species were Byrsonima variabilis (Malpighiaceae), Myrceugenia alpigena (Myrtaceae), and Eremanthus erythropappus (Asteraceae), which were monitored during 9 months using passive biomonitoring at five different locations with different distances from the smelter. The monitored species did not show macroscopic phytotoxicity damage to fluoride; however, they did show microscopic damage. The species closer to the smelter presented more severe anatomic damages, such as rupture of cell walls, protoplast retraction, and trichome alterations. Damaged stomatal ledges, flaking epicuticular wax, and damages to trichomes were observed. M. alpigena showed a higher accumulation of fluoride than the other species at all monitored sites. The test for cell death with Evans Blue was positive for the three studied species. Through biomonitoring in the PEI, we concluded that the emissions from the aluminum smelter affect the native vegetation and that due to the greater accumulation of fluoride and the diversity of microscopic damage in M. alpigena, the use of this species in the monitoring of environments polluted by fluoride is enhanced.

Physiological and morphoanatomical effects of glyphosate in Eugenia uniflora, a Brazilian plant species native to the Atlantic Forest biome.

The herbicide glyphosate can cause severe ecotoxicological effects on non-target organisms. Eugenia uniflora L. (Myrtaceae) is very important for in situ environmental biomonitoring due to its wide distribution in the Atlantic Forest biome. Thus, this study aimed to evaluate the response of E. uniflora leaves to glyphosate. Eight-month-old plants were exposed to an aerial application of the herbicide at concentrations of 0, 144, 432, 864, and 1440 g a. e. ha-1 (grams of acid equivalent per hectare). Evaluations were performed on the 12th day after the glyphosate application (DAA). An accumulation of shikimic acid in the leaves of E. uniflora was observed. Glyphosate altered the photosynthetic parameters of the treated plants, with a drastic decrease in the photosynthetic rate, stomatal conductance, transpiration, and pigment content. There was an increase in Ci/Ca, lipid peroxidation, and electrolyte extravasation levels. Glyphosate also promoted ultrastructural, anatomical and visible damage to the E. uniflora leaves. Our findings indicate that glyphosate is phytotoxic to the native species E. uniflora at the tested doses. The presence of visible damage suggests that E. uniflora has remarkable potential as a bioindicator of glyphosate in the environment, making it a possible species for future biomonitoring projects.

Clusia hilariana and Eugenia uniflora as bioindicators of atmospheric pollutants emitted by an iron pelletizing factory in Brazil.

The objectives of this work were to evaluate if the pollution emitted by the pelletizing factory causes visual symptoms and/or anatomical changes in exposed Eugenia uniflora and Clusia hilariana, in active biomonitoring, at different distances from a pelletizing factory. We characterize the symptomatology, anatomical, and histochemistry alterations induced in the two species. There was no difference in the symptomatology in relation to the different distances of the emitting source. The foliar symptoms found in C. hilariana were chlorosis, necrosis, and foliar abscission and, in E. uniflora, were observed necrosis punctuais, purple spots in the leaves, and increase in the emission of new leaves completely purplish. The two species presented formation of a cicatrization tissue. E. uniflora presented reduction in the thickness of leaf. In C. hilariana, it was visualized hyperplasia of the cells and the adaxial epidermis did not appear collapsed due to thick cuticle and cuticular flanges. Leaves of C. hilariana showed positive staining for iron, protein, starch, and phenolic compounds. E. uniflora showed positive staining for total phenolic compounds and starch. Micromorphologically, there was accumulation of particulate matter on the leaf surface, obstruction of the stomata, and scaling of the epicuticular wax in both species. It was concluded that the visual and anatomical symptoms were efficient in the diagnosis of the stress factor. C. hilariana and E. uniflora showed to be good bioindicators of the atmospheric pollutants emitted by the pelletizing factory.

Glyphosate-induced oxidative stress in Arabidopsis thaliana affecting peroxisomal metabolism and triggers activity in the oxidative phase of the pentose phosphate pathway (OxPPP) involved in NADPH generation.

Glyphosate is a broad-spectrum systemic herbicide used worldwide. In susceptible plants, glyphosate affects the shikimate pathway and reduces aromatic amino acid synthesis. Using Arabidopsis seedlings grown in the presence of 20µM glyphosate, we analyzed H2O2, ascorbate, glutathione (GSH) and protein oxidation content as well as antioxidant catalase, superoxide dismutase (SOD) and ascorbate-glutathione cycle enzyme activity. We also examined the principal NADPH-generating system components, including glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), NADP-malic enzyme (NADP-ME) and NADP-isocitrate dehydrogenase (NADP-ICDH). Glyphosate caused a drastic reduction in growth parameters and an increase in protein oxidation. The herbicide also resulted in an overall increase in GSH content, antioxidant enzyme activity (catalase and all enzymatic components of the ascorbate-glutathione cycle) in addition to the two oxidative phase enzymes, G6PDH and 6PGDH, in the pentose phosphate pathway involved in NADPH generation. In this study, we provide new evidence on the participation of G6PDH and 6PGDH in the response to oxidative stress induced by glyphosate in Arabidopsis, in which peroxisomal enzymes, such as catalase and glycolate oxidase, are positively affected. We suggest that the NADPH provided by the oxidative phase of the pentose phosphate pathway (OxPPP) should serve to maintain glutathione reductase (GR) activity, thus preserving and regenerating the intracellular GSH pool under glyphosate-induced stress. It is particularly remarkable that the 6PGDH activity was unaffected by pro-oxidant and nitrating molecules such as H202, nitric oxide or peroxynitrite.

Arsenic accumulation in Brassicaceae seedlings and its effects on growth and plant anatomy.

We wished to evaluate the effects of arsenic on the morphology and anatomy of Brassica oleracea, Raphanus sativus, Brassica juncea, Brassica oleracea var. capitata and Brassica oleracea var. italica. Seeds were subjected to concentrations 0µM, 250µM, 350µM and 450µM arsenic in the form of sodium arsenate (Na2HAsO4·7H2O) during 12 days. All species accumulated more arsenic in the roots than in the shoots, except for B. oleracea var. capitata. There was no difference of translocation factor between species and treatments. Growth decrease was observed in roots of B. oleracea and R. sativus, and in shoots of R. sativus and B. oleracea var. italica. All species presented anatomical alterations in the roots, such as: cell hypertrophy, protoplast retraction, cellular plasmolysis, and necrotic regions. B. juncea presented collapse and hypertrophy of cells from the leaf blade tissues. Quantitative anatomical analyses performed on the root and leaves of B. oleracea and B. juncea revealed that arsenic interfered on the root vascular cylinder diameter and on height of epidermal cells of the adaxial leaf surface of both species. We concluded that arsenic was absorbed from the culture medium and induced alterations both on root and shoot growth of the seedlings. Retention of arsenic within the root was responsible for major damage in this organ.

Differential responses of C3 and CAM native Brazilian plant species to a SO2- and SPMFe-contaminated Restinga.

Aiming to evaluate responses in terms of growth rates, physiological parameters, and degree of sensitivity to SO2 and SPMFe in Eugenia uniflora L. (Myrtaceae, a C3 species) and Clusia hilariana Schlecht (Clusiaceae, a CAM species); saplings were exposed to emissions from a pelletizing factory for 7 months. The species were distributed along a transect (200, 500, 800, 1400, and 1700 m away from the emission source), and analyses were performed after 71, 118, and 211 days of exposure to the pollutants. E. uniflora received higher superficial deposition of particulate iron. The highest total iron foliar contents were observed 200 m away from the emission source in both plant species, while the highest total sulfur foliar contents were observed 200 m away in C. hilariana and 800 m away in E. uniflora. E. uniflora presented decreased values of height growth rate, number of necrotic leaves, chlorophyll analysis (SPAD index) and transpiration, in relation to the distances from the emission source. C. hilariana showed decreased values of height growth rate, number of leaves, number of necrotic leaves, total ionic permeability, stomatal conductance, transpiration, net CO2 assimilation, and total dry matter, in relation to distances from the emission source. In relation to the days of exposure, both species presented increased number of necrotic leaves and foliar phytotoxicity index, and decreased values in the chlorophyll analysis. The two native plant species, both of which occur in the Brazilian Restinga, showed damage when exposed to emissions from an iron ore pelletizing factory. C. hilariana was considered the most sensitive species due to the decreased values in a higher number of variables after exposition.

Morphoanatomical responses induced by excess iron in roots of two tolerant grass species.

We aimed to verify whether morphoanatomic alterations occur in response to excess iron, in roots of Setaria parviflora and Paspallum urvillei (Poaceae), and to localize the presence of the sites of iron accumulation. Plants were subjected to 0.009, 1, 2, 4, and 7 mM Fe-EDTA in nutrient solution. Both species presented iron contents in the roots above the critical toxicity level. The presence of iron plaque on roots of the two species was confirmed, and it may have reduced iron absorption by the plants. Roots from the two species showed typical visual symptoms of stress by excess iron: change in color and mucilaginous and flaccid appearance. Anatomical damage was observed in both species: aerenchyma disruption, alterations in endodermal cells, and irregular shape of both vessel and sieve tube elements. The metal was histolocalized in the cortex and in protoxylem and metaxylem cell walls in both species, which suggests a detoxification strategy for the excess iron. Phenolic compounds were not histolocalized in roots. Microscopic analyses were therefore effective in evaluating the real damage caused by excess iron.

Protective effect of Mn(III)-desferrioxamine B upon oxidative stress caused by ozone and acid rain in the Brazilian soybean cultivar Glycine max "Sambaiba".

This study aimed to investigate the effects of the Mn complex (Mn(III)-desferrioxamine B (MnDFB)) on oxidative stress in the Brazilian soybean cultivar Glycine max "Sambaiba" following exposure to ozone and acid rain. We determined the suitable dose of MnDFB to apply to G. max seedlings using a dose-response curve. The highest superoxide dismutase (SOD) activity and Mn content in leaves were found upon the application of 8 µM MnDFB. Thus, G. max seedlings pretreated with 8 µM MnDFB were individually exposed to ozone and acid rain simulated. Pretreatment with MnDFB reduced lipid peroxidation upon ozone exposure and increased SOD activity in leaves; it did not alter the metal content in any part of the plant. Conversely, following acid rain exposure, neither the metal content in leaves nor SOD enzyme activity were directly affected by MnDFB, unlike pH. Our findings demonstrated that exogenous MnDFB application before ozone exposure may modulate the MnSOD, Cu/ZnSOD, and FeSOD activities to combat the ROS excess in the cell. Here, we demonstrated that the applied dose of MnDFB enhances antioxidative defenses in soybean following exposure to acid rain and especially to ozone.

Leaf morphoanatomy of species tolerant to excess iron and evaluation of their phytoextraction potential.

Setaria parviflora (Poir.) Kerguélen and Paspalum urvillei Steudel are grasses that grow naturally in a soil with high iron contents. This study aimed to characterize morphoanatomically and histochemically the iron phytotoxicity on leaves and evaluate the phytoextraction potential of these grasses. Saplings were cultivated in hydroponic solution with and without excess Fe-EDTA. Regarding measurements taken on leaves, reduction was observed among treatments of Fe-EDTA on height values of abaxial epidermis and bundle sheath in both species. As for iron histolocalization, stronger reaction was observed in leaves of S. parviflora, in comparison with P. urvillei. Anatomical damage, such as protoplast retraction, irregular xylem, changes in cell volume, and cell collapse, and visual symptoms, like leaf bronzing, chlorosis, and necrosis, were similar in both species when exposed to excess iron; however, P. urvillei showed more severe damage. This species accumulated more iron in shoots than S. parviflora and therefore is more favorable for use in phytoextraction. The root system of both species accumulated higher iron concentrations in relation to shoots.

Tolerance to iron accumulation and its effects on mineral composition and growth of two grass species.

This study aimed to assess the influence of excess iron on the capacity of accumulation of this heavy metal, mineral composition, and growth of Setaria parviflora and Paspalum urvillei. Seedlings were submitted to 0.009; 1; 2; 4; and 7 mM of Fe-EDTA. In both species there was an increase in the concentration of Fe, Zn, P, and Ca and a decrease in Mn, K, and Mg in the iron plaque. Both species accumulated more iron in roots. In the shoots, S. parviflora showed higher iron content, except at 7 mM. Iron altered the contents of Fe, Cu, K, and Mg in roots, and of Fe, Mn, Zn, N, P, K, Ca, and Mg in shoots. The two species tolerated high iron concentrations and accumulated high content of this element in both shoots and roots. The iron did not reduce their growth. Both species are indicated for studies aiming restoration of iron-contaminated areas.

Somatic embryogenesis of a wild passion fruit species Passiflora cincinnata Masters: histocytological and histochemical evidences.

The characterization of cellular changes that occur during somatic embryogenesis is essential for understanding the factors involved in the transition of somatic cells into embryogenically competent cells and determination of cells and/or tissues involved. The present study describes the anatomical and ultrastructural events that lead to the formation of somatic embryos in the model system of the wild passion fruit (Passiflora cincinnata). Mature zygotic embryos were inoculated in Murashige and Skoog induction media supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Zygotic embryo explants at different development stages were collected and processed by conventional methods for studies using light, scanning, and transmission electron microscopy (TEM). Histochemical tests were used to examine the mobilization of reserves. The differentiation of the somatic embryos began in the abaxial side of the cotyledon region. Protuberances were formed from the meristematic proliferation of the epidermal and mesophyll cells. These cells had large nuclei, dense cytoplasm with a predominance of mitochondria, and a few reserve compounds. The protuberances extended throughout the abaxial surface of the cotyledons. The ongoing differentiation of peripheral cells of these structures led to the formation of proembryogenic zones, which, in turn, dedifferentiated into somatic embryos of multicellular origin. In the initial stages of embryogenesis, the epidermal and mesophyll cells showed starch grains and less lipids and protein reserves than the starting explant. These results provide detailed information on anatomical and ultrastructural changes involved in the acquisition of embryogenic competence and embryo differentiation that has been lacking so far in Passiflora.

Iron plaque formation and morphoanatomy of roots from species of restinga subjected to excess iron.

The restingas, a sandy coastal plain ecosystem of Brazil, have received an additional amount of iron due to the activity of mining industries. The present study aims to characterize morphoanatomically and histochemically the iron plaque formation on roots of Ipomoea pes-caprae L. and Canavalia rosea DC, cultivated in hydroponic solution with and without excess iron. The iron plaque formation as well as changes in the external morphology of the lateral roots of both species were observed after the subjection to excess iron. Changes in the nutrient uptake, and in the organization and form of the pericycle and cortex cells were observed for both species. Scanning electron microscopy showed evident iron plaques on the whole surface of the root. The iron was histolocalized in all root tissues of both species. The species of restinga studied here formed iron plaque in their roots when exposed to excess of this element, which may compromise their development in environments polluted by particulated iron.