Arsenic hyperaccumulation in Pityrogramma calomelanos L. (Link): adaptive traits to deal with high metalloid concentrations.

Pityrogramma calomelanos is interestingly the single non-Pteris arsenic (As)-hyperaccumulating fern. It has been pointed as a potential species for phytoremediation and a model plant to study the As toxicity and its mechanisms of action. In order to investigate the morphoanatomical traits associated to As tolerance, P. calomelanos plants were exposed to different As concentrations in hydroponic solution. At low As dose (1 mM As), 90% of the As accumulated in plants was allocated in shoots, and no symptoms of As stress were observed in fronds and roots. Under higher As exposure (10 and 30 mM As), 81-74% of the total As in plants was present in shoots, and apical and marginal necroses on pinnae were observed. Anatomical observations showed that As induces damages mainly in the secondary veins and adjacent cells. High amounts of phenols were observed in pinna tissues of control and treated plants. In the roots, As promoted slight alterations as detachment of border-like cells and accumulation of granular substances in cortical cells. The high root-to-shoot As translocation and the constitutive presence of phenols and border-like cells protecting the root tips showed to be adaptive traits that allow P. calomelanos to survive in contaminated sites.

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.

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.

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.

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.