Aluminum promotes changes in rice root structure and ascorbate and glutathione metabolism.

In acidic soil, aluminum (Al) ionizes into trivalent cation and becomes highly toxic to plants. Thus, the objectives of this work were (i) to evaluate the Al concentration and identify sites of Al toxicity and its effect on the structure on rice root tips and (ii) to elucidate the adjustments involved in the activities/contents of enzymes/compounds in the roots against Al. For this, two genotypes with contrasting Al tolerance were used. Our results showed that the root length of the tolerant genotype was not affected after Al exposure. We also observed that both the genotypes used strategies to avoid Al uptake, such as the overlap of P and Al in the tolerant genotype and the presence of border cells in the sensitive genotype, which proved inefficient. In the tolerant genotype, other external Al detoxification mechanisms may have contributed to the lower Al concentration in roots and lower fluorescence of the Al-lumogallion complex. Additionally, both genotypes present the activation of key enzymes to decrease oxidative stress, such as CAT, POX, APX, and DHAR, and a more reducing redox environment, mainly due to the increase in the AA/DHA ratio. However, higher total ascorbate, AA, total glutathione, and GSH contents associated with higher SOD, GPX, and GR activities contributed to the reduction of oxidative stress in the tolerant genotype after Al exposure. Furthermore, there was a strong association between the sensitive genotype to Al concentration, O2 •- content, and MDA amount; therefore, these traits can be used as sensitivity indicators in Al studies.

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.

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.

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.

Separation of Plant 6-Phosphogluconate Dehydrogenase (6PGDH) Isoforms by Non-denaturing Gel Electrophoresis.

6-Phosphogluconate dehydrogenase (6PGDH; EC 1.1.1.44) catalyzes the third and irreversible reaction of the pentose phosphate pathway (PPP). It carries out the oxidative decarboxylation of the 6-phosphogluconate to yield ribulose-5-phosphate, carbon dioxide and NADPH. In higher plants, 6PGDH has several subcellular localizations including cytosol, chloroplast, mitochondria and peroxisomes ( Corpas et al., 1998 ; Krepinsky et al., 2001 ; Mateos et al., 2009 ; Fernández-Fernández and Corpas, 2016; Hölscher et al., 2016 ). Using Arabidopsis thaliana as plant model and sweet pepper (Capsicum annuum L.) fruits as a plant with agronomical interest, this protocol illustrates how to prepare the plant extracts for the separation of the potential 6PGDH isoforms by electrophoresis on 6% polyacrylamide non-denaturing gels. Thus, this method allows detecting three 6PGDH isoforms in Arabidopsis seedlings and two 6PGDH isoforms in sweet pepper fruits.

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.