Arsenic hyperaccumulation in Pityrogramma calomelanos (L.) Link (Pteridaceae): morphophysiological mechanisms of tolerance

• Ano de defesa: 2014
• Autor(a) principal: Campos, Naiara Viana
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: eng
• Instituição de defesa: Universidade Federal de Viçosa
• Palavras-chave em Português: Samambaias; Pityrogramma calomelanos; Pityrogramma calomelanos - Tolerância - Arsênio; Fitorremediação; Botânica
• Link de acesso: http://www.locus.ufv.br/handle/123456789/7564

Arsenic (As) is a metalloid ubiquitously distributed in the environment. The presence of As in drinking water is a serious health problem in many countries due to its genotoxic and mutagenic effects. The cleanup of contaminated water and soils requires the use of appropriate remediation techniques to prevent possible impacts on the ecosystems. As- hyperaccumulating ferns provide the possibility of developing cost-effective, and ecofriendly As phytoremediation programs. Pteris vittata L. was the first As hyperaccumulating fern described, and has been widely studied. Pityrogramma calomelanos L. (Link) is a less known hyperaccumulating fern that grows naturally in As-contaminated sites. This study aimed to characterize the morphoanatomical and physiological mechanisms involved in As accumulation and tolerance in P. calomelanos. Three experiments (E1, E2 and E3) were performed using ferns at different stages of development and exposed to As (E1: 1 mM As; E2-E3: 1, 10, and 30 mM As) for two (E2) or three (E1 and E3) weeks. Arsenic was supplied as a solution of sodium arsenate and was added to Hoagland’s nutrient solution (1/2 strenght). Plants grown in solution without As were used as control. The chlorophyll (chl) a fluorescence was evaluated by Imaging-PAM fluorometer (E2) and Mini-PAM portable fluorometer (E3; only dark parameters), and the chl content was estimated by portable chlorophyll meter (SPAD) in E1. At the end of the experiments, plants were sampled for physiological (E1 and E3), microscopic (E2), chemical (E1-E3) and micro-energy dispersive X-ray fluorescence spectrometry (μ-EDXRF) (E1 and E3) analyses. Pityrogramma calomelanos showed a high potential to As uptake, translocation and accumulation. Arsenic concentrations in fronds ranged from 3000 to 12000 mg kg-1 dry weight (DW) and was higher in E3. Plants exposed to 1 mM As (E1), ferns accumulated As mainly in pinnae (75 % as arsenite), whereas roots and stipes showed similar As contents (95 % and 74 % as arsenate, respectively). Arsenic partition was similar between E1 and E2, whereas in E3, stipes and roots showed As concentrations higher than those found in pinnae at 10 and 30 mM As in solution. Arsenic reduced the P concentrations in ferns of E1, but not in those from E2 and E3. In E1, As reduced the content of K, Fe and Mg, and the translocation of K and S. Arsenic did not change chl content in plants from E1 and E3, however, differences were observed in chl a fluorescence of plants exposed to 10 and 30 mM (E2). Ferns showed reduction in DW of pinnae at 30 mM As (E2), and apical and marginal necrosis in old fronds at 10 and 30 mM As (E2 and E3). In an anatomic view, these ferns (E2: 10-30 mM As) showed increase in phenols (healthy pinna areas) and tissue disruption mainly in the pinnule margins and near the secondary veins (necrotic pinna areas). Arsenic induced slight alterations in the radicular system (E2), such as progressive darkening of the root, detachment of border-like cells, increase in cell wall thickness and accumulation of granular compounds in cortical cells at 30 mM As. The μ-EDXRF analysis confirmed that As was preferentially located in apical and marginal regions of pinnules and around the veins. Pityrogramma calomelanos showed an efficient antioxidant defense system, as observed by the increase in the activities of antioxidant enzymes (E1 and E3) and increase in non-protein thiols (E1), phenols (E2 and E3), and other scavenging molecules (E3), especially in pinnae. At extremely high As concentrations (e.g. 30 mM As) these protective mechanisms became inefficient, increasing the concentration of oxygen reactive species and culminating in lipid peroxidation in fronds (E3). Roots exposed to 1 mM As also showed increase in lipid peroxidation. In E3, As accumulation promoted reduction or maintenance of metabolites associated with central carbon metabolism (e.g. glucose) and increased the total concentration of amino acids and proteins. Amino acids up-regulated by As were associated with S metabolism, photorespiration, osmoprotection and secondary metabolite biosynthesis. Altogether, these findings indicate that P. calomelanos can be successfully used for remediation of moderate As-contaminated sites. Adult plants with higher biomass (e.g. 5-7 fronds - E3) are preferable due to their higher potential to extract As from the medium. Experiments with low-dose and long-time exposures (chronic exposure) are needed to clarify the dose-response-mediated mechanisms of As-tolerance in P. calomelanos, in order to improve its performance in the field. The harvesting of plants as soon as the appearing of visual symptoms, such as necrosis and shriveling of young fronds, can also improve the phytoremediation efficiency.