Biochemical characterization of peroxidases from the moss Dicranum scoparium

Abstract

Mosses are a convenient model to study stress responses of plants because of their remarkable stress tolerance. Peroxidase (EC 1.11.1.7) activities were tested in three moss species, namely Dicranum scoparium, Hylocomium splendens and Pleurozium schreberi growing together in the same location in a boreal forest. Peroxidase activity in D. scoparium was twice as high as in other mosses. Total peroxidase activity in unstressed D. scoparium was constitutively high; furthermore, long-term desiccation caused a significant increase in activity after 48 h of drying. Interestingly, when thalli desiccated for a week were rapidly rehydrated, peroxidase activity initially declined and then increased after 2 h rehydration. Diverse anionic and cationic isoforms were detected by native isoelectric focusing and PAGE of both crude extracts and partially purified peroxidases. The ability of peroxidases from D. scoparium to produce superoxide radical (O2•−) was confirmed using the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay and in-gel nitroblue tetrazolium chloride (NBT) staining; specific O2•− producing isoforms were revealed using 2D electrophoresis. Given a quinone and chelated Fe3+D. scoparium could produce extracellular hydroxyl radical (•OH), and production was increased by desiccation/rehydration stress. The possible roles of peroxidases and quinone reductases in apoplastic •OH production is discussed. Our data demonstrate that D. scoparium possesses high constitutive peroxidase activity that can be further increased by desiccation stress. Among the diverse moss peroxidases, some anionic isoforms displayed both pro- and antioxidative activities. These findings suggest that the ability of peroxidases to produce and detoxify reactive oxygen species is an evolutionarily ancient characteristic, important for plant stress tolerance.