Effects on growth, antioxidant enzyme activity and levels of extracellular proteins in the green alga Chlorella vulgaris exposed to crude cyanobacterial extracts and pure microcystin and cylindrospermopsin

Abstract

Toxic cyanobacteria and cyanotoxins have been pointed as important players in the control of phytoplankton diversity and species abundance, causing ecological unbalances and contamination of the environment. In vitro experiments have been undertaken to address the impact of toxic cyanobacteria in green algae. In this regard the aim of this work was to compare the toxicity of two cyanobacteria species, Aphanizomenon ovalisporum and Microcystis aeruginosa, to the green alga Chlorella vulgaris by assessing culture growth when exposed for three and seven days to (I) cyanobacterial cell extracts and (II) pure toxins microcystin-LR (MC-LR) and cylindrospermopsin (CYN). The biochemical response of the green alga to pure toxins was also characterized, through the activity of the antioxidant markers glutathione S-transferase (GST) and glutathione peroxidase (GPx) and the expressed extracellular proteins in seven-day exposed cultures. A. ovalisporum crude extracts were toxic to C. vulgaris. Pure toxins up to 179.0µg/L, on the other hand, stimulated the green alga growth. Growth results suggest that the toxicity of A. ovalisporum extracts is likely due to a synergistic action of CYN and other metabolites produced by the cyanobacterium. Regarding the green alga antioxidant defense mechanism, CYN at 18.4 and 179.0µg/L increased the activity of GPx and GST while MC-LR inhibited the enzymes' activity at a concentration of 179.0µg/L demonstrating a contrasting mode of action. Moreover the identification of F-ATPase subunit, adenylate cyclase, sulfate ABC transporter, putative porin, aspartate aminotransferase, methylene-tetrahydrofolate dehydrogenase and chlorophyll a binding proteins in the culture medium of C. vulgaris indicates that biochemical processes involved in the transport of metabolites, photosynthesis and amino acid metabolism are affected by cyanobacterial toxins and may contribute to the regulation of green alga growth.