A comparative study of the accumulation and detoxification of copper and zinc in Chlamydomonas reinhardtii: The role of extracellular polymeric substances.

Extracellular polymeric substances (EPS) form an interface between microalgae and the surrounding water environment. Copper (Cu) and zinc (Zn) are essential micronutrients but may negatively affect microbial growth when their concentrations reach toxic thresholds. However, how EPS affect the accumulation and resistance of Cu and Zn in microalgae remains largely unknown. Here, we investigated EPS production upon Cu/Zn exposure and compared the tolerance strategies to the two metals by Chlamydomonas reinhardtii with and without EPS. Microalgal EPS synthesis was induced by Cu/Zn treatments, and the functional groups of polysaccharides and proteins were involved in complexation with metal ions. The extraction of EPS aggravated the toxicity and reduced the removal of metals from solution, but the effect was more pronounced for Cu than for Zn. Copper bound on the cell surface accounted for 54.6 ± 2.0 % of the Cu accumulated by C. reinhardtii, whose EPS components strongly correlated with Cu adsorption. In contrast, 74.3 ± 3.0 % of accumulated Zn was absorbed in cells, and glutathione synthesis was significantly induced. Redundancy and linear correlation analyses showed that the polysaccharide, protein and DNA contents in EPS were significantly correlated with Cu accumulation, absorption and adsorption but not with Zn. Data fitted to a Michaelis-Menten model further showed that the EPS-intact cells had higher binding capacity for Cu2+ but not for Zn2+. These differential impacts of EPS on Cu/Zn sorption and detoxification contribute to a more comprehensive understanding of the roles of microalgal EPS in the biogeochemical cycle of metals.

Contrasting detoxification mechanisms of Chlamydomonas reinhardtii under Cd and Pb stress.

Chlamydomonas reinhardtii has been frequently investigated for its resistance to metals; however, few studies have systematically compared the intracellular and extracellular processes involved in the detoxification of Cd and Pb by this microalga. We found that C. reinhardtii was more tolerant to Pb (concentration for 50% of the maximal effect; EC50: 29.48 ± 8.83 mg L-1) than to Cd (EC50: 12.48 ± 1.30 mg L-1) after 96 h of exposure. Extracellular polymeric substances (EPS), intracellular starch granules, lipid droplets, and glutathione were significantly increased under Cd and Pb treatments. Lead-containing particles were formed outside of the cells exposed to 30 mg L-1 of Pb, whereas no minerals were present when Cd was added. Various EPS functional groups, including -COOH, C-O-C (polysaccharides), and amide I and II (proteins), were involved in the interactions with Cd and Pb. The Pb removal rate (60.46-78.27%) by C. reinhardtii was higher than that of Cd (50.61-59.38%), and the microalgal cells with intact EPS bound more metals than those without EPS. Adsorption accounted for 79.62% of the total Cd accumulation in the low-Cd treatment, whereas absorption dominated the Pb accumulation at low Pb concentrations. The distributions of Cd and Pb in and out of the microalgal cells were reversed when the concentrations of the two metals increased. The detoxification strategies of C. reinhardtii for Cd and Pb were completely different, and these findings may assist in the phycoremediation of metal pollution in aquatic environments.