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Two Chlamydomonas reinhardtii CW15 populations modified by an excess of copper in growth medium were obtained: a "Cu" population that was continuously grown under the selection pressure of 5 µM Cu 2+ (for at least 48 weeks) and the "Re" population, where a relatively short (9 week) exposure to elevated copper, necessary for acquiring tolerance, was followed by a prolonged period (at least 39 weeks) of cultivation at a normal (0.25 µM) copper concentration.Cells of the Cu population were able to multiply at a Cu 2+ concentration 16 times higher than that of the control population at a normal light intensity and at a Cu 2+ concentration 64 times higher when cultivated in dim light. The potential quantum yield of photosystem II (F V /F M ratio) under copper stress was also significantly higher for the Cu population than for Re and control populations.The Re population showed only residual tolerance towards the elevated concentration of copper, which is revealed by an F V /F M ratio slightly higher than in the control population under Cu 2+ stress in dim light or in darkness. We postulate that in the Chlamydomonas populations studied in this paper, at least two mechanisms of copper tolerance operate. The first mechanism is maintained during cultivation at a standard copper concentration and seems to be connected with photosynthetic apparatus. This mechanism, however, has only low adaptive value under excess of copper. The other mechanism, with a much higher adaptive value, is probably connected with Cu 2+ homeostasis at the cellular level, but is lost during cultivation at a normal copper concentration.

Copper is an essential micronutrient, but at supraoptimal concentrations it is also highly toxic, inducing oxidative stress and disrupting photosynthesis. The aim of the present study was to analyze selected protective mechanisms in strains of Chlamydomonas reinhardtii adapted and not adapted for growth in the presence of elevated copper concentrations. Two algal lines (tolerant and non-tolerant to high Cu2+ concentrations) were used in experiments to study photosynthetic pigment content, peroxidase activity, and non-photochemical quenching. The content of prenyllipids was studied in four different algal lines (two of the same as above and two new ones). The copper-adapted strains contained about 2.6 times more α-tocopherol and plastoquinol and about 1.7 times more total plastoquinone than non-tolerant strains. Exposure to excess copper led to oxidation of the plastoquinone pool in non-tolerant strains, whereas this effect was less pronounced or did not occur in copper-tolerant strains. Peroxidase activity was approximately 1.75 times higher in the tolerant strain than in the non-tolerant one. The increase in peroxidase activity in the tolerant strain was less pronounced when the algae were grown in dim light. In the tolerant line nonphotochemical quenching was induced faster and was usually about 20–30% more efficient than in the non-tolerant line. The improvement of antioxidant defense and photoprotection may be important factors in the evolutionary processes leading to tolerance to heavy metals.

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