Chlorophyll fluorescence induction curves were used as a means to assess the functional condition of the photosynthetic apparatus in cells of the halotolerant green microalga Dunaliella maritima (Massjuk) (division Chlorophyta) exposed to hyperosmotic salt shock of various intensities. The shock was caused by the transfer of algal cells grown in the medium with 0.5 M NaCl to the media with elevated NaCl concentra tions (1.0, 1.5, and 2.0 M). Parameters of chlorophyll fluorescence (F 0 , F m , F 0 ', ) were measured by means of a specialized pulse amplitude modulation fluorometer PAM 2100. In addition, the rate of photo synthetic oxygen evolution as well as the intracellular Na + and glycerol content (the main osmolyte in this microalga) were determined. The hyperosmotic salt shock was found to elevate the intracellular Na + content and reduce the functional activity of PSII in D. maritima. The suppression of PSII activity was evident from the decrease in the maximal quantum yield of photochemical energy conversion in PSII, the decreased rate of linear electron transport, the increased reduction of the primary acceptor Q A , and the suppression of pho tosynthetic O 2 evolution. The functional activity of PSII recovered gradually along with restoration of osmotic and ionic balance in algal cells. It is proposed that PSI ensures energy supply during cell responses of D. maritima to hyperosmotic salt shock.

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The green microalga genus Dunaliella is mostly comprised of species that exhibit a wide range of salinity tolerance, including inhabitants of hyperhaline reservoirs. Na+ content in Dunaliella cells inhabiting saline environments is maintained at a fairly low level, comparable to that in the cells of freshwater organisms. However, despite a long history of studying the physiological and molecular mechanisms that ensure the ability of halotolerant Dunaliella species to survive at high concentrations of NaCl, the question of how Dunaliella cells remove excess Na+ ions entering from the environment is still debatable. For thermodynamic reasons it should be a primary active mechanism; for example, via a Na+-transporting ATPase, but the molecular identification of Na+-transporting mechanism in Dunaliella has not yet been carried out. Formerly, in the euryhaline alga D. maritima, we functionally identified Na+-transporting P-type ATPase in experiments with plasma membrane (PM) vesicles which were isolated from this alga. Here we describe the cloning of two putative P-type ATPases from D. maritima, DmHA1 and DmHA2. Phylogenetic analysis showed that both ATPases belong to the clade of proton P-type ATPases, but the similarity between DmHA1 and DmHA2 is not high. The expression of DmHA1 and DmHA2 in D. maritima cells under hyperosmotic salt shock was studied by qRT-PCR. Expression of DmHA1 gene decreases and remains at a relatively low level during the response of D. maritima cells to hyperosmotic salt shock. In contrast, expression of DmHA2 increases under hyperosmotic salt shock. This indicates that DmHA2 is important for overcoming hyperosmotic salt stress by the algal cells and as an ATPase it is likely directly involved in transport of Na+ ions. We assume that it is the DmHA2 ATPase that represents the Na+-transporting ATPase.

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