Phenotypic Diversity and Plasticity of Photoresponse Across an Environmentally Contrasting Family of Phytoflagellates.

Organisms often employ ecophysiological strategies to exploit environmental conditions and ensure bio-energetic success. However, the many complexities involved in the differential expression and flexibility of these strategies are rarely fully understood. Therefore, for the first time, using a three-part cross-disciplinary laboratory experimental analysis, we investigated the diversity and plasticity of photoresponsive traits employed by one family of environmentally contrasting, ecologically important phytoflagellates. The results demonstrated an extensive inter-species phenotypic diversity of behavioural, physiological, and compositional photoresponse across the Chlamydomonadaceae, and a multifaceted intra-species phenotypic plasticity, involving a broad range of beneficial photoacclimation strategies, often attributable to environmental predisposition and phylogenetic differentiation. Deceptively diverse and sophisticated strong (population and individual cell) behavioural photoresponses were observed, with divergence from a general preference for low light (and flexibility) dictated by intra-familial differences in typical habitat (salinity and trophy) and phylogeny. Notably, contrasting lower, narrow, and flexible compared with higher, broad, and stable preferences were observed in freshwater vs. brackish and marine species. Complex diversity and plasticity in physiological and compositional photoresponses were also discovered. Metabolic characteristics (such as growth rates, respiratory costs and photosynthetic capacity, efficiency, compensation and saturation points) varied elaborately with species, typical habitat (often varying more in eutrophic species, such as Chlamydomonas reinhardtii), and culture irradiance (adjusting to optimise energy acquisition and suggesting some propensity for low light). Considerable variations in intracellular pigment and biochemical composition were also recorded. Photosynthetic and accessory pigments (such as chlorophyll a, xanthophyll-cycle components, chlorophyll a:b and chlorophyll a:carotenoid ratios, fatty acid content and saturation ratios) varied with phylogeny and typical habitat (to attune photosystem ratios in different trophic conditions and to optimise shade adaptation, photoprotection, and thylakoid architecture, particularly in freshwater environments), and changed with irradiance (as reaction and harvesting centres adjusted to modulate absorption and quantum yield). The complex, concomitant nature of the results also advocated an integrative approach in future investigations. Overall, these nuanced, diverse, and flexible photoresponsive traits will greatly contribute to the functional ecology of these organisms, addressing environmental heterogeneity and potentially shaping individual fitness, spatial and temporal distribution, prevalence, and ecosystem dynamics.

Nitrate or ammonium: Influences of nitrogen source on the physiology of a green alga.

In freshwaters, algal species are exposed to different inorganic nitrogen (Ni) sources whose incorporation varies in biochemical energy demand. We hypothesized that due to the lesser energy requirement of ammonium ( NH 4 + )-use, in contrast to nitrate ( NO 3 - )-use, more energy remains for other metabolic processes, especially under CO2- and phosphorus (Pi) limiting conditions. Therefore, we tested differences in cell characteristics of the green alga Chlamydomonas acidophila grown on NH 4 + or NO 3 - under covariation of CO2 and Pi-supply in order to determine limitations, in a full-factorial design. As expected, results revealed higher carbon fixation rates for NH 4 + -grown cells compared to growth with NO 3 - under low CO2 conditions. NO 3 - -grown cells accumulated more of the nine analyzed amino acids, especially under Pi-limited conditions, compared to cells provided with NH 4 + . This is probably due to a slower protein synthesis in cells provided with NO 3 - . In contrast to our expectations, compared to NH 4 + -grown cells NO 3 - -grown cells had higher photosynthetic efficiency under Pi-limitation. In conclusion, growth on the Ni-source NH 4 + did not result in a clearly enhanced Ci-assimilation, as it was highly dependent on Pi and CO2 conditions (replete or limited). Results are potentially connected to the fact that C. acidophila is able to use only CO2 as its inorganic carbon (Ci) source.

Decreased phosphorus incorporation explains the negative effect of high iron concentrations in the green microalga Chlamydomonas acidophila.

The green microalga Chlamydomonas acidophila is an important primary producer in very acidic lakes (pH 2.0-3.5), characterized by high concentrations of ferric iron (up to 1 g total Fe L-1) and low rates of primary production. It was previously suggested that these high iron concentrations result in high iron accumulation and inhibit photosynthesis in C. acidophila. To test this, the alga was grown in sterilized lake water and in medium with varying total iron concentrations under limiting and sufficient inorganic phosphorus (Pi) supply, because Pi is an important growth limiting nutrient in acidic waters. Photosynthesis and growth of C. acidophila as measured over 5 days were largely unaffected by high total iron concentrations and only decreased if free ionic Fe3+ concentrations exceeded 100 mg Fe3+ L-1. Although C. acidophila was relatively rich in iron (up to 5 mmol Fe: mol C), we found no evidence of iron toxicity. In contrast, a concentration of 260 mg total Fe L-1 (i.e. 15 mg free ionic Fe3+ L-1), which is common in many acidic lakes, reduced Pi-incorporation by 50% and will result in Pi-limited photosynthesis. The resulting Pi-limitation present at high iron and Pi concentrations was illustrated by elevated maximum Pi-uptake rates. No direct toxic effects of high iron were found, but unfavourable chemical Pi-speciation reduced growth of the acidophile alga.

Environmental concentrations of pharmaceuticals directly affect phytoplankton and effects propagate through trophic interactions.

Pharmaceuticals are found in freshwater ecosystems where even low concentrations in the range of ng L-1 may affect aquatic organisms. In the current study, we investigated the effects of chronic exposure to three pharmaceuticals on two microalgae, a potential modulation of the effects by additional inorganic phosphorus (Pi) limitation, and a potential propagation of the pharmaceuticals' effect across a trophic interaction. The latter considers that pharmaceuticals are bioaccumulated by algae, potentially metabolized into more (or less) toxic derivates and consequently consumed by zooplankton. We cultured Acutodesmus obliquus and Nannochloropsis limnetica in Pi-replete and Pi-limited medium contaminated with one of three commonly human used pharmaceuticals: fluoxetine, ibuprofen, and propranolol. Secondly, we tested to what extent first level consumers (Daphnia magna) were affected when fed with pharmaceutical-grown algae. Chronic exposure, covering 30 generations, led to (i) decreased cell numbers of A. obliquus in the presence of fluoxetine (under Pi-replete conditions) (ii) increased carotenoid to chlorophyll ratios in N. limnetica (under Pi-limited conditions), and (iii) increased photosynthetic yields in A. obliquus (in both Pi-conditions). In addition, ibuprofen affected both algae and their consumer: Feeding ibuprofen-contaminated algae to Pi-stressed D. magna improved their survival. We demonstrate, that even very low concentrations of pharmaceuticals present in freshwater ecosystems can significantly affect aquatic organisms when chronically exposed. Our study indicates that pharmaceutical effects can cross trophic levels and travel up the food chain.

Ecophysiological strategies for growth under varying light and organic carbon supply in two species of green microalgae differing in their motility.

Mixing events in stratified lakes result in microalgae being exposed to varying conditions in light and organic carbon concentrations. Stratified lakes consist of an upper illuminated strata and a lower, darker strata where organic carbon accumulates. Therefore, in this contribution we explore the importance of dissolved organic carbon for growth under various light intensities by measuring some ecophysiological adaptations in two green microalgae. We compared the non-motile Chlorella vulgaris with the flagellated Chlamydomonas acidophila under auto-, mixo-, and heterotrophic growth conditions. In both algae the maximum photosynthetic and growth rates were highest under mixotrophy, and both algae appeared inhibited in their phosphorus acquisition under heterotrophy. Heterotrophic conditions provoked the largest differences as C. vulgaris produced chlorophyll a in darkness and grew as well as in autotrophic conditions, whereas Chl. acidophila bleached and could not grow heterotrophically. Although the fatty acid composition of both phytoplankton species differed, both species reacted in a similar way to changes in their growth conditions, mainly by a decrease of C18:3n-3 and an increase of C18:1n-9 from auto- to heterotrophic conditions. The two contrasting responses within the group of green microalgae suggest that dissolved organic carbon has a high deterministic potential to explain the survival and behaviour of green algae in the deeper strata of lakes.

Ecophysiology matters: linking inorganic carbon acquisition to ecological preference in four species of microalgae (Chlorophyceae).

The effect of CO2 supply is likely to play an important role in algal ecology. Since inorganic carbon (Ci ) acquisition strategies are very diverse among microalgae and Ci availability varies greatly within and among habitats, we hypothesized that Ci acquisition depends on the pH of their preferred natural environment (adaptation) and that the efficiency of Ci uptake is affected by CO2 availability (acclimation). To test this, four species of green algae originating from different habitats were studied. The pH-drift and Ci uptake kinetic experiments were used to characterize Ci acquisition strategies and their ability to acclimate to high and low CO2 conditions and high and low pH was evaluated. Results from pH drift experiments revealed that the acidophile and acidotolerant Chlamydomonas species were mainly restricted to CO2 , whereas the two neutrophiles were efficient bicarbonate users. CO2 compensation points in low CO2 -acclimated cultures ranged between 0.6 and 1.4 µM CO2 and acclimation to different culture pH and CO2 conditions suggested that CO2 concentrating mechanisms were present in most species. High CO2 acclimated cultures adapted rapidly to low CO2 condition during pH-drifts. Ci uptake kinetics at different pH values showed that the affinity for Ci was largely influenced by external pH, being highest under conditions where CO2 dominated the Ci pool. In conclusion, Ci acquisition was highly variable among four species of green algae and linked to growth pH preference, suggesting that there is a connection between Ci acquisition and ecological distribution.

Photosynthetic sensitivity of phytoplankton to commonly used pharmaceuticals and its dependence on cellular phosphorus status.

Recently pharmaceuticals have become significant environmental pollutants in aquatic ecosystems, that could affect primary producers such as microalgae. Here we analyzed the effect of pharmaceuticals on the photosynthesis of microalgae commonly found in freshwater-two species of Chlorophyceae and a member of the Eustigmatophyceae, via PAM fluorometry. As pharmaceuticals, three medicines often consumed in households were chosen: (i) fluoxetine, an antidepressant, (ii) propranolol, a ß-blocker and (iii) ibuprofen, an anti-inflammatory and analgesic medicine. The EC50 for the quantum yield of photosystem II in phytoplankton acclimated to inorganic phosphorus (Pi)-replete and Pi-limited conditions was estimated. Acute toxicity experiments over a 5 h exposure revealed that Nannochloropsis limnetica was the least sensitive to pharmaceuticals in its photosynthetic yield out of all species tested. Although the estimation of sub-lethal effects can be vital in contrast to that of LC50s, the EC50 values in all species and for all medicines were orders of magnitude higher than concentrations found in polluted surface water. Chlamydomonas reinhardtii was the most sensitive to fluoxetine (EC50 of 1.6 mg L(-1)), and propranolol (EC50 of 3 mg L(-1)). Acutodesmus obliquus was most sensitive to ibuprofen (EC50 of 288 mg L(-1)). Additionally, the sensitivity to the pharmaceuticals changed under a Pi-limitation; the green algae became less sensitive to fluoxetine and propranolol. In contrast, Pi-limited algal species were more sensitive to ibuprofen. Our results suggest that the sensitivity of algae to pharmaceuticals is (i) highly compound- and species-specific and (ii) dependent on the cellular P status.

CO2 acquisition in Chlamydomonas acidophila is influenced mainly by CO2, not phosphorus, availability.

The extremophilic green microalga Chlamydomonas acidophila grows in very acidic waters (pH 2.3-3.4), where CO2 is the sole inorganic carbon source. Previous work has revealed that the species can accumulate inorganic carbon (Ci) and exhibits high affinity CO2 utilization under low-CO2 (air-equilibrium) conditions, similar to organisms with an active CO2 concentrating mechanism (CCM), whereas both processes are down-regulated under high CO2 (4.5 % CO2) conditions. Responses of this species to phosphorus (Pi)-limited conditions suggested a contrasting regulation of the CCM characteristics. Therefore, we measured external carbonic anhydrase (CAext) activities and protein expression (CAH1), the internal pH, Ci accumulation, and CO2-utilization in cells adapted to high or low CO2 under Pi-replete and Pi-limited conditions. Results reveal that C. acidophila expressed CAext activity and expressed a protein cross-reacting with CAH1 (the CAext from Chlamydomonas reinhardtii). Although the function of this CA remains unclear, CAext activity and high affinity CO2 utilization were the highest under low CO2 conditions. C. acidophila accumulated Ci and expressed the CAH1 protein under all conditions tested, and C. reinhardtii also contained substantial amounts of CAH1 protein under Pi-limitation. In conclusion, Ci utilization is optimized in C. acidophila under ecologically relevant conditions, which may enable optimal survival in its extreme Ci- and Pi-limited habitat. The exact physiological and biochemical acclimation remains to be further studied.

Elemental and fatty acid composition of snow algae in Arctic habitats.

Red, orange or green snow is the macroscopic phenomenon comprising different eukaryotic algae. Little is known about the ecology and nutrient regimes in these algal communities. Therefore, eight snow algal communities from five intensively tinted snow fields in western Spitsbergen were analysed for nutrient concentrations and fatty acid (FA) composition. To evaluate the importance of a shift from green to red forms on the FA-variability of the field samples, four snow algal strains were grown under nitrogen replete and moderate light (+N+ML) or N-limited and high light (-N+HL) conditions. All eight field algal communities were dominated by red and orange cysts. Dissolved nutrient concentration of the snow revealed a broad range of NH(+) (4) (<0.005-1.2 mg N l(-1)) and only low PO(3-) (4) (<18 µg P l(-1)) levels. The external nutrient concentration did not reflect cellular nutrient ratios as C:N and C:P ratios of the communities were highest at locations containing relatively high concentrations of NH(+) (4) and PO(3-) (4). Molar N:P ratios ranged from 11 to 21 and did not suggest clear limitation of a single nutrient. On a per carbon basis, we found a 6-fold difference in total FA content between the eight snow algal communities, ranging from 50 to 300 mg FA g C(-1). In multivariate analyses total FA content opposed the cellular N:C quota and a large part of the FA variability among field locations originated from the abundant FAs C18:1n-9, C18:2n-6, and C18:3n-3. Both field samples and snow algal strains grown under -N+HL conditions had high concentrations of C18:1n-9. FAs possibly accumulated due to the cessation of growth. Differences in color and nutritional composition between patches of snow algal communities within one snow field were not directly related to nutrient conditions. We propose that the highly patchy distribution of snow algae within and between snow fields may also result from differences in topographical and geological parameters such as slope, melting water rivulets, and rock formation.

Complementary ecophysiological strategies combine to facilitate survival in the hostile conditions of a deep chlorophyll maximum.

In the deep, cooler layers of clear, nutrient-poor, stratified water bodies, phytoplankton often accumulate to form a thin band or "deep chlorophyll maximum" (DCM) of ecological importance. Under such conditions, these photosynthetic microorganisms may be close to their physiological compensation points and to the boundaries of their ecological tolerance. To grow and survive any resulting energy limitation, DCM species are thought to exhibit highly specialised or flexible acclimation strategies. In this study, we investigated several of the adaptable ecophysiological strategies potentially employed by one such species, Chlamydomonas acidophila: a motile, unicellular, phytoplanktonic flagellate that often dominates the DCM in stratified, acidic lakes. Physiological and behavioural responses were measured in laboratory experiments and were subsequently related to field observations. Results showed moderate light compensation points for photosynthesis and growth at 22°C, relatively low maintenance costs, a behavioural preference for low to moderate light, and a decreased compensation point for photosynthesis at 8°C. Even though this flagellated alga exhibited a physiologically mediated diel vertical migration in the field, migrating upwards slightly during the day, the ambient light reaching the DCM was below compensation points, and so calculations of daily net photosynthetic gain showed that survival by purely autotrophic means was not possible. Results suggested that strategies such as low-light acclimation, small-scale directed movements towards light, a capacity for mixotrophic growth, acclimation to low temperature, in situ exposure to low O(2), high CO(2) and high P concentrations, and an avoidance of predation, could combine to help overcome this energetic dilemma and explain the occurrence of the DCM. Therefore, corroborating the deceptive ecophysiological complexity of this and similar organisms, only a suite of complementary strategies can facilitate the survival of C. acidophila in this DCM.

Independent colimitation for carbon dioxide and inorganic phosphorus.

Simultaneous limitation of plant growth by two or more nutrients is increasingly acknowledged as a common phenomenon in nature, but its cellular mechanisms are far from understood. We investigated the uptake kinetics of CO(2) and phosphorus of the algae Chlamydomonas acidophila in response to growth at limiting conditions of CO(2) and phosphorus. In addition, we fitted the data to four different Monod-type models: one assuming Liebigs Law of the minimum, one assuming that the affinity for the uptake of one nutrient is not influenced by the supply of the other (independent colimitation) and two where the uptake affinity for one nutrient depends on the supply of the other (dependent colimitation). In addition we asked whether the physiological response under colimitation differs from that under single nutrient limitation.We found no negative correlation between the affinities for uptake of the two nutrients, thereby rejecting a dependent colimitation. Kinetic data were supported by a better model fit assuming independent uptake of colimiting nutrients than when assuming Liebigs Law of the minimum or a dependent colimitation. Results show that cell nutrient homeostasis regulated nutrient acquisition which resulted in a trade-off in the maximum uptake rates of CO(2) and phosphorus, possibly driven by space limitation on the cell membrane for porters for the different nutrients. Hence, the response to colimitation deviated from that to a single nutrient limitation. In conclusion, responses to single nutrient limitation cannot be extrapolated to situations where multiple nutrients are limiting, which calls for colimitation experiments and models to properly predict growth responses to a changing natural environment. These deviations from single nutrient limitation response under colimiting conditions and independent colimitation may also hold for other nutrients in algae and in higher plants.

Interactions between P-limitation and different C conditions on the fatty acid composition of an extremophile microalga.

The extremophilic microalga Chlamydomonas acidophila inhabits very acidic waters (pH 2-3.5), where its growth is often limited by phosphorus (P) or colimited by P and inorganic carbon (CO(2)). Because this alga is a major food source for predators in acidic habitats, we studied its fatty acid content, which reflects their quality as food, grown under a combination of P-limited and different carbon conditions (either mixotrophically with light + glucose or at high or low CO(2), both without glucose). The fatty acid composition largely depended on the cellular P content: stringent P-limited cells had a higher total fatty acid concentration and had a lower percentage of polyunsaturated fatty acids. An additional limitation for CO(2) inhibited this decrease, especially reflected in enhanced concentrations of 18:3(9,12,15) and 16:4(3,7,10,13), resulting in cells relatively rich in polyunsaturated fatty acids under colimiting growth conditions. The percentage of polyunsaturated to total fatty acid content was positively related with maximum photosynthesis under all conditions applied. The two factors, P and CO(2), thus interact in their effect on the fatty acid composition in C. acidophila, and colimited cells P-limited algae can be considered a superior food source for herbivores because of the high total fatty acid content and relative richness in polyunsaturated fatty acids.

The expression of a carbon concentrating mechanism in Chlamydomonas acidophila under variable phosphorus, iron, and CO2 concentrations.

The CO(2) acquisition was analyzed in Chlamydomonas acidophila at pH 2.4 in a range of medium P and Fe concentrations and at high and low CO(2) condition. The inorganic carbon concentrating factor (CCF) was related to cellular P quota (Q(p)), maximum CO(2)-uptake rate by photosynthesis (V(max,O2)), half saturation constant for CO(2) uptake (K(0.5)), and medium Fe concentration. There was no effect of the medium Fe concentration on the CCF. The CCF increased with increasing Q(p) in both high and low CO(2) grown algae, but maximum Q(p) was 6-fold higher in the low CO(2) cells. In high CO(2) conditions, the CCF was low, ranging between 0.8 and 3.5. High CCF values up to 9.1 were only observed in CO(2)-limited cells, but P- and CO(2)-colimited cells had a low CCF. High CCF did not relate with a low K(0.5) as all CO(2)-limited cells had a low K(0.5) (<4 µM CO(2)). High C(i)-pools in cells with high Q(p) suggested the presence of an active CO(2)-uptake mechanism. The CCF also increased with increasing V(max,O2) which reflect an adaptation to the nutrient in highest demand (CO(2)) under balanced growth conditions. It is proposed that the size of the CCF in C. acidophila is more strongly related to porter density for CO(2) uptake (reflected in V(max,O2)) and less- to high-affinity CO(2) uptake (low K(0.5)) at balanced growth. In addition, high CCF can only be realized with high Q(p).

What physiological acclimation supports increased growth at high CO2 conditions?

Chlamydomonas acidophila Negoro is a green algal species abundant in acidic waters (pH 2-3.5), in which inorganic carbon is present only as CO(2). Previous studies have shown that aeration with CO(2) increased its maximum growth rate, suggesting CO(2) limitation under natural conditions. To unravel the underlying physiological mechanisms at high CO(2) conditions that enables increased growth, several physiological characteristics from high- and low-CO(2)-acclimated cells were studied: maximum quantum yield, photosynthetic O(2) evolution (P(max)), affinity constant for CO(2) by photosynthesis (K(0.5,p)), a CO(2)-concentrating mechanism (CCM), cellular Rubisco content and the affinity constant of Rubisco for CO(2) (K(0.5,r)). The results show that at high CO(2) concentrations, C. acidophila had a higher K(0.5,p), P(max), maximum quantum yield, switched off its CCM and had a lower Rubisco content than at low CO(2) conditions. In contrast, the K(0.5,r) was comparable under high and low CO(2) conditions. It is calculated that the higher P(max) can already explain the increased growth rate in a high CO(2) environment. From an ecophysiological point of view, the increased maximum growth rate at high CO(2) will likely not be realised in the field because of other population regulating factors and should be seen as an acclimation to CO(2) and not as proof for a CO(2) limitation.

Phosphorus acquisition by Chlamydomonas acidophila under autotrophic and osmo-mixotrophic growth conditions.

Chlamydomonas acidophila Negoro is a green algal species abundant in acidic waters where inorganic phosphorus (P(i)) and carbon (CO(2)) are considered the most important growth-limiting nutrients for the phytoplankton. This paper describes the P(i) uptake and growth kinetics under varying carbon supply by cultivating the alga autotrophically, with and without CO(2) aeration, and osmo-mixotrophically with glucose under low P(i) conditions at pH 2.7. The low minimum cellular phosphorus quota (Q(0); ranging from 0.6 to 1.1 mmol P mol(-1) C) suggested P(i)-limiting conditions under all different modes of carbon supply, and was lowest under CO(2)-aerated conditions. The threshold P(i) concentration for growth did not vary from zero, suggesting no detectable metabolic costs. Maximum P(i)-uptake rates (V(max)) were a better indication of P(i) limitation when compared with the affinity constant for P(i) uptake (K(m)), as V(max) was only high under P(i)-limited conditions whereas K(m) was low under both P(i)-limited and P(i)-replete conditions. Osmo-mixotrophic growth conditions did not result in decreased extracellular phosphatase activity, but often resulted in physiological characteristics comparable with CO(2)-aerated cells, suggesting intracellular CO(2) production by glucose respiration. In addition, at low CO(2) and in autotrophic conditions, C. acidophila had a higher Q(0), lower dissolved organic carbon concentration, lower maximum P(i)-uptake rates, and lower phosphatase activity, suggesting that growth was co-limited by CO(2) and P(i). Furthermore, cells may respond physiologically to both nutrient limitations simultaneously.

Is there really insufficient support for Tilman's R* concept? A comment on Miller et al.

Miller et al. (2005), in the American Naturalist (165:439-448), critically reviewed the applicability of Tilman's resource-ratio hypothesis. One of their conclusions was that only eight experimental papers support the R* concept, while five do not. We are familiar with some of the latter studies, and we question this conclusion. Our evaluation shows that 12 of the 13 articles investigated by Miller et al. support R* prediction, while one article does not fit the experimental conditions for a proper test. Moreover, the microbial and aquatic literature contains many more competition experiments consistent with the R* prediction. We therefore conclude that there is strong experimental support for the R* concept, at least from studies with bacteria, phytoplankton, and zooplankton.

Stress responses and metal tolerance of Chlamydomonas acidophila in metal-enriched lake water and artificial medium.

Chlamydomonas acidophila faces high heavy-metal concentrations in acidic mining lakes, where it is a dominant phytoplankton species. To investigate the importance of metals to C. acidophila in these lakes, we examined the response of growth, photosynthesis, cell structure, heat-shock protein (Hsp) accumulation, and metal adsorption after incubation in metal-rich lake water and artificial growth medium enriched with metals (Fe, Zn). Incubation in both metal-rich lake water and medium caused large decreases in photosystem II function (though no differences among lakes), but no decrease in growth rate (except for medium + Fe). Concentrations of small Hsps were higher in algae incubated in metal-rich lake-water than in metal-enriched medium, whereas Hsp60 and Hsp70A were either less or equally expressed. Cellular Zn and Fe contents were lower, and metals adsorbed to the cell surface were higher, in lake-water-incubated algae than in medium-grown cells. The results indicate that high Zn or Fe levels are likely not the main or only contributor to the low primary production in mining lakes, and multiple adaptations of C. acidophila (e.g., high Hsp levels, decreased metal accumulation) increase its tolerance to metals and permit survival under such adverse environmental conditions. Supposedly, the main stress factor present in the lake water is an interaction between low P and high Fe concentrations.

Temperature- and pH-dependent accumulation of heat-shock proteins in the acidophilic green alga Chlamydomonas acidophila.

Chlamydomonas acidophila, a unicellular green alga, is a dominant phytoplankton species in acidic water bodies, facing severe environmental conditions such as low pH and high heavy metal concentrations. We examined the pH-, and temperature-dependent accumulation of heat-shock proteins in this alga to determine whether heat-shock proteins play a role in adaptation to their environment. Our results show increased heat-shock proteins accumulation at suboptimal pHs, which were not connected with any change in intracellular pH. In comparison to the mesophilic Chlamydomonas reinhardtii, the acidophilic species exhibited significantly higher accumulations of heat-shock proteins under control conditions, indicating an environmental adaptation of increased basal levels of heat-shock proteins. The results suggest that heat-shock proteins might play a role in the adaptation of C. acidophila, and possibly other acidophilic algae, to their extreme environment.

The role of ultraviolet-adaptation of a marine diatom in photoenhanced toxicity of acridine.

Cultures of the marine diatom Phaeodactylum tricornutum were grown under laboratory light with a different fraction of ultraviolet radiation (UV) to study the potential role of photoadaptation in determining the sensitivity to photoenhanced toxicity of acridine. In short-term experiments, a higher acridine concentration was needed to inhibit the photosynthetic electron flux, monitored with chlorophyll a fluorescence, in algae exposed to fluorescent light (low UV) than to mercury light (high UV), consistent with the expected role of UV. The two types of light in long-term exposures led to changes in the pigment composition and photosystem I (PS I) to photosystem II (PS II) stoichiometry to optimize the utilization of fluorescent and mercury light. Despite the adaptation of the photosynthetic apparatus to a small fraction of UV, long-term exposure to mercury light did show a constant sensitivity of the photosynthetic efficiency of P. tricornutum to the phototoxic acridine. It is concluded that the prime receptor of photoenhanced toxicity may be unrelated to the photosynthetic machinery.