TY - JOUR A1 - Dauvillee, David A1 - Chochois, Vincent A1 - Steup, Martin A1 - Haebel, Sophie A1 - Eckermann, Nora A1 - Ritte, Gerhard A1 - Ral, Jean-Philippe A1 - Colleoni, Christophe A1 - Hicks, Glenn A1 - Wattebled, Fabrice A1 - Deschamps, Philippe A1 - Lienard, Luc A1 - Cournac, Laurent A1 - Putaux, Jean-Luc A1 - Dupeyre, Danielle A1 - Ball, Steven G. T1 - Plastidial phosphorylase is required for normal starch synthesis in Chlamydomonas reinhardtii JF - The plant journal N2 - Among the three distinct starch phosphorylase activities detected in Chlamydomonas reinhardtii, two distinct plastidial enzymes (PhoA and PhoB) are documented while a single extraplastidial form (PhoC) displays a higher affinity for glycogen as in vascular plants. The two plastidial phosphorylases are shown to function as homodimers containing two 91-kDa (PhoA) subunits and two 110-kDa (PhoB) subunits. Both lack the typical 80-amino-acid insertion found in the higher plant plastidial forms. PhoB is exquisitely sensitive to inhibition by ADP-glucose and has a low affinity for malto-oligosaccharides. PhoA is more similar to the higher plant plastidial phosphorylases: it is moderately sensitive to ADP-glucose inhibition and has a high affinity for unbranched malto-oligosaccharides. Molecular analysis establishes that STA4 encodes PhoB. Chlamydomonas reinhardtii strains carrying mutations at the STA4 locus display a significant decrease in amounts of starch during storage that correlates with the accumulation of abnormally shaped granules containing a modified amylopectin structure and a high amylose content. The wild-type phenotype could be rescued by reintroduction of the cloned wild-type genomic DNA, thereby demonstrating the involvement of phosphorylase in storage starch synthesis. KW - Chlamydomonas KW - starch KW - amylopectin KW - (glycogen) starch phosphorylase Y1 - 2006 U6 - https://doi.org/10.1111/j.1365-313X.2006.02870.x SN - 0960-7412 VL - 48 IS - 2 SP - 274 EP - 285 PB - Blackwell CY - Oxford ER - TY - JOUR A1 - Winck, Flavia Vischi A1 - Kwasniewski, Miroslaw A1 - Wienkoop, Stefanie A1 - Müller-Röber, Bernd T1 - An optimized method for the isolation of nuclei from chlamydomas Reinhardtii (Chlorophyceae) JF - Journal of phycology N2 - The cell nucleus harbors a large number of proteins involved in transcription, RNA processing, chromatin remodeling, nuclear signaling, and ribosome assembly. The nuclear genome of the model alga Chlamydomonas reinhardtii P. A. Dang. was recently sequenced, and many genes encoding nuclear proteins, including transcription factors and transcription regulators, have been identified through computational discovery tools. However, elucidating the specific biological roles of nuclear proteins will require support from biochemical and proteomics data. Cellular preparations with enriched nuclei are important to assist in such analyses. Here, we describe a simple protocol for the isolation of nuclei from Chlamydomonas, based on a commercially available kit. The modifications done in the original protocol mainly include alterations of the differential centrifugation parameters and detergent-based cell lysis. The nuclei-enriched fractions obtained with the optimized protocol show low contamination with mitochondrial and plastid proteins. The protocol can be concluded within only 3 h, and the proteins extracted can be used for gel-based and non-gel-based proteomic approaches. KW - 2D gel electrophoresis KW - algae KW - Chlamydomonas KW - nuclear proteins KW - nucleus KW - proteomics Y1 - 2011 U6 - https://doi.org/10.1111/j.1529-8817.2011.00967.x SN - 0022-3646 VL - 47 IS - 2 SP - 333 EP - 340 PB - Wiley-Blackwell CY - Malden ER - TY - JOUR A1 - Piepho, Maike A1 - Arts, Michael T. A1 - Wacker, Alexander T1 - Species-specific variation in fatty acid concentrations of four phytoplankton species does phosphorus supply influence the effect of light intensity of temperature? JF - Journal of phycology N2 - We tested, in the laboratory, the influence of light intensity, temperature, and phosphorus (P) supply on fatty acid (FA) concentrations of four freshwater algae: the green algae Scenedesmus quadricauda (Turpin) Breb. and Chlamydomonas globosa J. Snow, the cryptophyte Cryptomonas ovata Ehrenb., and the diatom Cyclotella meneghiniana Kutz. We investigated the main and interactive effects of two variables on algal FA concentrations (i.e., light intensity and P supply or temperature and P supply). Interactive effects of light intensity and P supply were most pronounced in C. meneghiniana, but were also found in S. quadricauda and C. ovata. Changes in several saturated and unsaturated FA concentrations with light were more distinct in the low-P treatments than in the high-P treatments. Interactive effects of temperature and P supply on various FA concentrations were observed in all four species, but there was no consistent pattern. In lake ecosystems, P limitation often coincides with high light intensities and temperatures in summer. Therefore, it is important to examine how combinations of these environmental conditions affect FA concentrations of primary producers that are important sources of FAs for higher trophic levels. KW - Chlamydomonas KW - Cryptomonas KW - Cyclotella KW - fatty acids KW - light KW - lipids KW - phosphate KW - PUFA KW - Scenedesmus KW - temperature Y1 - 2012 U6 - https://doi.org/10.1111/j.1529-8817.2011.01103.x SN - 0022-3646 VL - 48 IS - 1 SP - 64 EP - 73 PB - Wiley-Blackwell CY - Malden ER - TY - JOUR A1 - Lachmann, Sabrina C. A1 - Maberly, Stephen C. A1 - Spijkerman, Elly T1 - ECOPHYSIOLOGY MATTERS: LINKING INORGANIC CARBON ACQUISITION TO ECOLOGICAL PREFERENCE IN FOUR SPECIES OF MICROALGAE (CHLOROPHYCEAE) JF - Journal of phycology N2 - The effect of CO2 supply is likely to play an important role in algal ecology. Since inorganic carbon (C-i) acquisition strategies are very diverse among microalgae and C-i availability varies greatly within and among habitats, we hypothesized that C-i acquisition depends on the pH of their preferred natural environment (adaptation) and that the efficiency of C-i 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 C-i uptake kinetic experiments were used to characterize C-i 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 mu M 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. C-i uptake kinetics at different pH values showed that the affinity for C-i was largely influenced by external pH, being highest under conditions where CO2 dominated the C-i pool. In conclusion, C-i acquisition was highly variable among four species of green algae and linked to growth pH preference, suggesting that there is a connection between C-i acquisition and ecological distribution. KW - acidophile KW - carbon acquisition KW - CCM KW - Chlamydomonas KW - Chlorella KW - CO2 supply KW - extremophile KW - inorganic carbon uptake kinetics KW - pH-drift KW - Scenedesmus Y1 - 2016 U6 - https://doi.org/10.1111/jpy.12462 SN - 0022-3646 SN - 1529-8817 VL - 52 SP - 1051 EP - 1063 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Spijkerman, Elly A1 - Stojkovic, Slobodanka A1 - Holland, Daryl A1 - Lachmann, Sabrina C. A1 - Beardall, John T1 - Nutrient induced fluorescence transients (NIFTs) provide a rapid measure of P and C (co-)limitation in a green alga JF - European journal of phycology N2 - Nutrient Induced Fluorescence Transients (NIFTs) have been shown to be a possible way of testing for the limiting nutrient in algal populations. In this study we tested the hypothesis that NIFTs can be used to detect a (co-)limitation for inorganic phosphorus (Pi) and CO2 in the green alga Chlamydomonas acidophila and that the magnitude of the NIFTs can be related to cellular P:C ratios. We show a co-limitation response for Pi and CO2 via traditional nutrient enrichment experiments in natural phytoplankton populations dominated by C. acidophila. We measured NIFT responses after a Pi- or a CO2-spike in C. acidophila batch cultures at various stages of Pi and inorganic C limitation. Significant NIFTs were observed in response to spikes in both nutrients. The NIFT response to a Pi-spike showed a strong negative correlation with cellular P:C ratio that was pronounced below 3 mmol P: mol C (equivalent to 0.2 pg P cell(-1)). Both cellular P and C content influenced the extent of the Pi-NIFT response. The NIFT response to a CO2-spike correlated to low CO2 culturing conditions and also had a negative correlation with cellular P content. A secondary response within the Pi-NIFT response was related to the CO2 concentration and potentially reflected co-limitation. In conclusion, NIFTs provided a quick and reliable method to detect the growth-limiting nutrient in an extremophile green alga, under Pi-, CO2- and Pi/CO2 (co-)limited growth conditions. KW - acidophile KW - Chlamydomonas KW - CO2 concentrating mechanism KW - CO2 limitation KW - extremophile KW - nutrient limitation KW - photosynthesis response KW - phytoplankton KW - stoichiometry Y1 - 2016 U6 - https://doi.org/10.1080/09670262.2015.1095355 SN - 0967-0262 SN - 1469-4433 VL - 51 SP - 47 EP - 58 PB - Hindawi CY - Abingdon ER - TY - JOUR A1 - Lachmann, Sabrina C. A1 - Maberly, Stephen C. A1 - Spijkerman, Elly T1 - Species-specific influence of P-i-status on inorganic carbon acquisition in microalgae (Chlorophyceae) JF - Botany N2 - Inorganic phosphorus (P-i) is often the primary limiting nutrient in freshwater ecosystems. Since P(i-)limitation affects energy transduction, and inorganic carbon (C-i) acquisition can be energy demanding, C(i-)acquisition strategies were compared in four species of green algae grown under P-i-replete and P-i-limited conditions predominantly at low and partly at high CO2. Although P-i-limitation was evident by the 10-fold higher cellular C:P ratio and enhanced phosphatase activity, it only decreased C-i-acquisition to a small extent. Nonetheless, the effects of Pi-limitation on both CO2 and HCO3- acquisition were demonstrated. Decreased CO2 acquisition under conditions of Pi limitation was mainly visible in the maximum uptake rate (V-max) and, for the neutrophile Scenedesmus vacuolatus, in the affinity for CO2 acquisition. Discrimination against C-13 was higher under P-i-limited, high CO2 conditions, compared with P-i-replete, highCO(2) conditions, in Chlamydomonas acidophila and S. vacuolatus. In the pH-drift experiments, HCO3- acquisition was reduced in P-i-limited C. reinhardtii. In general, energy demanding bicarbonate uptake was indicated by the less strong discrimination against (13)Cunder lowCO(2) conditions in the neutrophiles (HCO3- users), separating them from the acidophilic or acidotolerant species (CO2 users). The high variability of the influence of Pi supply among different green algal species is linked to their species-specific C(i-)acquisition strategies. KW - CCM KW - Chlamydomonas KW - inorganic carbon uptake kinetics KW - inorganic phosphorus limitation KW - stable carbon isotope discrimination Y1 - 2017 U6 - https://doi.org/10.1139/cjb-2017-0082 SN - 1916-2790 SN - 1916-2804 VL - 95 SP - 943 EP - 952 PB - NRC Research Press CY - Ottawa ER - TY - JOUR A1 - Spijkerman, Elly A1 - Behrend, Hella A1 - Fach, Bettina A1 - Gaedke, Ursula T1 - Decreased phosphorus incorporation explains the negative effect of high iron concentrations in the green microalga Chlamydomonas acidophila JF - The science of the total environment : an international journal for scientific research into the environment and its relationship with man N2 - 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. KW - Chlamydomonas KW - Ecotoxicology KW - Extreme environment KW - Iron toxicity KW - Phosphate limitation KW - Phytoplankton Y1 - 2018 U6 - https://doi.org/10.1016/j.scitotenv.2018.01.188 SN - 0048-9697 SN - 1879-1026 VL - 626 SP - 1342 EP - 1349 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Clegg, Mark R. A1 - Wacker, Alexander A1 - Spijkerman, Elly T1 - Phenotypic Diversity and Plasticity of Photoresponse Across an Environmentally Contrasting Family of Phytoflagellates JF - Frontiers in plant science : FPLS N2 - 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. KW - photoresponse KW - behaviour KW - physiology KW - composition KW - photosynthesis KW - acclimation KW - Chlamydomonas KW - ecophysiology Y1 - 2021 U6 - https://doi.org/10.3389/fpls.2021.707541 SN - 1664-462X IS - 12 PB - Frontiers Media CY - Lausanne ER -