Nitrogen is one of the most important nutrient sources for the growth of microalgae. We studied the effects of nitrogen starvation on the growth responses, biochemical composition, and fatty acid profile of Dunaliella tertiolecta, Phaeodactylum tricornutum, and Nannochloropsis oculata. The lack of nitrogen caused changes in carbohydrate, protein, lipid, and fatty acid composition in all examined microalgae. The carbohydrate content increased 59% in D. tertiolecta, while the lipid level increased 139% in P. tricornutum under nitrogen stress conditions compared to the control groups. Nitrogen starvation increased the oligosaccharide and polysaccharide contents of D. tertiolecta 4.1-fold and 3.6-fold, respectively. Furthermore, triacylglycerol (TAG) levels in N. oculata and P. tricornutum increased 2.3-fold and 7.4-fold, respectively. The dramatic increase in the amount of TAG is important for the use of these microalgae as raw materials in biodiesel. Nitrogen starvation increased the amounts of oligosaccharides and polysaccharides of D. tertiolecta, while increased eicosapentaenoic acid (EPA) in N. oculata and docosahexaenoic acid (DHA) content in P. tricornutum. The amount of polyunsaturated fatty acids (PUFAs), EPA, DHA, oligosaccharides, and polysaccharides in microalgal species can be increased without using the too costly nitrogen source in the culture conditions, which can reduce the most costly of living feeding.

• Klíčová slova
• Biochemical composition, Fatty acid profile, Microalgae, Nitrogen starvation,
• MeSH
• dusík * metabolismus   MeSH
• mastné kyseliny * metabolismus   analýza   MeSH
• mikrořasy * metabolismus   růst a vývoj   MeSH
• triglyceridy metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dusík * MeSH
• mastné kyseliny * MeSH
• triglyceridy MeSH

The globally distributed marine alga Emiliania huxleyi has cooling effect on the Earth's climate. The population density of E. huxleyi is restricted by Nucleocytoviricota viruses, including E. huxleyi virus 201 (EhV-201). Despite the impact of E. huxleyi viruses on the climate, there is limited information about their structure and replication. Here, we show that the dsDNA genome inside the EhV-201 virion is protected by an inner membrane, capsid, and outer membrane. EhV-201 virions infect E. huxleyi by using fivefold vertices to bind to and fuse the virus' inner membrane with the cell plasma membrane. Progeny virions assemble in the cytoplasm at the surface of endoplasmic reticulum-derived membrane segments. Genome packaging initiates synchronously with the capsid assembly and completes through an aperture in the forming capsid. The genome-filled capsids acquire an outer membrane by budding into intracellular vesicles. EhV-201 infection induces a loss of surface protective layers from E. huxleyi cells, which enables the continuous release of virions by exocytosis.

• MeSH
• Haptophyta * metabolismus   MeSH
• Phycodnaviridae * genetika   MeSH
• podnebí MeSH
• virion MeSH
• viry * MeSH
• Publikační typ
• časopisecké články MeSH

The main entrance point of highly toxic organic Hg forms, including methylmercury (MeHg), into the aquatic food web is phytoplankton, which is greatly represented by various natural microalgal species. Processes associated with MeHg fate in microalgae cells such as uptake, effects on cells and toxicity, Hg biotransformation, and intracellular stability are detrimental to the process of further biomagnification and, as a consequence, have great importance for human health. The study of MeHg uptake and distribution in cultures of marine halophile Dunaliella salina and freshwater acidophilic alga Coccomyxa onubensis demonstrated that most of the MeHg is imported inside the cell, while cell surface adhesion is insignificant. Almost all MeHg is removed from the culture medium after 72 h. Significant processes in rapid MeHg removal from liquid medium are its abiotic photodegradation and volatilization associated with algal enzymatic activity. The maximum intracellular accumulation for both species was in 80 nM MeHg-exposed cultures after 24 h of exposure for D. salina (from 27 to 34 µg/gDW) and at 48 h for C. onubensis (up to 138 µg/gDW). The different Hg intakes in these two strains could be explained by the lack of a rigid cell wall in D. salina and the higher chemical ability of MeHg to pass through complex cell wall structures in C. onubensis. Electron microscopy studies on the ultrastructure of both strains demonstrated obvious microvacuolization in the form of many very small vacuoles and partial cell membrane disruption in 80 nM MeHg-exposed cultures. Results further showed that Coccomyxa onubensis is a good candidate for MeHg-contaminated water reclamation due to its great robustness at nanomolar concentrations of MeHg coupled with its very high intake and almost complete Hg removal from liquid medium at the MeHg levels tested.

• Klíčová slova
• mercury contamination, mercury toxicity, methylmercury, microalgae,
• Publikační typ
• časopisecké články MeSH

Most organisms on Earth are affected by periodic changes in their environment. The circadian clock is an endogenous device that synchronizes behavior, physiology, or biochemical processes to an approximately 24-hour cycle, allowing organisms to anticipate the periodic changes of day and night. Although circadian clocks are widespread in organisms, the actual molecular components differ remarkably among the clocks of plants, animals, fungi, and prokaryotes. Chromera velia is the closest known photosynthetic relative of apicomplexan parasites. Formation of its motile stage, zoospores, has been described as associated with the light part of the day. We examined the effects on the periodic release of the zoospores under different light conditions and investigated the influence of the spectral composition on zoosporogenesis. We performed a genomic search for homologs of known circadian clock genes. Our results demonstrate the presence of an almost 24-hour free-running cycle of zoosporogenesis. We also identified the blue light spectra as the essential compound for zoosporogenesis. Further, we developed a new and effective method for zoospore separation from the culture and estimated the average motility speed and lifespan of the C. velia zoospores. Our genomic search identified six cryptochrome-like genes, two genes possibly related to Arabidopsis thaliana CCA/LHY, whereas no homolog of an animal, cyanobacterial, or fungal circadian clock gene was found. Our results suggest that C. velia has a functional circadian clock, probably based mainly on a yet undefined mechanism.

• Klíčová slova
• Chromera velia, apicomplexa, circadian clock, cryptochrome, zoospore formation,
• Publikační typ
• časopisecké články MeSH

Nitrogen fixing plankton provide nitrogen to fuel marine ecosystems and biogeochemical cycles but the factors that constrain their growth and habitat remain poorly understood. Here we investigate the importance of metabolic specialization in unicellular diazotroph populations, using laboratory experiments and model simulations. In clonal cultures of Crocosphaera watsonii and Cyanothece sp. spiked with 15N2, cellular 15N enrichment developed a bimodal distribution within colonies, indicating that N2 fixation was confined to a subpopulation. In a model of population metabolism, heterogeneous nitrogen (N2) fixation rates substantially reduce the respiration rate required to protect nitrogenase from O2. The energy savings from metabolic specialization is highest at slow growth rates, allowing populations to survive in deeper waters where light is low but nutrients are high. Our results suggest that heterogeneous N2 fixation in colonies of unicellular diazotrophs confers an energetic advantage that expands the ecological niche and may have facilitated the evolution of multicellular diazotrophs.

• MeSH
• biologická evoluce * MeSH
• biologické modely MeSH
• Cyanothece růst a vývoj   metabolismus   MeSH
• dusík metabolismus   MeSH
• ekosystém MeSH
• energetický metabolismus * MeSH
• fixace dusíku * MeSH
• fyziologická adaptace MeSH
• počítačová simulace MeSH
• sinice růst a vývoj   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dusík MeSH

In this paper, we describe a novel bacteriophagous biflagellate, Cafileria marina with two smooth flagellae, isolated from material collected from a rock surface in the Kvernesfjorden (Norway). This flagellate was characterized by scanning and transmission electron microscopy, fluorescence, and light microscopy. The sequence of the small subunit ribosomal RNA gene (18S) was used as a molecular marker for determining the phylogenetic position of this organism. Apart from the nuclear ribosomal gene, the whole mitochondrial genome was sequenced, assembled, and annotated. Morphological observations show that the newly described flagellate shares key ultrastructural characters with representatives of the family Bicosoecida (Heterokonta). Intriguingly, mitochondria of C. marina frequently associate with its nucleus through an electron-dense disc at the boundary of the two compartments. The function of this association remains unclear. Phylogenetic analyses corroborate the morphological data and place C. marina with other sequence data of representatives from the family Bicosoecida. We describe C. marina as a new species from a new genus in this family.

• Klíčová slova
• bicosoecida, flagellar apparatus, heterokonta, heterotrophic nano-flagellate, new genus, phylogeny,
• Publikační typ
• časopisecké články MeSH

Photoprotective non-photochemical quenching (NPQ) represents an effective way to dissipate the light energy absorbed in excess by most phototrophs. It is often claimed that NPQ formation/relaxation kinetics are determined by xanthophyll composition. We, however, found that, for the alveolate alga Chromera velia, this is not the case. In the present paper, we investigated the reasons for the constitutive high rate of quenching displayed by the alga by comparing its light harvesting strategies with those of a model phototroph, the land plant Spinacia oleracea. Experimental results and in silico studies support the idea that fast quenching is due not to xanthophylls, but to intrinsic properties of the Chromera light harvesting complex (CLH) protein, related to amino acid composition and protein folding. The pKa for CLH quenching was shifted by 0.5 units to a higher pH compared with higher plant antennas (light harvesting complex II; LHCII). We conclude that, whilst higher plant LHCIIs are better suited for light harvesting, CLHs are 'natural quenchers' ready to switch into a dissipative state. We propose that organisms with antenna proteins intrinsically more sensitive to protons, such as C. velia, carry a relatively high concentration of violaxanthin to improve their light harvesting. In contrast, higher plants need less violaxanthin per chlorophyll because LHCII proteins are more efficient light harvesters and instead require co-factors such as zeaxanthin and PsbS to accelerate and enhance quenching.

• MeSH
• Alveolata fyziologie   MeSH
• bílkoviny řas metabolismus   MeSH
• fotosyntéza * MeSH
• protony * MeSH
• protozoální proteiny metabolismus   MeSH
• rostlinné proteiny metabolismus   MeSH
• Spinacia oleracea fyziologie   MeSH
• světlosběrné proteinové komplexy metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• bílkoviny řas MeSH
• protony * MeSH
• protozoální proteiny MeSH
• rostlinné proteiny MeSH
• světlosběrné proteinové komplexy MeSH

Diatoms are unicellular algae and important primary producers. The process of carbon fixation in diatoms is very efficient even though the availability of dissolved CO2 in sea water is very low. The operation of a carbon concentrating mechanism (CCM) also makes the more abundant bicarbonate accessible for photosynthetic carbon fixation. Diatoms possess carbonic anhydrases as well as metabolic enzymes potentially involved in C4 pathways; however, the question as to whether a C4 pathway plays a general role in diatoms is not yet solved. While genome analyses indicate that the diatom Phaeodactylum tricornutum possesses all the enzymes required to operate a C4 pathway, silencing of the pyruvate orthophosphate dikinase (PPDK) in a genetically transformed cell line does not lead to reduced photosynthetic carbon fixation. In this study, we have determined the intracellular location of all enzymes potentially involved in C4-like carbon fixing pathways in P. tricornutum by expression of the respective proteins fused to green fluorescent protein (GFP), followed by fluorescence microscopy. Furthermore, we compared the results to known pathways and locations of enzymes in higher plants performing C3 or C4 photosynthesis. This approach revealed that the intracellular distribution of the investigated enzymes is quite different from the one observed in higher plants. In particular, the apparent lack of a plastidic decarboxylase in P. tricornutum indicates that this diatom does not perform a C4-like CCM.

• Klíčová slova
• C4 photosynthesis, Carboxylation, Chloroplast, Decarboxylation, Green fluorescent protein (GFP),
• MeSH
• Arabidopsis fyziologie   MeSH
• fosfoenolpyruvátkarboxylasa klasifikace   metabolismus   MeSH
• fotosyntéza fyziologie   MeSH
• koloběh uhlíku MeSH
• kukuřice setá fyziologie   MeSH
• mitochondrie enzymologie   MeSH
• pyruvátkarboxylasa genetika   metabolismus   MeSH
• regulace genové exprese enzymů fyziologie   MeSH
• regulace genové exprese u rostlin fyziologie   MeSH
• rozsivky enzymologie   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfoenolpyruvátkarboxylasa MeSH
• pyruvátkarboxylasa MeSH

Photosystem I (PSI) is a multi-subunit integral pigment-protein complex that performs light-driven electron transfer from plastocyanin to ferredoxin in the thylakoid membrane of oxygenic photoautotrophs. In order to achieve the optimal photosynthetic performance under ambient irradiance, the absorption cross section of PSI is extended by means of peripheral antenna complexes. In eukaryotes, this role is played mostly by the pigment-protein complexes of the LHC family. The structure of the PSI-antenna supercomplexes has been relatively well understood in organisms harboring the primary plastid: red algae, green algae and plants. The secondary endosymbiotic algae, despite their major ecological importance, have so far received less attention. Here we report a detailed structural analysis of the antenna-PSI association in the stramenopile alga Nannochloropsis oceanica (Eustigmatophyceae). Several types of PSI-antenna assemblies are identified allowing for identification of antenna docking sites on the PSI core. Instances of departure of the stramenopile system from the red algal model of PSI-Lhcr structure are recorded, and evolutionary implications of these observations are discussed.

• Klíčová slova
• Electron microscopy, Light-harvesting complex, Nannochloropsis, Photosystem I, Stramenopila,
• MeSH
• fotosystém I (proteinový komplex) metabolismus   MeSH
• plastidy metabolismus   MeSH
• Rhodophyta metabolismus   MeSH
• spektrofotometrie ultrafialová MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fotosystém I (proteinový komplex) MeSH

Eustigmatophyte algae represent an interesting model system for the study of the regulation of the excitation energy flow due to their use of violaxanthin both as a major light-harvesting pigment and as the basis of xanthophyll cycle. Fluorescence induction kinetics was studied in an oleaginous marine alga Nannochloropsis oceanica. Nonphotochemical fluorescence quenching was analyzed in detail with respect to the state of the cellular xanthophyll pool. Two components of nonphotochemical fluorescence quenching (NPQ), both dependent on the presence of zeaxanthin, were clearly resolved, denoted as slow and fast NPQ based on kinetics of their formation. The slow component was shown to be in direct proportion to the amount of zeaxanthin, while the fast NPQ component was transiently induced in the presence of membrane potential on subsecond timescales. The applicability of these observations to other eustigmatophyte species is demonstrated by measurements of other representatives of this algal group, both marine and freshwater.

• Klíčová slova
• Chl a fluorescence, Eustigmatophyceae, Nannochloropsis, Nonphotochemical quenching, Xanthophyll cycle,
• MeSH
• fluorescence MeSH
• fotosyntéza MeSH
• mořské řasy chemie   MeSH
• Publikační typ
• časopisecké články MeSH