Unicellular Alga Research Articles

The response of LncRNAs associated with photosynthesis-and pigment synthesis-related genes to green light in Chlamydomonas reinhardtii.

The quality of light is an important abiotic factor that affects the growth and development of green plants. Ultraviolet, red, blue, and far-red light all have demonstrated roles in regulating green plant growth and development, as well as light morphogenesis. However, the mechanism underlying photosynthetic organism responses to green light throughout the life of them are not clear. In this study, we exposed the unicellular green alga Chlamydomonas reinhardtii to green light and analyzed the dynamics of transcriptome changes. Based on the whole transcriptome data from C. reinhardtii, a total of 9974 differentially expressed genes (DEGs) were identified under green light. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that these DEGs were mainly related to "carboxylic acid metabolic process," "enzyme activity," "carbon metabolism," and "photosynthesis and other processes." At the same time, 253 differentially expressed long non-coding RNAs (DELs) were characterized as green light responsive. We also made a detailed analysis of the responses of photosynthesis- and pigment synthesis-related genes in C. reinhardtii to green light and found that these genes exhibited obvious dynamic expression. Lastly, we constructed a co-expression regulatory network, comprising 49 long non-coding RNAs (lncRNAs) and 20 photosynthesis and pigment related genes, of which 9 mRNAs were also the predicted trans/cis-targets of 8 lncRNAs, these results suggested that lncRNAs may affect the expression of mRNAs related to photosynthesis and pigment synthesis. Our findings give a preliminary explanation of the response mechanism of C. reinhardtii to green light at the transcriptional level.

The first arctic strain of Rhizochromulina: Morphology, ultrastructure, and position in the evolutionary tree of the order Rhizochromulinales (Heterokontophyta, Dictyochophyceae)

Rhizochromulina marina is a unicellular amoeboid alga capable of forming flagellate cells; it is a single validly named species in the genus. Besides, there are numerous environmental sequences and undescribed strains designated as Rhizochromulina sp. or R. marina. The biogeography of the genus is understudied: rhizochromulines from the Indian, Southern, and Arctic Oceans are unknown. Here, we present the description of Rhizochromulina sp. B44, which was for the first time isolated from an arctic habitat. Biofilms of this microalga grow at the bottom of a culture vessel, where neighbouring amoeboid cells form associations through a common network of pseudopodia, i.e. meroplasmodia. Pseudopodia branch, anastomose mainly during meroplasmodia formation, and are supported by microtubules that arise from the perinuclear zone. Actin filaments are localized in the cytoplasm and can be revealed only near the bases of pseudopodia. We succeeded in inducing the transformation of amoeboid cells into flagellates using a prolonged agitation of cultures. Morphological and molecular analyses revealed that the studied strain is most closely related to the type strain of R. marina. At the same time, 18S rDNA sequences of early branching-off rhizochromulinids differ significantly from Rhizochromulina sp. B44, suggesting a high divergence at the genus level.

Exploring the Diversity of Plant-Associated Viruses and Related Viruses in Riverine Freshwater Samples Collected in Berlin, Germany.

Plant-infecting RNA viruses from 30 families and floating genera, as well as a great number of uncultured as yet-unclassified plant-associated viruses have been described. Even so, the plant RNA virosphere is still underexplored. RNA extracted from enriched virus particles of 50 L water samples from the Teltow Canal and the Havel River in Berlin, Germany, was sequenced using Illumina next-generation sequencing. Sequences were searched for plant viruses with BLAST and DIAMOND. Phylogenetic analyses were conducted with IQ-TREE 2. Altogether, 647 virus sequences greater than 1 kb were detected and further analyzed. These data revealed the presence of accepted and novel viruses related to Albetovirus, Alphaflexiviridae, Aspiviridae, Bromoviridae, Endornaviridae, Partitiviridae, Potyviridae, Solemoviridae, Tombusviridae and Virgaviridae. The vast majority of the sequences were novel and could not be taxonomically assigned. Several tombus- and endorna-like viruses make use of alternative translation tables that suggest unicellular green algae, ciliates, or diplomonades as their hosts. The identification of 27 albeto-like satellite viruses increases available sequence data five-fold. Sixteen new poty-like viruses align with other poty-like viruses in a link that combines the Astroviridae and Potyviridae families. Further, the identification of viruses with peptidase A6-like and peptidase A21-like capsid proteins suggests horizontal gene transfer in the evolution of these viruses.

Hidden in plain sight: The invasive macroalga Caulerpa prolifera evades detection by environmental DNA methods

AbstractEnvironmental managers need a rapid and cost‐effective monitoring tool for tracking the spread of invasive species, particularly at the onset of introduction. The macroalgae Caulerpa prolifera is considered an invasive species outside its native range, colonizing large patches of seafloor, reducing native species, and altering ecosystem functioning. Here, we developed a droplet digital PCR assay for detection of C. prolifera from environmental DNA seawater samples using the internal transcribed spacer (ITS) region. While the assay itself was confirmed to be highly efficient, we discovered concentrations of C. prolifera eDNA were present below detectable levels in the water column surrounding an outbreak. To understand why, we conducted tank‐based experiments for two California invasive algae species, Caulerpa prolifera and Sargassum horneri. The steady‐state eDNA concentration (eDNA copies/ gram of biomass detected) of C. prolifera was found to be two orders of magnitude lower than S. horneri. A meta‐analysis of steady‐state concentrations reported in the literature showed a remarkable range from ~104–1011 (copies/g), revealing C. prolifera to have the lowest recorded steady‐state concentrations of eDNA of any known species. We attribute C. prolifera's low steady‐state eDNA concentration to its unique biology as a unicellular macroscopic algae which reduces the possible modes of eDNA release compared to similarly sized multicellular organisms. Critically our results demonstrate the potential limits of eDNA approaches, the influence of shedding rates in the reliability of species detections, and the vital importance of benchmarking and validating eDNA assays in both field and laboratory settings.

Activation of Immune and Antiviral Effects by Euglena Extracts: A Review.

Influenza is an acute respiratory illness caused by influenza virus infection, which is managed using vaccines and antiviral drugs. Recently, the antiviral effects of plants and foods have gained attention. Euglena is a motile unicellular alga and eukaryotic photosynthetic microorganism. It has secondary chloroplasts and is a mixotroph able to feed by photosynthesis or phagocytosis. This review summarizes the influenza treatment effects of Euglena from the perspective of a functional food that is attracting attention. While it has been reported that Euglena contributes to suppressing blood sugar levels and ameliorates symptoms caused by stress by acting on the autonomic nervous system, the immunostimulatory and antiviral activities of Euglena have also been reported. In this review, I focused on the immunostimulation of antiviral activity via the intestinal environment and the suppression of viral replication in infected cells. The functions of specific components of Euglena, which also serves as the source of a wide range of nutrients such as vitamins, minerals, amino acids, unsaturated fatty acids, and β-1,3-glucan (paramylon), are also reviewed. Euglena has animal and plant properties and natural compounds with a wide range of functions, providing crucial information for improved antiviral strategies.

Utilizing high-resolution ribosome profiling for the global investigation of gene expression in Chlamydomonas.

Ribosome profiling (Ribo-seq) is a powerful method for the deep analysis of translation mechanisms and regulatory circuits during gene expression. Extraction and sequencing of ribosome-protected fragments (RPFs) and parallel RNA-seq yields genome-wide insight into translational dynamics and post-transcriptional control of gene expression. Here, we provide details on the Ribo-seq method and the subsequent analysis with the unicellular model alga Chlamydomonas reinhardtii (Chlamydomonas) for generating high-resolution data covering more than 10 000 different transcripts. Detailed analysis of the ribosomal offsets on transcripts uncovers presumable transition states during translocation of elongating ribosomes within the 5' and 3' sections of transcripts and characteristics of eukaryotic translation termination, which are fundamentally distinct for chloroplast translation. In chloroplasts, a heterogeneous RPF size distribution along the coding sequence indicates specific regulatory phases during protein synthesis. For example, local accumulation of small RPFs correlates with local slowdown of psbA translation, possibly uncovering an uncharacterized regulatory step during PsbA/D1 synthesis. Further analyses of RPF distribution along specific cytosolic transcripts revealed characteristic patterns of translation elongation exemplified for the major light-harvesting complex proteins, LHCs. By providing high-quality datasets for all subcellular genomes and attaching our data to the Chlamydomonas reference genome, we aim to make ribosome profiles easily accessible for the broad research community. The data can be browsed without advanced bioinformatic background knowledge for translation output levels of specific genes and their splice variants and for monitoring genome annotation.

Nabumetone and flufenamic acid pose a serious risk to aquatic plants: A study with Chlamydomonas reinhardtii as a model organism

The aquatic environment is constantly under threat due to the release of numerous pollutants. Among them, pharmaceuticals constitute a huge and diverse group. Non-steroidal anti-inflammatory drugs (NSAIDs) are increasingly found in water bodies, but knowledge about their potential toxicity is still low. In particular, there is a lack of information about their influences on aquatic plants and algae. We estimated the susceptibility of the microalgae Chlamydomonas reinhardtii to nabumetone (NBT) and flufenamic acid (FFA), focusing on photosynthesis. Due to the differences in the structures of these compounds, it was assumed that these drugs would have different toxicities to the tested green algae. The hypothesis was confirmed by determining the effective concentration values, the intensity of photosynthesis, the intensity of dark respiration, the contents of photosynthetic pigments, the fluorescence of chlorophyll a in vivo (OJIP test), and cell ultrastructure analysis. Assessment of the toxicity of the NSAIDs was extended by the calculation of an integrated biomarker response index (IBR), which is a valuable tool in ecotoxicological studies. The obtained results indicate an over six times higher toxicity of NBT compared to FFA. After analysis of the chlorophyll a fluorescence in vivo, it was found that NBT inhibited electron transport beyond the PS II. FFA, unlike NBT, lowered the intensity of photosynthesis, probably transforming some reaction centers into “silent centers”, which dissipate energy as heat. The IBR estimated based on photosynthetic parameters suggests that the toxic effect of FFA results mainly from photosynthesis disruption, whereas NBT significantly affects other cellular processes. No significant alteration in the ultrastructure of treated cells could be seen, except for changes in starch grain number and autophagic vacuoles that appeared in FFA-treated cells. To the best of our knowledge, this is the first work reporting the toxic effects of NBT and FFA on unicellular green algae.

Complete mitochondrial genome and phylogenetic analysis of the marine microalga Symbiochlorum hainanensis (Ulvophyceae, Chlorophyta)

Symbiochlorum hainanensis Gong et al. (2018). is a unicellular green alga belonging to Ulvophyceae, Chlorophyta, and considered as an important species in coral-algae symbiont areas. In this study, we revealed firstly the mitochondrial genome sequence of the S. hainanensis. This mitochondrial genome was a circular DNA molecule of 59,508 bp, including 24 transfer RNA genes, 3 ribosomal RNA genes, and 31 protein-coding genes. The GC content of the genome was 35.4%. The phylogenetic tree suggested that S. hainandiae was a sister to the OUU clade within the class Ulvophyceae. The mitochondrial genome structure and gene content of S. hainanensis supported that S. hainanensis was a new unidentified green alga in Ulvophyceae.

The effects of sex on extinction dynamics of Chlamydomonas reinhardtii depend on the rate of environmental change.

The continued existence of sex, despite many the costs it entails, still lacks an adequate explanation, as previous studies demonstrated that the effects of sex are environment-dependent: sex enhances the rate of adaptation in changing environments, but the benefits level off in benign conditions. To the best of our knowledge, the potential impact of different patterns of environmental change on the magnitude of these benefits received less attention in theoretical studies. In this paper, we begin to explore this issue by examining the effect of the rate of environmental deterioration (negatively correlated with population survival rate), on the benefits of sex. To investigate the interplay of sex and the rate of environmental deterioration, we carried out a long-term selection experiment with a unicellular alga (Chlamydomonas reinhardtii), by manipulating mode of reproduction (asexual, facultative or obligate sexual) and the rate of environmental deterioration (an increase of salt concentration). We monitored both the population size and extinction dynamics. The results revealed that the relative advantage of sex increased at the intermediate rate and plateaued at the highest rate of environmental deterioration. Obligate sexual populations had the slowest extinction rate under the intermediate rate of environmental deterioration, while facultative sexuality was favoured under the high rate-treatment. To the best of our knowledge, our study is the first to demonstrate that the interplay of sex and the rate of environmental deterioration affects the probability of survival, which indicates that mode of reproduction may be an important determinant of survival of the anthropogenic-induced environmental change.

Ecological and Geochemical Assessment of Snow Cover in the Area Affected by the Apatite–Nepheline Production of the Kola Peninsula

The chemical composition of the snow cover in the area of industrial development of the apatite–nepheline deposit is analyzed to estimate the ecological and geochemical environmental impact of the mining enterprise. It has been established that the snow of the studied area of the Khibiny is enriched in Cl– and Na+ ions (on average 38 and 41 µeq/L), and relations between basic ions (Cl– and Na+ Ca2+ K+ = Mg2+) and mineralization value (from 1.7 to 6.4 mg/L) are typical for precipitates in the coastal regions of the northern European Russia. The average content of total nitrogen and phosphorus in the snow of the impact zone is 495 and 26 μg/L, respectively, which is 3 and 5 times higher than in the background zone. This is explained by their influx into the atmosphere with dust emissions from the mining enterprise. The content of organic matter (CODMn and TOC 5.5 and 5.8 mg/L) in the snow of the impact zone is about two times higher than in the snow of the background zone and in the water of the Khibiny water bodies. Probably, the elevated content of organic matter in the snow is associated with the supply of organic substances-reagents from the tailing dump, which are used to obtain apatite concentrate, as well as the intensive growth of unicellular green algae Chlamydomonas nivalis (Bauer) Wille under conditions of an increased content of nutrients and long daylight hours. The concentrations of a number of heavy metals (Zn, Mn, Cu, Cr, Pb, Cd) in the snow of the impact zone exceed their contents in the water of water body of the impact zone (13.4, 5.4, 3.8, 0.8, 0.65, 0.035 μg/L, respectively). These metals enter the snow as a part of dust emissions from the mine, and as polluted air masses from the industrial regions of Eurasia.

Efficient assays to quantify the life history traits of algal viruses.

Viruses play a crucial role in microbial ecosystems by liberating nutrients and regulating the growth of their hosts. These effects are governed by viral life history traits, i.e., by the traits determining viral reproduction and survival. Understanding these traits is essential to predicting viral effects, but measuring them is generally labor intensive. In this study, we present efficient methods to quantify the full life cycle of lytic viruses. We developed these methods for viruses infecting unicellular Chlorella algae but expect them to be applicable to other lytic viruses that can be quantified by flow cytometry. By making viral phenotypes accessible, our methods will support research into the diversity and ecological effects of microbial viruses.

Nanophytosome formulation of β-1,3-glucan and Euglena gracilis extract for drug delivery applications

Euglena gracilis (EG) is a unicellular freshwater alga known for its high β-1,3-glucan (BG) content with well-known biological properties and immune response. The high molecular weight structure of BG traditionally poses a challenge in terms of its size and absorption. Therefore, the aim of this study was to develop a novel drug delivery mechanism of BG and EG to nanophytosomes (NPs) by converting the heavy molecular weight of BG and EG into lipid phosphatidylcholine (PC), which plays an important role in improving their bioavailability and entrapment in captivity. The BG and EG NPs were developed by the solvent evaporation method while varying time and temperature to optimize their drug delivery ability. The size of BG-PC and EG-PC obtained by the Dynamic Light Scattering (DLS) method was 134.62 and 158.38 nm, respectively. Chemical (Fourier Transform Infra-Red) and structural (X-Ray Diffraction) characterization of NPs improved the binding capacity and the amorphous nature of both NPs. The shape of the NPs by Scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) revealed their spherical, vesicular nature. The encapsulation efficiency of BG-PC and EG-PC was 82 ± 1.62 % and 87 ± 3.22 %, respectively, which improves the bioavailability. The developed methodology has thus proven effective in synthesizing BG-PC and EG-PC, which may be useful as NP drug delivery carriers. Future research could demonstrate the safety and effectiveness of long-term storage conditions for medical and pharmaceutical applications.•Nanophytosomes are tailored in size, shape and composition to optimize the delivery of phytochemicals/phytocompounds through nanoscale size and surface modification for better physiological absorption.•Nanophytosomes increase the stability of phytochemicals/phytocompounds and protect them from degradation due to heat or chemical reactions, leading to longer shelf life and improved therapeutic efficacy.•In this method, optimal conditions were created for the formation of β-1,3-glucan and Euglena gracilis extract nanophytosomes for successful development of drug delivery system that can effectively deliver bioactive compounds.

A facile Agrobacterium-mediated transformation method for the model unicellular green algae Chlamydomonas reinhardtii

A reliable and simple Agrobacterium-mediated transformation system for the unicellular green algae model organism Chlamydomonas reinhardtii has been developed. The protocol has been successfully employed with both neomycin phosphotransferase II (nptII) and the phleomycin resistance (bleI) genes coupled with the selective agents paromomycin and zeocin, respectively. A set of binary vectors were assembled that carry the selectable marker cassettes under control either of the Rbcs2 alone or fused to the HSP270A leader sequence, PsaD, or ß-tubulin2 promoters. The corresponding T-DNA elements also harbored a cassette with a codon-optimized version of yellow fluorescence protein (YFP) under control of the Rbcs2 promoter in which the YFP open reading frame was interrupted with the first intron of Rbcs2 to prevent expression in Agrobacterium tumefaciens. The resultant binary vectors were introduced into A. tumefaciens strain C58C1/pMP90, and the derived transconjugants were used for transformation studies with the walled C. reinhardtii strain CC124. Estimated transformation frequencies ranged from 0.09 to 2.86 colonies per 106 cells inoculated. Molecular characterizations on a subset of the transgenic lineages revealed that most of the transgenic events harbored single locus insertions. Moreover, sequencing of captured junction fragments about the T-DNA insertion site showed that minimal disruption of the C. reinhardtii genome occurred. However, the transgenic lineages often harbored truncated T-DNA regions within the non-selectable marker gene cassettes.

Structures, characteristics and functions of fructose-1,6-bisphosphate aldolase in various tissues

Aldolase exhibits multiple functions in a variety of organisms, including fungi, unicellular algae and plants, and so on. Furthermore, different isoforms of fructose 1,6-bisphosphate aldolase (FBA) exhibit significantly different characteristics and functions. This review describes the structures, characteristics, and functions of FBAs derived from diverse organisms, including FBA II in pathogen microorganisms, as well as FBA I mainly in unicellular algae and plants. Differences between FBA and FBP aldolase-phosphatase bifunctional enzyme (FBA/P) are also discussed. Finally, we suggest several potential research questions regarding the functions of FBA in higher plants.

A cell-based model for size control in the multiple fission alga Chlamydomonas reinhardtii

Understanding how population-size homeostasis emerges from stochastic individual cell behaviors remains a challenge in biology.1,2,3,4,5,6,7 The unicellular green alga Chlamydomonas reinhardtii (Chlamydomonas) proliferates using a multiple fission cell cycle, where a prolonged G1 phase is followed by n rounds of alternating division cycles (S/M) to produce 2n daughters. A "Commitment" sizer in mid-G1 phase ensures sufficient cell growth before completing the cell cycle. A mitotic sizer couples mother-cell size to division number (n) such that daughter size distributions are uniform regardless of mother size distributions. Although daughter size distributions were highly robust to altered growth conditions, ∼40% of daughter cells fell outside of the 2-fold range expected from a "perfect" multiple fission sizer.7,8 A simple intuitive power law model with stochastic noise failed to reproduce individual division behaviors of tracked single cells. Through additional iterative modeling, we identified an alternative modified threshold (MT) model, where cells need to cross a threshold greater than 2-fold their median starting size to become division-competent (i.e., Committed), after which their behaviors followed a power law model. The Commitment versus mitotic size threshold uncoupling in the MT model was likely a key pre-adaptation in the evolution of volvocine algal multicellularity. A similar experimental approach was used in size mutants mat3/rbr and dp1 that are, respectively, missing repressor or activator subunits of the retinoblastoma tumor suppressor complex (RBC). Both mutants showed altered relationships between Commitment and mitotic sizer, suggesting that RBC functions to decouple the two sizers.

Dynamics of Phytoplankton Species Structure in the Kurile Lake (Kamchatka Peninsula)

The species structure of the dominant community of unicellular algae inhabiting the spawning and feeding salmon lake was studied in the Kurile Lake (southern Kamchatka peninsula). It has been shown that this structure is well described by the Motomura model. The geometric progression coefficient does not differ significantly from the value prescribed by the model, which is based on the stochastic remainder hypothesis. Several integral indicators characterizing the species structure have been calculated. The study has revealed some typical and unusual correlations between these integral indices. Inter-annual and intra-annual cyclical fluctuations in the value of indicators have been found.

Evaluation the toxic effect of copper ions on the condition indices of benthic diatom Actinocyclus subtilis (W.Gregory) Ralfs 1861 in the experiment

Introduction. Pollution of marine coastal areas lead to the relevance of environmental monitoring including application of biotesting methods based on- the cultures of unicellular algae. Microalgae have different species-specific resistance to pollutants that expands application of different species as bioindicators of marine pollution.
 The aim of the study was to determine the threshold concentration of copper ions (Cu2+) for the survival and increase in the cells number of benthic diatom Actinocyclus subtilis (W.Gregory) Ralfs 1861 (Bacillariophyta) under the wide range of toxicant concentrations during 10-day toxicological experiments.
 Material and methods. The response of strain culture of the benthic diatom A. subtilis to various concentrations of copper sulfate (ranged from 16 to 1024 μg/l in terms of Cu2+ ions) was studied. In accordance with the previously developed protocol, the following indices were evaluated: alterations in the absolute number and proportion (%) of alive cells in the test-culture, as well as the specific growth rate in the number of A. subtilis cells at different concentrations of toxicant. Counting of alive and dead cells was carried out by micrographs taken for 12–15 random viewing fields under Nikon Eclipse inverted light microscope.
 Results. It was found that in the control and at concentration of copper ions 16 μg/l, the increase in the absolute number of cells in culture is described by sigmoid response curve. At the control еhe exponential growth phase occurs on days 5–7 and at concentration of 16 µg/l on days 3–5 of the experiment. The threshold concentration of copper ions (32 μg/l) which is critical for the survival of A. subtilis was determined, which is 3–7 times lower than threshold level for other benthic diatom species. At concentration of 32 µg/l, the phases of acceleration and exponential growth on the abundance curve are absent. The proportion of living cells in the culture decreases to 80% of the control level on day 3 and to 39% by day 10. At Cu2+ concentrations of 64 µg/l and above, sharp inhibition and death of culture is observed as early as 1–3 days. A positive specific growth rate of A. subtilis culture was revealed in the period of 1–5 days at copper concentration of 16 and 
 32 µg/l, and at concentration of 64 µg/l and higher the culture dies off. Negative values of the specific growth rate for all concentrations of the toxicant within the period of 5–10 days were obtained.
 Limitations. By the results of 10-day experiments the effect of 8 concentrations of copper sulfate on the culture of marine benthic diatom A. subtilis was studied. Three replicates in each concentration and exposure time were measured (1350 measurements in total), which is sufficient sampling for statistically reliable determination of the threshold values of copper ion toxicity for given test object.
 Conclusion. Considering the results obtained, the benthic diatom A. subtilis is highly sensitive to copper ions impact and can be recommended as new test-object for toxicology, as well as for application in monitoring of marine water areas subject to technogenic pollution.

Moderate high temperature is beneficial or detrimental depending on carbon availability in the green alga Chlamydomonas reinhardtii.

High temperatures impair plant growth and reduce agricultural yields, but the underlying mechanisms remain elusive. The unicellular green alga Chlamydomonas reinhardtii is an excellent model to study heat responses in photosynthetic cells due to its fast growth rate, many similarities in cellular processes to land plants, simple and sequenced genome, and ample genetic and genomics resources. Chlamydomonas grows in light by photosynthesis and with externally supplied acetate as an organic carbon source. Understanding how organic carbon sources affect heat responses is important for the algal industry but remains understudied. We cultivated wild-type Chlamydomonas under highly controlled conditions in photobioreactors at 25 °C (control), 35 °C (moderate high temperature), or 40 °C (acute high temperature) with or without constant acetate supply for 1 or 4 day. Treatment at 35 °C increased algal growth with constant acetate supply but reduced algal growth without sufficient acetate. The overlooked and dynamic effects of 35 °C could be explained by induced acetate uptake and metabolism. Heat treatment at 40 °C for more than 2 day was lethal to algal cultures with or without constant acetate supply. Our findings provide insights to understand algal heat responses and help improve thermotolerance in photosynthetic cells.

An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression.

Riboregulators such as riboswitches and RNA thermometers provide simple, protein-independent tools to control gene expression at the post-transcriptional level. In bacteria, RNA thermometers regulate protein synthesis in response to temperature shifts. Thermometers outside of the bacterial world are rare, and in organellar genomes, no RNA thermometers have been identified to date. Here we report the discovery of an RNA thermometer in a chloroplast gene of the unicellular green alga Chlamydomonas reinhardtii. The thermometer, residing in the 5' untranslated region of the psaA messenger RNA forms a hairpin-type secondary structure that masks the Shine-Dalgarno sequence at 25°C. At 40°C, melting of the secondary structure increases accessibility of the Shine-Dalgarno sequence to initiating ribosomes, thus enhancing protein synthesis. By targeted nucleotide substitutions and transfer of the thermometer into Escherichia coli, we show that the secondary structure is necessary and sufficient to confer the thermometer properties. We also demonstrate that the thermometer provides a valuable tool for inducible transgene expression from the Chlamydomonas plastid genome, in that a simple temperature shift of the algal culture can greatly increase recombinant protein yields.

SNG100, a novel topical treatment for moderate atopic dermatitis, in patients aged 6years or older: A randomised, double-blind, active-controlled trial.

Atopic dermatitis (AD) is one of the most common inflammatory skin diseases. It is associated with significant itch and impaired quality of life. Systemic treatments are efficient but associated with side effects. Novel topical treatments with a favourable safety profile are needed. SNG100 is a novel composition of hydrocortisone 1% in a cream base comprising sulphated polysaccharide (SPS; extracted from in-house cultivated Porphyridium Cruentum unicellular algae), a well-known hydrating, moisturising and a skin barrier repairing agent. To assess the safety, usability and efficacy of SNG100 cream in patients aged ≥6years with moderate AD. In this proof of concept phase I, double-blind, randomised trial, participants received one of three treatments for 14days: SNG100 twice daily (BID), hydrocortisone 1% BID or mometasone furoate once daily (QD). The primary endpoint was the safety and tolerability of SNG100 cream compared to hydrocortisone 1% and mometasone furoate. The secondary endpoint was the subject's usability of SNG100. Exploratory efficacy endpoints included percent change from baseline in SCOring AD (SCORAD), Eczema Area and Severity Index, Patient-Oriented Eczema Measure, Dermatology Life Quality Index, pruritus Numerical Rating Score (NRS), peak pruritus-NRS and Investigator's Global Assessment. Subjects were also followed up without any treatment for additional 14days. Overall, 66 participants were screened, and 60 patients were randomised. SNG100 demonstrated a high safety profile, similar to marketed products hydrocortisone 1% and mometasone furoate 0.1%, with no unanticipated drug safety related events. SNG100 and mometasone furoate 0.1% cream achieved almost similar and statistically significant greater percentage reductions from baseline in SCORAD as compared to hydrocortisone 1% cream. SNG100 demonstrated significant improvement in NRS as compared to hydrocortisone 1% cream. Remarkably, SNG100 led to a lasting effect with only 29.4% of subjects returning to IGA3 during the follow-up period compared to 50% and 38.9% in the hydrocortisone 1% and in mometasone furoate treatment arms, respectively. Topical SNG100 is an effective, safe, and well-tolerated innovative treatment for moderate AD. Trial registration number: NCT04615962 (Topical Cream SNG100 for Treatment in Moderate AD Subjects).