ABSTRACT
Energy status and nutrients regulate photosynthetic protein expression. The unicellular green alga Chromochloris zofingiensis switches off photosynthesis in the presence of exogenous glucose (+Glc) in a process that depends on hexokinase (HXK1). Here, we show that this response requires that cells lack sufficient iron (-Fe). Cells grown in -Fe+Glc accumulate triacylglycerol (TAG) while losing photosynthesis and thylakoid membranes. However, cells with an iron supplement (+Fe+Glc) maintain photosynthesis and thylakoids while still accumulating TAG. Proteomic analysis shows that known photosynthetic proteins are most depleted in heterotrophy, alongside hundreds of uncharacterized, conserved proteins. Photosynthesis repression is associated with enzyme and transporter regulation that redirects iron resources to (a) respiratory instead of photosynthetic complexes and (b) a ferredoxin-dependent desaturase pathway supporting TAG accumulation rather than thylakoid lipid synthesis. Combining insights from diverse organisms from green algae to vascular plants, we show how iron and trophic constraints on metabolism aid gene discovery for photosynthesis and biofuel production.
Subject(s)
Chlorophyta , Glucose , Iron , Lipid Metabolism , Photosynthesis , Triglycerides , Iron/metabolism , Glucose/metabolism , Triglycerides/metabolism , Chlorophyta/metabolism , Chlorophyta/genetics , Thylakoids/metabolism , Proteomics , Hexokinase/metabolism , Hexokinase/genetics , Chlorophyceae/metabolism , Chlorophyceae/geneticsABSTRACT
The colonial green microalga Botryococcus braunii is well known for producing liquid hydrocarbons that can be utilized as biofuel feedstocks. B. braunii is taxonomically classified as a single species made up of three chemical races, A, B, and L, that are mainly distinguished by the hydrocarbons produced. We previously reported a B race draft nuclear genome, and here we report the draft nuclear genomes for the A and L races. A comparative genomic study of the three B. braunii races and 14 other algal species within Chlorophyta revealed significant differences in the genomes of each race of B. braunii. Phylogenomically, there was a clear divergence of the three races with the A race diverging earlier than both the B and L races, and the B and L races diverging from a later common ancestor not shared by the A race. DNA repeat content analysis suggested the B race had more repeat content than the A or L races. Orthogroup analysis revealed the B. braunii races displayed more gene orthogroup diversity than three closely related Chlamydomonas species, with nearly 24-36% of all genes in each B. braunii race being specific to each race. This analysis suggests the three races are distinct species based on sufficient differences in their respective genomes. We propose reclassification of the three chemical races to the following species names: Botryococcus alkenealis (A race), Botryococcus braunii (B race), and Botryococcus lycopadienor (L race).
Subject(s)
Chlorophyta , Genomics , Phylogeny , Genomics/methods , Chlorophyta/genetics , Chlorophyta/classification , Genome, Plant , Hydrocarbons/metabolismABSTRACT
The green algae of the genus Ancylonema, which belong to the zygnematophytes, are prevalent colonizers of glaciers worldwide. They display a striking reddish-brown pigmentation in their natural environment, due to vacuolar compounds related to gallic acid. This pigmentation causes glacier darkening when these algae bloom, leading to increased melting rates. The Ancylonema species known so far are true psychrophiles, which hinders experimental work and limits our understanding of these algae. For instance, the biosynthesis, triggering factors, and biological function of Ancylonema's secondary pigments remain unknown. In this study, we introduce a mesophilic Ancylonema species, A. palustre sp. nov., from temperate moorlands. This species forms the sister lineage to all known psychrophilic strains. Despite its morphological similarity to the latter, it exhibits unique autecological and photophysiological characteristics. It allows us to describe vegetative and sexual cellular processes in great detail. We also conducted experimental tests for abiotic factors that induce the secondary pigments of zygnematophytes. We found that low nutrient conditions combined with ultraviolet B radiation result in vacuolar pigmentation, suggesting a sunscreen function. Our thriving, bacteria-free cultures of Ancylonema palustre will enable comparative genomic studies of mesophilic and extremophilic zygnematophytes. These studies may provide insights into how Ancylonema species colonized the world's glaciers.
Subject(s)
Phylogeny , Pigments, Biological , Vacuoles , Pigments, Biological/metabolism , Vacuoles/metabolism , Chlorophyta/metabolism , Chlorophyta/genetics , Pigmentation , Chlorophyceae/metabolism , Chlorophyceae/geneticsABSTRACT
Marine microbes are important in biogeochemical cycling, but the nature and magnitude of their contributions are influenced by their associated viruses. In the presence of a lytic virus, cells that have evolved resistance to infection have an obvious fitness advantage over relatives that remain susceptible. However, susceptible cells remain extant in the wild, implying that the evolution of a fitness advantage in one dimension (virus resistance) must be accompanied by a fitness cost in another dimension. Identifying costs of resistance is challenging because fitness is context-dependent. We examined the context dependence of fitness costs in isolates of the picophytoplankton genus Micromonas and their co-occurring dsDNA viruses using experimental evolution. After generating 88 resistant lineages from two ancestral Micromonas strains, each challenged with one of four distinct viral strains, we found resistance led to a 46% decrease in mean growth rate under high irradiance and a 19% decrease under low. After a year in culture, the experimentally selected lines remained resistant, but fitness costs had attenuated. Our results suggest that the cost of resistance in Micromonas is dependent on environmental conditions and the duration of population adaptation, illustrating the dynamic nature of fitness costs of viral resistance among marine protists.
Subject(s)
Genetic Fitness , Microalgae , Microalgae/virology , Microalgae/genetics , DNA Viruses/genetics , Chlorophyta/virology , Chlorophyta/geneticsABSTRACT
Chromochloris zofingiensis, a unicellular green alga, is a potential source of natural carotenoids. In this study, the mutant LUT-4 was acquired from the chemical mutagenesis pool of C. zofingiensis strain. The biomass yield and lutein content of LUT-4 reached 9.23 g·L-1, and 0.209% of dry weight (DW) on Day 3, which was 49.4%, and 33% higher than that of wild-type (WT), respectively. The biomass yields of LUT-4 under 100, 300, and 500 µmol/m2/s reached 8.4 g·L-1, 7.75 g·L-1, and 6.6 g·L-1, which was 10.4%, 21%, and 29.6% lower compared with the control, respectively. Under mixotrophic conditions, the lutein yields were significantly higher than that obtained in the control. The light intensity of 300 µmol/m2/s was optimal for lutein biosynthesis and the content of lutein reached 0.294% of DW on Day 3, which was 40.7% more than that of the control. When LUT-4 was grown under 300 µmol/m2/s, a significant increase in expression of genes implicated in lutein biosynthesis, including phytoene synthase (PSY), phytoene desaturase (PDS), and lycopene epsilon cyclase (LCYe) was observed. The changes in biochemical composition, Ace-CoA, pyruvate, isopentenyl pyrophosphate (IPP), and geranylgeranyl diphosphate (GGPP) contents during lutein biosynthesis were caused by utilization of organic carbon. It was thereby concluded that 300 µmol/m2/s was the optimal culture light intensity for the mutant LUT-4 to synthesize lutein. The results would be helpful for the large-scale production of lutein.
Subject(s)
Light , Lutein , Lutein/biosynthesis , Chlorophyta/genetics , Chlorophyta/metabolism , Mutation , Biomass , Carotenoids/metabolismABSTRACT
The globin superfamily of proteins is ancient and diverse. Regular assessments based on the increasing number of available genome sequences have elaborated on a complex evolutionary history. In this review, we present a summary of a decade of advances in characterising the globins of cyanobacteria and green algae. The focus is on haem-containing globins with an emphasis on recent experimental developments, which reinforce links to nitrogen metabolism and nitrosative stress response in addition to dioxygen management. Mention is made of globins that do not bind haem to provide an encompassing view of the superfamily and perspective on the field. It is reiterated that an effort toward phenotypical and in-vivo characterisation is needed to elucidate the many roles that these versatile proteins fulfil in oxygenic photosynthetic microbes. It is also proposed that globins from oxygenic organisms are promising proteins for applications in the biotechnology arena.
Subject(s)
Chlorophyta , Cyanobacteria , Globins , Cyanobacteria/metabolism , Cyanobacteria/genetics , Chlorophyta/metabolism , Chlorophyta/genetics , Globins/genetics , Globins/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Heme/metabolism , Nitrogen/metabolism , PhotosynthesisABSTRACT
In flowering plants, euchromatic transposons are transcriptionally silenced by RNA-directed DNA Methylation, a small RNA-guided de novo methylation pathway. RNA-directed DNA Methylation requires the activity of the RNA Polymerases IV and V, which produce small RNA precursors and noncoding targets of small RNAs, respectively. These polymerases are distinguished from Polymerase II by multiple plant-specific paralogous subunits. Most RNA-directed DNA Methylation components are present in all land plants, and some have been found in the charophytic green algae, a paraphyletic group that is sister to land plants. However, the evolutionary origin of key RNA-directed DNA Methylation components, including the two largest subunits of Polymerase IV and Polymerase V, remains unclear. Here, we show that multiple lineages of charophytic green algae encode a single-copy precursor of the largest subunits of Polymerase IV and Polymerase V, resolving the two presumed duplications in this gene family. We further demonstrate the presence of a Polymerase V-like C-terminal domain, suggesting that the earliest form of RNA-directed DNA Methylation utilized a single Polymerase V-like polymerase. Finally, we reveal that charophytic green algae encode a single CLSY/DRD1-type chromatin remodeling protein, further supporting the presence of a single specialized polymerase in charophytic green algae.
Subject(s)
DNA Methylation , DNA-Directed RNA Polymerases , Evolution, Molecular , DNA-Directed RNA Polymerases/genetics , DNA-Directed RNA Polymerases/metabolism , Phylogeny , Charophyceae/genetics , Charophyceae/enzymology , Plant Proteins/genetics , Plant Proteins/metabolism , Chlorophyta/genetics , Chlorophyta/enzymology , Protein Subunits/geneticsABSTRACT
Photostasis is the light-dependent maintenance of energy balance associated with cellular homeostasis in photoautotrophs. We review evidence that illustrates how photosynthetic adaptation in polar photoautrophs such as aquatic green algae, cyanobacteria, boreal conifers as well as terrestrial angiosperms exhibit an astonishing plasticity in structure and function of the photosynthetic apparatus. This plasticity contributes to the maintenance of photostasis, which is essential for the long-term survival in the seemingly inhospitable Antarctic and Arctic habitats. However, evidence indicates that polar photoautrophic species exhibit different functional solutions for the maintenance of photostasis. We suggest that this reflects, in part, the genetic diversity symbolized by inherent genetic redundancy characteristic of polar photoautotrophs which enhances their survival in a thermodynamically challenging environment.
Subject(s)
Adaptation, Physiological , Photosynthesis , Photosynthesis/physiology , Arctic Regions , Antarctic Regions , Cyanobacteria/physiology , Cyanobacteria/genetics , Chlorophyta/physiology , Chlorophyta/genetics , Ecosystem , Light , Magnoliopsida/physiology , Magnoliopsida/genetics , Tracheophyta/physiology , Tracheophyta/geneticsABSTRACT
Dunaliella salina is an innovative expression system due to its distinct advantages such as high salt tolerance, low susceptibility to contamination, and the absence of the cell wall. While nuclear transformation has been extensively studied, research on D. salina chloroplast transformation remains in the preliminary stages. In this study, we established an efficient chloroplast expression system for D. salina using Golden Gate assembly. We developed a D. salina toolkit comprising essential components such as chloroplast-specific promoters, terminators, homologous fragments, and various vectors. We confirmed its functionality by expressing the EGFP protein. Moreover, we detailed the methodology of the entire construction process. This expression system enables the specific targeting of foreign genes through simple homologous recombination, resulting in stable expression in chloroplasts. The toolkit achieved a relatively high transformation efficiency within a shorter experimental cycle. Consequently, the construction and utilization of this toolkit have the potential to enhance the efficiency of transgenic engineering in D. salina and advance the development of microalgal biofactories.
Subject(s)
Chloroplasts , Genetic Vectors , Green Fluorescent Proteins , Transformation, Genetic , Chloroplasts/genetics , Chloroplasts/metabolism , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Genetic Vectors/metabolism , Promoter Regions, Genetic , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Genetic Engineering/methods , Microalgae/genetics , Microalgae/metabolism , Chlorophyceae/genetics , Chlorophyceae/metabolism , Chlorophyta/genetics , Chlorophyta/metabolism , Homologous Recombination , Gene ExpressionABSTRACT
Ostreococcus spp. are unicellular organisms with one of the simplest cellular organizations. The sequencing of the genomes of different Ostreococcus species has reinforced this status since Ostreococcus tauri has one most compact nuclear genomes among eukaryotic organisms. Despite this, it has retained a number of genes, setting it apart from other organisms with similar small genomes. Ostreococcus spp. feature a substantial number of selenocysteine-containing proteins, which, due to their higher catalytic activity compared to their selenium-lacking counterparts, may require a reduced quantity of proteins. Notably, O. tauri encodes several ammonium transporter genes, that may provide it with a competitive edge for acquiring nitrogen (N). This characteristic makes it an intriguing model for studying the efficient use of N in eukaryotes. Under conditions of low N availability, O. tauri utilizes N from abundant proteins or amino acids, such as L-arginine, similar to higher plants. However, the presence of a nitric oxide synthase (L-arg substrate) sheds light on a new metabolic pathway for L-arg in algae. The metabolic adaptations of O. tauri to day and night cycles offer valuable insights into carbon and iron metabolic configuration. O. tauri has evolved novel strategies to optimize iron uptake, lacking the classic components of the iron absorption mechanism. Overall, the cellular and genetic characteristics of Ostreococcus contribute to its evolutionary success, making it an excellent model for studying the physiological and genetic aspects of how green algae have adapted to the marine environment. Furthermore, given its potential for lipid accumulation and its marine habitat, it may represent a promising avenue for third-generation biofuels.
Subject(s)
Chlorophyceae , Adaptation, Physiological , Chlorophyceae/cytology , Chlorophyceae/genetics , Chlorophyceae/metabolism , Chlorophyta/metabolism , Chlorophyta/genetics , Nitrogen/metabolism , Marine BiologyABSTRACT
Animal regeneration involves coordinated responses across cell types throughout the animal body. In endosymbiotic animals, whether and how symbionts react to host injury and how cellular responses are integrated across species remain unexplored. Here, we study the acoel Convolutriloba longifissura, which hosts symbiotic Tetraselmis sp. green algae and can regenerate entire bodies from tissue fragments. We show that animal injury causes a decline in the photosynthetic efficiency of the symbiotic algae, alongside two distinct, sequential waves of transcriptional responses in acoel and algal cells. The initial algal response is characterized by the upregulation of a cohort of photosynthesis-related genes, though photosynthesis is not necessary for regeneration. A conserved animal transcription factor, runt, is induced after injury and required for acoel regeneration. Knockdown of Cl-runt dampens transcriptional responses in both species and further reduces algal photosynthetic efficiency post-injury. Our results suggest that the holobiont functions as an integrated unit of biological organization by coordinating molecular networks across species through the runt-dependent animal regeneration program.
Subject(s)
Photosynthesis , Regeneration , Symbiosis , Animals , Regeneration/physiology , Chlorophyta/genetics , Transcription Factors/metabolism , Transcription Factors/geneticsABSTRACT
Mutualistic symbioses have contributed to major transitions in the evolution of life. Here, we investigate the evolutionary history and the molecular innovations at the origin of lichens, which are a symbiosis established between fungi and green algae or cyanobacteria. We de novo sequence the genomes or transcriptomes of 12 lichen algal symbiont (LAS) and closely related non-symbiotic algae (NSA) to improve the genomic coverage of Chlorophyte algae. We then perform ancestral state reconstruction and comparative phylogenomics. We identify at least three independent gains of the ability to engage in the lichen symbiosis, one in Trebouxiophyceae and two in Ulvophyceae, confirming the convergent evolution of the lichen symbioses. A carbohydrate-active enzyme from the glycoside hydrolase 8 (GH8) family was identified as a top candidate for the molecular-mechanism underlying lichen symbiosis in Trebouxiophyceae. This GH8 was acquired in lichenizing Trebouxiophyceae by horizontal gene transfer, concomitantly with the ability to associate with lichens fungal symbionts (LFS) and is able to degrade polysaccharides found in the cell wall of LFS. These findings indicate that a combination of gene family expansion and horizontal gene transfer provided the basis for lichenization to evolve in chlorophyte algae.
Subject(s)
Chlorophyta , Lichens , Phylogeny , Symbiosis , Lichens/genetics , Lichens/microbiology , Symbiosis/genetics , Chlorophyta/genetics , Gene Transfer, Horizontal , Evolution, Molecular , Biological Evolution , Transcriptome , Glycoside Hydrolases/genetics , Glycoside Hydrolases/metabolism , GenomicsABSTRACT
Dunaliella salina, a microalga that thrives under high-saline conditions, is notable for its high ß-carotene content and the absence of a polysaccharide cell wall. These unique characteristics render it a prime candidate as a cellular platform for astaxanthin production. In this study, our initial tests in an E. coli revealed that ß-ring-4-dehydrogenase (CBFD) and 4-hydroxy-ß-ring-4-dehydrogenase (HBFD) genes from Adonis aestivalis outperformed ß-carotene hydroxylase (BCH) and ß-carotene ketolase (BKT) from Haematococcus pluvialis counterparts by two-fold in terms of astaxanthin biosynthesis efficiency. Subsequently, we utilized electroporation to integrate either the BKT gene or the CBFD and HBFD genes into the genome of D. salina. In comparison to wild-type D. salina, strains transformed with BKT or CBFD and HBFD exhibited inhibited growth, underwent color changes to shades of red and yellow, and saw a nearly 50% decline in cell density. HPLC analysis confirmed astaxanthin synthesis in engineered D. salina strains, with CBFD + HBFD-D. salina yielding 134.88 ± 9.12 µg/g of dry cell weight (DCW), significantly higher than BKT-D. salina (83.58 ± 2.40 µg/g). This represents the largest amount of astaxanthin extracted from transgenic D. salina, as reported to date. These findings have significant implications, opening up new avenues for the development of specialized D. salina-based microcell factories for efficient astaxanthin production.
Subject(s)
Xanthophylls , Xanthophylls/metabolism , Chlorophyceae/metabolism , Chlorophyceae/genetics , Biosynthetic Pathways/genetics , Chlorophyta/metabolism , Chlorophyta/genetics , Escherichia coli/metabolism , Escherichia coli/genetics , Plant Proteins/metabolism , Plant Proteins/genetics , Mixed Function Oxygenases , OxygenasesABSTRACT
Steroid estrogens (SEs) have garnered global attention because of their potential hazards to human health and aquatic organisms at low concentrations (ng/L). The ecosystems of plateau freshwater lakes are fragile, the water lag time is long, and pollutants easily accumulate, making them more vulnerable to the impact of SEs. However, the knowledge of the impact of SEs on the growth and decomposition of phytoplankton communities in plateau lakes and the eutrophication process is limited. This study investigated the effects and mechanisms of SEs exposure on dominant algal communities and the expression of typical algal functional genes in Erhai Lake using indoor simulations and molecular biological methods. The results showed that phytoplankton were sensitive to 17ß-estradiol (E2ß) pollution, with a concentration of 50, and 100 ng/L E2ß exposure promoting the growth of cyanophyta and chlorophyta in the short term; this poses an ecological risk of inducing algal blooms. E2ß of 1000 ng/L exposure led to cross-effects of estrogenic effects and toxicity, with most phytoplankton being inhibited. However, small filamentous cyanobacteria and diatoms exhibited greater tolerance; Melosira sp. even exhibited "low inhibition, high promotion" behavior. Exposure to E2ß reduced the Shannon-Wiener diversity index (H'), Pielou index (J), and the number of dominant algal species (S) in phytoplankton communities, leading to instability in community succession. E2ß of 50 ng/L enhanced the expression levels of relevant functional genes, such as ftsH, psaB, atpB, and prx, related to Microcystis aeruginosa. E2ß of 50 ng/L and 5 mg/L can promote the transcription of Microcystis toxins (MC) related genes (mcyA), leading to more MC production by algal cells.
Subject(s)
Estradiol , Eutrophication , Lakes , Phytoplankton , Water Pollutants, Chemical , Phytoplankton/drug effects , Phytoplankton/genetics , Estradiol/toxicity , Water Pollutants, Chemical/toxicity , Diatoms/drug effects , Diatoms/genetics , Diatoms/metabolism , Diatoms/growth & development , Cyanobacteria/genetics , Cyanobacteria/metabolism , Cyanobacteria/drug effects , Chlorophyta/drug effects , Chlorophyta/genetics , Chlorophyta/growth & development , Chlorophyta/metabolismABSTRACT
BACKGROUND: Dark pigmented snow and glacier ice algae on glaciers and ice sheets contribute to accelerating melt. The biological controls on these algae, particularly the role of viruses, remain poorly understood. Giant viruses, classified under the nucleocytoplasmic large DNA viruses (NCLDV) supergroup (phylum Nucleocytoviricota), are diverse and globally distributed. NCLDVs are known to infect eukaryotic cells in marine and freshwater environments, providing a biological control on the algal population in these ecosystems. However, there is very limited information on the diversity and ecosystem function of NCLDVs in terrestrial icy habitats. RESULTS: In this study, we investigate for the first time giant viruses and their host connections on ice and snow habitats, such as cryoconite, dark ice, ice core, red and green snow, and genomic assemblies of five cultivated Chlorophyta snow algae. Giant virus marker genes were present in almost all samples; the highest abundances were recovered from red snow and the snow algae genomic assemblies, followed by green snow and dark ice. The variety of active algae and protists in these GrIS habitats containing NCLDV marker genes suggests that infection can occur on a range of eukaryotic hosts. Metagenomic data from red and green snow contained evidence of giant virus metagenome-assembled genomes from the orders Imitervirales, Asfuvirales, and Algavirales. CONCLUSION: Our study highlights NCLDV family signatures in snow and ice samples from the Greenland ice sheet. Giant virus metagenome-assembled genomes (GVMAGs) were found in red snow samples, and related NCLDV marker genes were identified for the first time in snow algal culture genomic assemblies; implying a relationship between the NCLDVs and snow algae. Metatranscriptomic viral genes also aligned with metagenomic sequences, suggesting that NCLDVs are an active component of the microbial community and are potential "top-down" controls of the eukaryotic algal and protistan members. This study reveals the unprecedented presence of a diverse community of NCLDVs in a variety of glacial habitats dominated by algae.
Subject(s)
Giant Viruses , Ice Cover , Ice Cover/virology , Greenland , Giant Viruses/genetics , Giant Viruses/classification , Giant Viruses/isolation & purification , Phylogeny , Ecosystem , Genome, Viral , Metagenomics , Chlorophyta/virology , Chlorophyta/genetics , Metagenome , SnowABSTRACT
The genomes of charophyte green algae, close relatives of land plants, typically do not show signs of developmental regulation by phytohormones. However, scattered reports of endogenous phytohormone production in these organisms exist. We performed a comprehensive analysis of multiple phytohormones in Viridiplantae, focusing mainly on charophytes. We show that auxin, salicylic acid, ethylene and tRNA-derived cytokinins including cis-zeatin are found ubiquitously in Viridiplantae. By contrast, land plants but not green algae contain the trans-zeatin type cytokinins as well as auxin and cytokinin conjugates. Charophytes occasionally produce jasmonates and abscisic acid, whereas the latter is detected consistently in land plants. Several phytohormones are excreted into the culture medium, including auxin by charophytes and cytokinins and salicylic acid by Viridiplantae in general. We note that the conservation of phytohormone biosynthesis and signaling pathways known from angiosperms does not match the capacity for phytohormone biosynthesis in Viridiplantae. Our phylogenetically guided analysis of established algal cultures provides an important insight into phytohormone biosynthesis and metabolism across Streptophyta.
Subject(s)
Cytokinins , Indoleacetic Acids , Phylogeny , Plant Growth Regulators , Plant Growth Regulators/metabolism , Indoleacetic Acids/metabolism , Cytokinins/metabolism , Viridiplantae/metabolism , Viridiplantae/genetics , Ethylenes/metabolism , Oxylipins/metabolism , Salicylic Acid/metabolism , Abscisic Acid/metabolism , Gene Expression Regulation, Plant , Cyclopentanes/metabolism , Biological Evolution , Chlorophyta/metabolism , Chlorophyta/genetics , Signal TransductionABSTRACT
Marine biogeochemical cycles are built on interactions between surface ocean microbes, particularly those connecting phytoplankton primary producers to heterotrophic bacteria. Details of these associations are not well understood, especially in the case of direct influences of bacteria on phytoplankton physiology. Here we catalogue how the presence of three marine bacteria (Ruegeria pomeroyi DSS-3, Stenotrophomonas sp. SKA14 and Polaribacter dokdonensis MED152) individually and uniquely impact gene expression of the picoeukaryotic alga Micromonas commoda RCC 299. We find a dramatic transcriptomic remodelling by M. commoda after 8 h in co-culture, followed by an increase in cell numbers by 56 h compared with the axenic cultures. Some aspects of the algal transcriptomic response are conserved across all three bacterial co-cultures, including an unexpected reduction in relative expression of photosynthesis and carbon fixation pathways. Expression differences restricted to a single bacterium are also observed, with the Flavobacteriia P. dokdonensis uniquely eliciting changes in relative expression of algal genes involved in biotin biosynthesis and the acquisition and assimilation of nitrogen. This study reveals that M. commoda has rapid and extensive responses to heterotrophic bacteria in ways that are generalizable, as well as in a taxon specific manner, with implications for the diversity of phytoplankton-bacteria interactions ongoing in the surface ocean.
Subject(s)
Photosynthesis , Transcriptome , Phytoplankton/genetics , Phytoplankton/metabolism , Chlorophyta/genetics , Chlorophyta/metabolism , Chlorophyta/microbiology , Bacteria/genetics , Bacteria/classification , Bacteria/metabolism , Heterotrophic Processes , Seawater/microbiologyABSTRACT
Green feather algae (Bryopsidales) undergo a unique life cycle in which a single cell repeatedly executes nuclear division without cytokinesis, resulting in the development of a thallus (>100 mm) with characteristic morphology called coenocyte. Bryopsis is a representative coenocytic alga that has exceptionally high regeneration ability: extruded cytoplasm aggregates rapidly in seawater, leading to the formation of protoplasts. However, the genetic basis of the unique cell biology of Bryopsis remains poorly understood. Here, we present a high-quality assembly and annotation of the nuclear genome of Bryopsis sp. (90.7 Mbp, 27 contigs, N50 = 6.7 Mbp, 14 034 protein-coding genes). Comparative genomic analyses indicate that the genes encoding BPL-1/Bryohealin, the aggregation-promoting lectin, are heavily duplicated in Bryopsis, whereas homologous genes are absent in other ulvophyceans, suggesting the basis of regeneration capability of Bryopsis. Bryopsis sp. possesses >30 kinesins but only a single myosin, which differs from other green algae that have multiple types of myosin genes. Consistent with this biased motor toolkit, we observed that the bidirectional motility of chloroplasts in the cytoplasm was dependent on microtubules but not actin in Bryopsis sp. Most genes required for cytokinesis in plants are present in Bryopsis, including those in the SNARE or kinesin superfamily. Nevertheless, a kinesin crucial for cytokinesis initiation in plants (NACK/Kinesin-7II) is hardly expressed in the coenocytic part of the thallus, possibly underlying the lack of cytokinesis in this portion. The present genome sequence lays the foundation for experimental biology in coenocytic macroalgae.
Subject(s)
Genome, Plant , Genome, Plant/genetics , Phylogeny , Chlorophyta/genetics , Chlorophyta/physiology , Regeneration/genetics , Bryopsida/genetics , Bryopsida/physiology , Bryopsida/cytology , Kinesins/genetics , Kinesins/metabolism , Myosins/genetics , Myosins/metabolismABSTRACT
Snow algae are a diverse group of extremophilic microeukaryotes found on melting polar and alpine snowfields. They play an important role in the microbial ecology of the cryosphere, and their propagation on snow and ice surfaces may in part accelerate climate-induced melting of these systems. High-quality snow algae genomes are needed for studies on their unique physiology, adaptive mechanisms, and genome evolution under multiple forms of stress, including cold temperatures and intense sunlight. Here, we assembled and annotated the genome of Limnomonas spitsbergensis, a cryophilic biciliate green alga originally isolated from melting snow on Svalbard, in the Arctic. The L. spitsbergensis genome assembly is based primarily on the use of PacBio long reads and secondly Illumina short reads, with an assembly size of 260.248â Mb in 124 contigs. A combination of 3 alternative annotation strategies was used including protein homology, RNA-seq evidence, and PacBio full-length transcript isoforms. The best merged set of annotations identified 18,277 protein-coding genes, which were 95.2% complete based on Benchmarking Universal Single-Copy Orthologs analysis. We also provide the annotated mitogenome, which is a relatively large 77.942â kb circular mapping sequence containing extensive repeats. The L. spitsbergensis genome will provide a new resource for research on snow algae adaptation, behavior, and natural selection in unique, low-temperature terrestrial environments that are under threat from climate change.
Subject(s)
Molecular Sequence Annotation , Snow , Arctic Regions , Snow/microbiology , Phylogeny , Chlorophyta/genetics , Genomics/methodsABSTRACT
Dunaliella bardawil is a marine unicellular green algal that produces large amounts of ß-carotene and is a model organism for studying the carotenoid synthesis pathway. However, there are still many mysteries about the enzymes of the D. bardawil lycopene synthesis pathway that have not been revealed. Here, we have identified a CruP-like lycopene isomerase, named DbLyISO, and successfully cloned its gene from D. bardawil. DbLyISO showed a high homology with CruPs. We constructed a 3D model of DbLyISO and performed molecular docking with lycopene, as well as molecular dynamics testing, to identify the functional characteristics of DbLyISO. Functional activity of DbLyISO was also performed by overexpressing gene in both E. coli and D. bardawil. Results revealed that DbLyISO acted at the C-5 and C-13 positions of lycopene, catalyzing its cis-trans isomerization to produce a more stable trans structure. These results provide new ideas for the development of a carotenoid series from engineered bacteria, algae, and plants.