Mechanisms protect airborne green microalgae during long distance dispersal.

Viable microalgae occur in the air. Whether they can survive the stresses such as UV, desiccation and freezing temperatures at high altitudes during long distance dispersal is rarely studied. If yes, what mechanisms confer the tolerance? Four freshwater airborne green microalgae were isolated from Dongsha Atoll in the South China Sea, classified as Scenedesmus sp. DSA1, Coelastrella sp. DSA2, Coelastrella sp. DSA3 and Desmodesmus sp. DSA6 based on their morphologies and ITS sequences. Their survival rates under UV stress were tightly correlated with their cell wall thickness. All the four airborne green microalgae survived the air-dry stress on benchtop followed by - 20 °C freeze-desiccation stress for 4 weeks, but not the two waterborne green microalgae Desmodesmus sp. F5 and Neodesmus sp. UTEX 2219-4 used as controls. Three of the four airborne microalgae survived the lyophilization treatment, excluding Desmodesmus sp. DSA6 and the two waterborne microalgae. The four airborne microalgae produced carotenoids under prolonged stress conditions, which might help detoxify the reactive oxygen species generated under environmental stresses and shield from the high-light stress in the air. Characterization of these airborne microalgae may help answer how the descendants of green algae survived on the land about 450 MYA.

VIPP2 interacts with VIPP1 and HSP22E/F at chloroplast membranes and modulates a retrograde signal for HSP22E/F gene expression.

VIPP proteins aid thylakoid biogenesis and membrane maintenance in cyanobacteria, algae, and plants. Some members of the Chlorophyceae contain two VIPP paralogs termed VIPP1 and VIPP2, which originate from an early gene duplication event during the evolution of green algae. VIPP2 is barely expressed under nonstress conditions but accumulates in cells exposed to high light intensities or H2 O2 , during recovery from heat stress, and in mutants with defective integration (alb3.1) or translocation (secA) of thylakoid membrane proteins. Recombinant VIPP2 forms rod-like structures in vitro and shows a strong affinity for phosphatidylinositol phosphate. Under stress conditions, >70% of VIPP2 is present in membrane fractions and localizes to chloroplast membranes. A vipp2 knock-out mutant displays no growth phenotypes and no defects in the biogenesis or repair of photosystem II. However, after exposure to high light intensities, the vipp2 mutant accumulates less HSP22E/F and more LHCSR3 protein and transcript. This suggests that VIPP2 modulates a retrograde signal for the expression of nuclear genes HSP22E/F and LHCSR3. Immunoprecipitation of VIPP2 from solubilized cells and membrane-enriched fractions revealed major interactions with VIPP1 and minor interactions with HSP22E/F. Our data support a distinct role of VIPP2 in sensing and coping with chloroplast membrane stress.

Regulation of Oxygenic Photosynthesis during Trophic Transitions in the Green Alga Chromochloris zofingiensis.

Light and nutrients are critical regulators of photosynthesis and metabolism in plants and algae. Many algae have the metabolic flexibility to grow photoautotrophically, heterotrophically, or mixotrophically. Here, we describe reversible Glc-dependent repression/activation of oxygenic photosynthesis in the unicellular green alga Chromochloris zofingiensis. We observed rapid and reversible changes in photosynthesis, in the photosynthetic apparatus, in thylakoid ultrastructure, and in energy stores including lipids and starch. Following Glc addition in the light, C. zofingiensis shuts off photosynthesis within days and accumulates large amounts of commercially relevant bioproducts, including triacylglycerols and the high-value nutraceutical ketocarotenoid astaxanthin, while increasing culture biomass. RNA sequencing reveals reversible changes in the transcriptome that form the basis of this metabolic regulation. Functional enrichment analyses show that Glc represses photosynthetic pathways while ketocarotenoid biosynthesis and heterotrophic carbon metabolism are upregulated. Because sugars play fundamental regulatory roles in gene expression, physiology, metabolism, and growth in both plants and animals, we have developed a simple algal model system to investigate conserved eukaryotic sugar responses as well as mechanisms of thylakoid breakdown and biogenesis in chloroplasts. Understanding regulation of photosynthesis and metabolism in algae could enable bioengineering to reroute metabolism toward beneficial bioproducts for energy, food, pharmaceuticals, and human health.

Taxonomic re-examination of "Chloromonas nivalis (Volvocales, Chlorophyceae) zygotes" from Japan and description of C. muramotoi sp. nov.

Recent molecular data has strongly suggested that field-collected cysts of snow algae that are morphologically identifiable as the zygotes of Chloromonas nivalis are composed of multiple species. Motile vegetative cells, however, have not been directly obtained from these cysts because of the difficulties involved in inducing their germination. Recently, our comparative molecular analyses, using both field-collected and cultured materials, demonstrated that one Japanese lineage of "C. nivalis zygotes" belongs to C. miwae. Herein, we examined another Japanese lineage of field-collected "C. nivalis zygotes" and a new strain originating from Japan. Our molecular data demonstrated that these two different life cycle stages are conspecific, and that they represent a new species that we herein describe as C. muramotoi sp. nov., based on the vegetative and asexual morphological characteristics of the strain. Multigene phylogenetic analyses showed that this new species was sister to C. miwae. Scanning electron microscopy demonstrated that the cysts of C. muramotoi are different from those of C. miwae, based on the arrangement of the flanges developing on the cell wall.

Three-dimensional ultrastructure and hyperspectral imaging of metabolite accumulation and dynamics in Haematococcus and Chlorella.

Phycology has developed alongside light and electron microscopy techniques. Since the 1950s, progress in the field has accelerated dramatically with the advent of electron microscopy. Transmission electron microscopes can only acquire imaging data on a 2D plane. Currently, many of the life sciences are seeking to obtain 3D images with electron microscopy for the accurate interpretation of subcellular dynamics. Three-dimensional reconstruction using serial sections is a method that can cover relatively large cells or tissues without requiring special equipment. Another challenge is monitoring secondary metabolites (such as lipids or carotenoids) in intact cells. This became feasible with hyperspectral cameras, which enable the acquisition of wide-range spectral information in living cells. Here, we review bioimaging studies on the intracellular dynamics of substances such as lipids, carotenoids and phosphorus using conventional to state-of-the-art microscopy techniques in the field of algal biorefining.

Subunit Asa3 ensures the attachment of the peripheral stalk to the membrane sector of the dimeric ATP synthase of Polytomella sp.

The mitochondrial ATP synthase of Polytomella exhibits a peripheral stalk and a dimerization domain built by the Asa subunits, unique to chlorophycean algae. The topology of these subunits has been extensively studied. Here we explored the interactions of subunit Asa3 using Far Western blotting and subcomplex reconstitution, and found it associates with Asa1 and Asa8. We also identified the novel interactions Asa1-Asa2 and Asa1-Asa7. In silico analyses of Asa3 revealed that it adopts a HEAT repeat-like structure that points to its location within the enzyme based on the available 3D-map of the algal ATP synthase. We suggest that subunit Asa3 is instrumental in securing the attachment of the peripheral stalk to the membrane sector, thus stabilizing the dimeric mitochondrial ATP synthase.