Michael Melkonian

Oleaginous microalgae combine high lipid content with the ability to grow under diverse environmental conditions, the oleaginous species of family Scenedesmaceae (Sphaeropleales, Chlorophyta) having emerged as a promising source for biofuels, biostimulants, and bioremediation. This study provides long-read genome sequencing of 38 species/strains of Scenedesmaceae, including 16 chromosome-level genome assemblies, to explore the genomic underpinnings of the exceptional adaptive properties of their vegetative cells. Gene family analysis identifies 'core' and 'dispensible' orthogroups in the Scenedesmaceae and unveils orthogroup expansions and gains in four major metabolic processes that enhance survival of cells under stress: lipid metabolism, sulfur metabolism, resistance to oxidative stress, and heterotrophy. We discover orthogroup gains by horizontal gene transfer (HGT) from non-viridiplant donors in all four processes. Additionally, we identify ten diploid strains, which likely evolved by fusion of gametes and vegetative growth of the diploid cells. We hypothesize that HGT and diploidy fostered adaptive processes in the Scenedesmaceae.

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Biological soil crusts (biocrusts) cover the majority of the world’s dryland ground and are a significant component of the vegetation-free surface of the planet. They consist of an intimate association of microbial organisms, lichens, bryophytes and fungi. Biocrusts are severely endangered by anthropogenic disturbances despite their importance. The genus Bracteacoccus (Sphaeropleales, Chlorophyta) is a ubiquitous component of biocrusts from extreme environments. Here, we present the chromosome-level genome sequences of two Bracteacoccus species, B. bullatus and B. minor. Genome comparisons with other Archaeplastida identify genomic features that highlight the adaptation of these algae to abiotic stresses prevailing in such environments. These features include horizontal gene transfer events mainly from bacteria or fungi, gains and expansions of stress-related gene families, neofunctionalization of genes following gene duplications and genome structural variations. We also summarize transcriptional and metabolic responses of the lipid pathway of B. minor, based on multi-omics analyses, which is important for balancing the flexible conversion of polar membrane lipids and non-polar storage lipids to cope with various abiotic stresses. Under dehydration and high-temperature stress conditions B. minor differs considerably from other eukaryotic algae. Overall, these findings provide insights into the genetic basis of adaptation to abiotic stress in biocrust algae.

A total of 96 strains of Nostocales (Cyanobacteria) were established from the phyllosphere of the laurel forests in the Canary Islands (Spain) and the Azores (Portugal) using enrichment media lacking combined nitrogen. The strains were characterized by light microscopy and SSU rRNA gene comparisons. Morphologically, most strains belonged to two different morphotypes, termed “Nostoc-type” and “Tolypothrix-type”. Molecular phylogenetic analysis of 527 SSU rRNA gene sequences of cyanobacteria (95 sequences established during this study plus 392 sequences from Nostocales and 40 sequences from non-heterocyte-forming cyanobacteria retrieved from the databases) revealed that none of the SSU rRNA gene sequences from the phyllosphere of the laurel forests was identical to a database sequence. In addition, the genetic diversity of the isolated strains was high, with 42 different genotypes (44% of the sequences) recognized. Among the new genotypes were also terrestrial members of the genus Nodularia as well as members of the genus Brasilonema. It is concluded that heterocyte-forming cyanobacteria represent a component of the phyllosphere that is still largely undersampled in subtropical/tropical forests.

A total of 96 strains of Nostocales (Cyanobacteria) were established from the laurel forest phyllosphere in the Canary Islands (Spain) and Azores (Portugal) using enrichment media lacking combined nitrogen. The strains were characterized by light microscopy and SSU rRNA gene comparisons. Morphologically, most strains belonged to two different morphotypes, termed “Nostoc-type” and “Tolypothrix-type”. Molecular phylogenetic analysis of 527 SSU rRNA gene sequences of cyanobacteria (95 sequences established during this study plus 392 sequences from Nostocales and 40 sequences from non-heterocyte-forming cyanobacteria retrieved from the databases) revealed that none of the SSU rRNA gene sequences from the laurel forest phyllosphere was identical to a database sequence. In addition, the genetic diversity of the isolated strains was high with 42 different genotypes (44% of the sequences), recognized. Among the new genotypes were also terrestrial members of genus Nodularia as well as members of genus Brasilonema. It is concluded that heterocyte-forming cyanobacteria represent a still largely undersampled component of the phyllosphere in subtropical/tropical forests.

Porous substrate bioreactors (PSBRs) are a new technology to grow microalgae immobilized in a dense culture and solve some problems linked to suspended cultivation. During recent years, this technology has been used in laboratory and pilot setups in different fields of environmental biotechnology, such as wastewater treatment. The aim of this short review is to introduce the PSBR technology, summarize the results obtained in removing some pollutants from wastewater, provide an assessment of the potential of PSBRs for wastewater treatment, and the subsequent use of the algal biomass for other purposes.

Coccolithophores are well-known haptophytes that produce small calcium carbonate coccoliths, which in turn contribute to carbon sequestration in the marine environment. Despite their important ecological role, only two of eleven haptophyte plastid genomes are from coccolithophores, and those two belong to the order Isochrysidales. Here, we report the plastid genomes of two strains of Ochrosphaera neapolitana (Coccolithales) from Spain (CCAC 3688 B) and the USA (A15,280). The newly constructed plastid genomes are the largest in size (116,906 bp and 113,686 bp, respectively) among all the available haptophyte plastid genomes, primarily due to the increased intergenic regions. These two plastid genomes possess a conventional quadripartite structure with a long single copy and short single copy separated by two inverted ribosomal repeats. These two plastid genomes share 110 core genes, six rRNAs, and 29 tRNAs, but CCAC 3688 B has an additional CDS (ycf55) and one tRNA (trnL-UAG). Two large insertions at the intergenic regions (2 kb insertion between ycf35 and ycf45; 0.5 kb insertion in the middle of trnM and trnY) were detected in the strain CCAC 3688 B. We found the genes of light-independent protochlorophyllide oxidoreductase (chlB, chlN, and chlL), which convert protochlorophyllide to chlorophyllide during chlorophyll biosynthesis, in the plastid genomes of O. neapolitana as well as in other benthic Isochrysidales and Coccolithales species, putatively suggesting an evolutionary adaptation to benthic habitats.

Light-emitting diodes are increasingly used as artificial light sources in Haematococcus pluvialis cultivation due to the fact of their energy advantages. The immobilized cultivation of H. pluvialis in pilot-scale angled twin-layer porous substrate photobioreactors (TL-PSBRs) was initially performed with a 14/10 h light/dark cycle and showed relatively low biomass growth and astaxanthin accumulation. In this study, the illumination time with red and blue LEDs at a light intensity of 120 µmol photons m⁻² s⁻¹ was increased to 16–24 h per day. With a light/dark cycle of 22/2 h, the biomass productivity of the algae was 7.5 g m⁻² day⁻¹, 2.4 times higher than in the 14/10 h cycle. The percentage of astaxanthin in the dry biomass was 2%, and the total amount of astaxanthin was 1.7 g m⁻². Along with the increase in light duration, adding 10 or 20 mM NaHCO₃ to the BG11-H culture medium over ten days of cultivation in angled TL-PSBRs did not increase the total amount of astaxanthin compared with only CO₂ addition at a flow rate of 3.6 mg min⁻¹ to the culture medium. Adding NaHCO₃ with a 30–80 mM concentration inhibited algal growth and astaxanthin accumulation. However, adding 10–40 mM NaHCO₃ caused algal cells to accumulate astaxanthin at a high percentage in dry weight after the first four days in TL-PSBRs.

In the production of astaxanthin from Haematococcus pluvialis, the process of growing algal biomass in the vegetative green stage is an indispensable step in both suspended and immobilized cultivations. The green algal biomass is usually cultured in a suspension under a low light intensity. However, for astaxanthin accumulation, the microalgae need to be centrifuged and transferred to a new medium or culture system, a significant difficulty when upscaling astaxanthin production. In this research, a small-scale angled twin-layer porous substrate photobioreactor (TL-PSBR) was used to cultivate green stage biomass of H. pluvialis. Under low light intensities of 20–80 µmol photons m⁻²·s⁻¹, algae in the biofilm consisted exclusively of non-motile vegetative cells (green palmella cells) after ten days of culturing. The optimal initial biomass density was 6.5 g·m⁻², and the dry biomass productivity at a light intensity of 80 µmol photons m⁻²·s⁻¹ was 6.5 g·m⁻²·d⁻¹. The green stage biomass of H. pluvialis created in this small-scale angled TL-PSBR can be easily harvested and directly used as the source of material for the inoculation of a pilot-scale TL-PSBR for the production of astaxanthin.