Thiamine metabolism genes in diatoms are not regulated by thiamine despite the presence of predicted riboswitches

Thiamine pyrophosphate (TPP), an essential co-factor for all species, is biosynthesised through a metabolically expensive pathway regulated by TPP riboswitches in bacteria, fungi, plants and green algae. Diatoms are microalgae responsible for approximately 20% of global primary production. They have been predicted to contain TPP aptamers in the 3'UTR of some thiamine metabolism-related genes, but little is known about their function and regulation. We used bioinformatics, antimetabolite growth assays, RT-qPCR, targeted mutagenesis and reporter constructs to test whether the predicted TPP riboswitches respond to thiamine supplementation in diatoms. Gene editing was used to investigate the functions of the genes with associated TPP riboswitches in Phaeodactylum tricornutum. We found that thiamine-related genes with putative TPP aptamers are not responsive to thiamine or its precursor 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP), and the targeted mutation of the TPP aptamer in the HMP-P synthase (THIC) does not deregulate thiamine biosynthesis in P. tricornutum. Through genome editing we established that PtSSSP is necessary for thiamine uptake and that PtTHIC is essential for thiamine biosynthesis. Our results highlight the importance of experimentally testing bioinformatic aptamer predictions and provide new insights into the thiamine metabolism shaping the structure of marine microbial communities with global biogeochemical importance.

Media engineering in marine diatom Phaeodactylum tricornutum employing cost-effective substrates for sustainable production of high value renewables

Phaeodactylum tricornutum is a marine diatom, and well-studied model of unicellular microalga. This diatom contains a wide range of high-value renewables (HVRs) with high commercial relevance owing to their importance in human nutrition and health. In this study, we screened P. tricornutum for biomass, eicosapentaenoic acid (EPA) and fucoxanthin production under photoautotrophic and mixotrophic condition with various substrate combinations. Results highlights that culture supplemented with glycerol and urea lead to enhanced biomass, biochemical and HVR production. Further continuous feeding of urea in glycerol supplemented medium results in an increase in biomass yield (0.77 g L-1) by ~ 2-fold. Additionally, continuous feeding of urea channelizes the carbon flux towards biosynthesis of fatty acids increasing FAME content by ~2-fold as compared to the control conditions. Overall EPA and fucoxanthin production was 27 mg L-1 and 11 mg L-1 (~2 & 4 fold) in urea fed cultures respectively. Present study demonstrates efficient valorization of cost-effective substrates such as glycerol and urea for the production of high-value renewables in P. tricornutum.

Toxicity of environmentally important micropollutants on three trophic levels

Micropollution is a serious environmental problem caused by continuous entry of trace quantities of toxic chemical substances into the aquatic environment. In the present study, three trophic levels of the aquatic ecosystems were used to evaluate the acute toxicities of environmentally important micropollutants including heavy metals, pesticides and drugs. There is a scarcity of information on toxicity of the studied substances on marine water algae. Among studied micropollutants, the most toxic chemical to Daphnia magna and Danio rerio was found to be 1-Chloro-2,4 dinitrobenzene with EC50 of 0.002 and 4.2 mg/L, respectively. Although this compound was also toxic to marine algae, Phaeodactylum tricornutum, arsenic showed the highest toxicity to the algae with EC50 of 2.4 mg/L. As compared to other organisms, D. magna was found to have higher sensitivity to all of the tested micropollutants.

FUNCTIONAL STATE OF THE MARINE MICROALGAE CULTURES AS AN INDICATOR OF THE WATER POLLUTION LEVEL OF THE SEVASTOPOL BAY

The possibility of using cultures of Black Sea planktonic microalgae of different taxonomic affiliation to assess the quality of the aquatic environment based on the assessment of their functional state is shown. The research was carried out on the diatom Phaeodactylum tricornutum, three dinoflagellates species Prorocentrum cordatum, Prorocentrum pusillum and Gyrodinium fissum, as well as the coccolithopho-ride Emiliania huxleyi. It was found that the waters of the Sevastopol Bay in the period from May to August 2021 in 50% of cases had a weak inhibitory effect on the growth of the tested species. In other cases, algae growth was either stimulated or the effect of pollution on cultures was not revealed. The need for using several types of algae in carrying out complex work on water biotesting of the coastal are-as of the Black Sea is highlighted.

A Multi-Species Investigation of Sponges’ Filtering Activity towards Marine Microalgae

Chronic discharge of surplus organic matter is a typical side effect of fish aquaculture, occasionally leading to coastal eutrophication and excessive phytoplankton growth. Owing to their innate filter-feeding capacity, marine sponges could mitigate environmental impact under integrated multitrophic aquaculture (IMTA) scenarios. Herein, we investigated the clearance capacity of four ubiquitous Mediterranean sponges (Agelas oroides, Axinella cannabina, Chondrosia reniformis and Sarcotragus foetidus) against three microalgal substrates with different size/motility characteristics: the nanophytoplankton Nannochloropsis sp. (~3.2 μm, nonmotile) and Isochrysis sp. (~3.8 μm, motile), as well as the diatom Phaeodactylum tricornutum (~21.7 μm, nonmotile). In vitro cleaning experiments were conducted using sponge explants in 1 L of natural seawater and applying different microalgal cell concentrations under light/dark conditions. The investigated sponges exhibited a wide range of retention efficiencies for the different phytoplankton cells, with the lowest average values found for A. cannabina (37%) and the highest for A. oroides (70%). The latter could filter up to 14.1 mL seawater per hour and gram of sponge wet weight, by retaining 100% of Isochrysis at a density of 105 cells mL−1, under darkness. Our results highlight differences in filtering capacity among sponge species and preferences for microalgal substrates with distinct size and motility traits.