Journal of Phycology

Marine non-cyanobacterial diazotrophs (NCDs) are recognized as globally distributed, however, few representatives have been isolated in pure cultures. As a result, understanding the physiology, growth rate, substrate preference and dinitrogen (N2) fixation capabilities proves difficult. Thalassolituus haligoni. sp. nov., BB40 was isolated from a fjord-like inlet within Kjipuktuk (Halifax), Nova Scotia. The fully sequenced genome displayed all necessary genes required for N2 fixation, and various carbon uptake pathways. The gram-negative flagellated rod shape bacterium displayed significantly higher growth rates in medium amended with nitrate (NO3-) or ammonia (NH3), compared to dissolved N2, as the sole nitrogen source. Biological N2 fixation rates were detectable across all conditions, measuring a range from 9.34 x 10-6 to 1.4 x 10-1 fmol N cell-1 day-1. Growth of the isolate was successful between 4 {degrees}C up to 35 {degrees}C, with a Topt of 20 {degrees}C for N2, and between 27 - 30 {degrees}C for fixed nitrogen (NO3- and NH3). The closest relatives to T. haligoni, were found to be the uncultured Arc-gamma-03 (99% average nucleotide identity (ANI)) and Oceanobacter antarcticus (81% ANI). T. haligoni also displays versatile capabilities for growth on various carbon, and nitrogen sources, and antibiotics. Collectively this study provides an in-depth physiological assessment of an Oceanospirillales diazotrophic species which we presently have limited knowledge of.

Amino acid catabolism is a vital metabolic process in bacteria, providing energy, carbon and potentially nitrogen as resources, and affecting global cycles of these elements. The ability of a bacterium to catabolize an amino acid is often inferred from the presence of the relevant catabolic pathways in its genome, yet the "gene=function" inference is not straightforward. Here, we use growth assays in 96 well plates on individual amino acids and their combinations to directly measure the ability of a model marine bacterium, Alteromonas macleodii ATCC 27126, to utilize these resources for growth. With the exception of aspartate and glutamate, which did not support growth in any of our experiments, ATCC 27126 grew on all other amino acids. However, the probability of growth, together with growth yield and rate, differed depending on the entry point of the catabolic pathway to central carbon metabolism, with robust growth occurring only on amino acids catabolized into pyruvate or acetyl CoA. Growth on combinations of two amino acids revealed reproducible patterns, the clearest being inhibition of growth on other amino acids by asparagine, aspartate and their degradation product, oxaloacetate. Finally, growth was different in test tubes compared with 96 well plates. Our results reveal hidden complexity in amino acid utilization and suggest a "TCA-centric" viewpoint for amino acid utilization, perhaps reflecting the high metabolic flexibility of pyruvate and specific regulatory aspects of the TCA cycle in Alteromonas.

Recently, an inventory of genes in phytoplankton was conducted through expeditions such as TARA Oceans. Approximately 1.5 million genes were identified, of which at least three-quarters have unknown function. Presently, a several research programmes are engaged in the sequencing of marine biodiversity, resulting in a rapid expansion of genomic databases. Access to the genomic sequences of these organisms will soon be readily accessible to the scientific community. Although analysing this data is promising, the characterization of genes or genomes, on the other hand, is progressing very slowly and remains a major challenge for scientists. The aim of this study was to use GWAS approaches to decipher genomic loci without a priori assumptions. The microalga Tisochrysis lutea was selected as a case study due to its economic importance and the extensive knowledge accumulated over the years. Particular attention was paid to pigment and lipid metabolism due to their high commercial value. To implement the GWAS approach, a collection of algal lineages was established (100 lineages) from available polyclonal strains (15 strains). This collection was then phenotyped under two different culture conditions. Of the 31 phenotypic traits investigated, 18 met the requirements for GWAS analysis. Concurrently, each algal lineage was genotyped by whole genome sequencing to inventory all genetic polymorphisms. A mixed model was applied, revealing 13 significant associations between phenotypic traits and alleles. These associations highlight previously unsuspected genomic loci that play a major role in pigment or lipid content. Genes identified at these loci may have a direct or indirect role in these metabolic pathways. Nevertheless, elucidating the molecular mechanisms of the associated genes remains limited without the implementation of functional approaches. Despite the complexity of the process, we conclude that the GWAS approach was effective for deciphering phytoplankton genomes, particularly for quantitative traits of interest. Ideally, this approach should be combined with other functional methods to progressively decode marine genomes.

The size structure of phytoplankton communities plays a key role in the fate of carbon fixed by photosynthesis. Whether phytoplankton cells sink, enter the microbial loop, or are consumed by larger organisms is generally determined by their size. Grazing has been advanced as a factor determining size structure, but sources of mortality other than grazing, such as viruses also are recognized to be important. Based on the observation that cell size and genome size are related in phytoplankton, we hypothesize that viruses can also play a role in shaping the size structure of the phytoplankton community. Because cell size is related to genome size, we suggest that phytoplankton species with larger genomes will have a more developed immune system to defend against viral infection. As a first step to test this hypothesis, we screened the published transcriptomes of 125 phytoplankton species for expressed viral and immune-response related genes. We found a significant negative correlation between host-cell size and viral-gene diversity, and a positive correlation between host-cell size and the number of immune-response related genes. Our hypothesis supported by preliminary findings opens new pathways to explore whether we should consider viruses as an additional evolutionary driver for larger phytoplankton size, along with grazing and nutrients.

O_LIPhytoplankton are the major primary producers in the Southern Ocean, participate in the global carbon cycle, nutrient cycles, and are at the base of the food-web. These polar ecosystems are unique in their extended periods of darkness in the winter. C_LIO_LIProlonged darkness has the potential to exert selection that affects the composition of diatom communities if there is differential survival of diatoms in the dark, variation in population growth rates in subsequent light periods, or both. C_LIO_LIWe tested whether prolonged darkness has the potential to exert within-species selection on a model polar diatom species by exposing 5 strains of the polar diatom Porosira glacialis to prolonged darkness at two different temperatures in the laboratory. We measured population survival in the dark, growth rate upon re-illumination, and between strain variability in these traits. C_LIO_LIWe found a pronounced decline in survival and growth rate with time spent in the dark, as well as important intraspecific variation in these. C_LIO_LIHigher temperature exacerbated declines in growth and survival. C_LIO_LIOur results show that the darkness of polar night can exert selection within diatom species, with implications for phytoplankton community composition and subsequent impacts on Southern Ocean biogeochemical cycles. C_LI

Tropical coral reef ecosystems worldwide are being impacted by combined pressures of climate change and human activities that introduce large quantities of nutrients and sediments into coastal areas. In this context, phytoplankton represent a critical link between dissolved inorganic nutrients and coral reef food webs, yet their role in these ecosystems remains understudied. We investigated ecological responses of the summer phytoplankton community of Shiraho Reef (Ishigaki Island, Okinawa, Japan) to nutrient enrichment using field-based microcosm experiments under natural light and temperature conditions in September 2022 and 2023. Treatments included single and combined additions of nitrogen, phosphorus, and silicon. Chlorophyll a (Chl a) concentrations increased after three days under combined nutrient conditions, whereas single-nutrient additions produced limited responses, indicating a strong co-limitation by nitrogen and phosphorus in the reef. Analysis of size-fractionated Chl a revealed shifts from picophytoplankton that typically dominate tropical oligotrophic ecosystems toward larger groups supported by enhanced nutrient availability. Our results show short-term impacts of nutrient enrichment events on phytoplankton size structure and biogeochemical cycling in coral reefs, and highlight the importance of pelagic processes in coral reef carbon dynamics under nutrient-enrichment.

Microphytobenthos (MPB) contributes significantly to the marine primary production in estuarine ecosystems. MPB is mainly composed of benthic motile diatoms inhabiting intertidal and shallow subtidal sediments. Unlike intertidal small-sized diatom models, subtidal ([≥] 10 m depth) MPB and large-sized (>100 {micro}m) species, have comparatively received much less attention, especially as regards to their photosynthetic productivity. Yet, the subtidal light environment shows unusual (very) low intensities and a green-blue light spectrum at high tide. The present study investigates the light-dependent green-blue responses of the subtidal giant diatom Pleurosigma strigosum, combining in situ monitoring with laboratory experiments. In both MPB and P. strigosum, we documented a strong photophysiological plasticity, and a striking alignment with the photophysiology of (very) low light-adapted polar diatoms. Altogether, our results highlight the nature of subtidal photoadaptation: a very low green-blue light sensitive response, which explains the early blooming of P. strigosum at the very beginning of spring and underpins the ecological success of MPB in colonizing coastal subtidal sediments. The general coherence between subtidal P. strigosum and MPB light-responses offers a unique model species and growth form to further decipher the specific light-driven metabolism of subtidal MPB under precisely controlled environmental conditions.

Ostreobium, a siphonous green alga capable of living inside of calcium carbonate substrates, including the skeletons of reef-building corals. This study investigates spectral niche preferences and physiological strategies of Ostreobium using community-wide experiments. We exposed natural Ostreobium communities from Porites lutea collected across shallow, mid, and deeper-water sites to three light conditions: far-red, blue, and white light, simulating healthy shallow-water corals, deeper water conditions, and bleached coral skeletons respectively. Using 16S rRNA metabarcoding and chlorophyll analysis, we assessed community changes and physiological responses over 16 weeks. We show significant variation in spectral preferences among Ostreobium OTUs, with clear evidence for both generalist and specialist strategies. Chlorophyll analysis showed photoacclimation responses through changes in pigment compositions. Our work shows that the spectral architecture of the reef plays a role in structuring Ostreobium communities, but the many mismatches between spectral preferences of OTUs and their observed presence in nature, suggests that inter-species competition is likely to be an even stronger contributor to community structure across the reefs microhabitats. We show that physiological heterogeneity within Ostreobium is strongly phylogenetically structured, and our results clearly highlight the importance of considering OTU-level differences when predicting community responses to environmental disturbances such as coral bleaching. While generalist OTUs dominate natural communities, these do poorly in incubations, and we hypothesise that white light specialists may become key players during coral bleaching events. Our work is a substantial advance in our understanding of Ostreobium ecology and provides a framework for interpreting future environmental sequencing data, offering insights into the functional roles of the different OTUs.

European tidal flats that host non-native Magallana gigas reefs contribute to several ecosystem functions. Among others, they provide a habitat for a large variety of associated fauna. However, we often lack detailed information about any trophic interactions of the associated macrozoobenthos species with the oysters, and about their role in the carbon and nutrient cycle. Therefore, we performed ex-situ pulse-chase tracer experiments in the Eastern Scheldt (Southwest Dutch Delta, Netherlands) in summer and autumn 2020, where we fed M. gigas and their associated fauna 13C- and 15N-enriched bacterioplankton while the macrozoobenthos was incubated in water containing deuterium oxide (2H2O; enrichment: 1 - 2.5%). The aim was (1) to assess differences in short-term (<12h) processing of bacterioplankton in summer and autumn, and (2) to study differences in 2H incorporation - a proxy for metabolic activity - of M. gigas and its associated fauna in summer and autumn. In summer, all macrozoobenthos species combined consumed significantly less bacterioplankton-derived 13C and 15N than in autumn, while all macrozoobenthos species combined incorporated comparable amounts of 2H into their tissue in both seasons. Most bacterioplankton-derived 13C and 15N was taken up by sponges (Halichondria panicea, Hymeniacidon perlevis), crabs (Carcinus maenas, Eriocheir sinensis, Rhithropanopeus harrisii), and limpets (Crepidula fornicata). Most 2H was taken up by crabs (C. maenas, E. sinensis), sponges (H. perlevis), and snails (Littorina littorea), implying that these species were the most metabolically active ones. Overall, the metabolic activity was linked to feeding activity in summer 2020, whereas in autumn 2020, the link was weaker and the most metabolically active species were not necessarily the species that had incorporated most 13C and/or 15N.

Nitrogen is an important element for all living organisms. Photoautotrophic organisms need to assimilate nitrogen from the environment, therefore changes in nitrogen availability have a strong influence on their growth and metabolism. Many microalgae have been known to contain crystalline inclusions, and recently, it has been shown that many of these consist of purines like guanine and thus must be linked to the cellular nitrogen metabolisms. The alveolate alga Chromera velia contains such guanine crystals, and during its lifecycle, the alga is thought to be subjected to strong changes in external nitrogen availability. Here, we investigated the formation or decline of crystalline guanine in dependence of the availability of inorganic nitrogen in the growth medium. Cells were examined using polarised light microscopy, Raman micro-spectroscopy, chromatography (HPLC), transmission and scanning electron microscopy. The cellular guanine crystal content decreased during nitrogen starvation and increased upon transfer of the cells back to standard growth medium containing nitrate. Raman micro-spectroscopy showed that the crystals were composed of anhydrous guanine in beta-polytype. They appear in unspecific positions throughout the cell, and staining with the green dye Lysotracker DND-26 suggests that they are within vacuoles. Stacks of crystals could be observed in cells via freeze fracture and freeze etching electron microscopy, which unambiguously showed a membrane around the crystal aggregates, in a similar arrangement as has been shown for guanine storage vacuoles (GSV) in Chlamydomonas reinhardtii. We developed a method to isolate the guanine crystals from whole cells, and were able to obtain crystals which retained their flat, plate-like structure, matching the electron microscopic observations from whole cells. The isolated crystals were shown to consist of nitrogen rich compounds via energy-dispersive X-ray (EDX) analysis, and Raman micro-spectroscopy confirmed that they consist of guanine.

Environmental DNA (eDNA) metabarcoding is a key tool in biodiversity monitoring due to its high-throughput, non-destructive nature. While short-read (SR) sequencing platforms such as Illumina Miseq have been routinely used in environmental monitoring, their limited read lengths (less than 600 bp) constrain the depth of taxonomic assignment, particularly for complex microbial eukaryotes like protists. Conversely, long-read (LR) sequencing technologies like Oxford Nanopore Technologies (ONT) offer promising alternatives but remain underutilized for studying protist communities. We conducted a comparative study of SR versus LR metabarcoding of protist communities along a coastal-offshore gradient in the Belgian part of the North Sea. Using amplicons targeting the V4 region (SR; 577 bp) and the V4-V5 region (LR; 745 bp) of the 18S rRNA gene, we compared diversity patterns, taxonomic assignment, and community composition between approaches. We observed general congruence in community composition at higher taxonomic levels, but under the applied workflows, LR metabarcoding yielded a greater depth of taxonomic annotation at lower taxonomic ranks. Notably, dinoflagellates were less overrepresented in LR data, and a unique detection of potential nuisance taxa (e.g., Bellerochea), and ecologically important genera such as haptophytes (e.g., Gephyrocapsa) was achieved. These results highlight the potential of LR metabarcoding to complement SR approaches by providing increased taxonomic annotation depth and ecological insights. Although both methods targeted only partial regions of the 18S rRNA gene, LR metabarcoding yielded a greater depth of taxonomic assignment under the applied workflows. As next-generation sequencing technologies continue to evolve, our research provides valuable insights for selecting optimal strategies in routine plankton monitoring and biodiversity assessment programs.

Three novel bioluminescent bacterial strains, VNB-15T, VNB-16 and SChm4, were isolated from water of the Black Sea (Russia) and intestines of the Black Sea horse mackerel. Cells of the isolated strains are motile Gram negative slightly curved rods with single polar flagellum. The temperature range for growth was 10-35{degrees}C, the optimum being 20-25{degrees}C. The pH range for growth was 6.0-9.0, the optimum being 7.0-8.0. The bacteria were able to grow in the presence of 0.5 to 5.0% NaCl (w/v), the optimum being 1.0-4.0% (w/v). Phylogenetic analysis based on comparison of 16S rRNA sequences shows these strains to have kinship with the species Vibrio jasicida, Vibrio hyugaensis, Vibrio alginolyticus, Vibrio campbelli, Vibrio rotiferianus, Vibrio harveyi and Vibrio owensii with sequence similarity from 99.6 to 98.0%. Phylogenetic analysis based on comparison of the sequences of genes gyrB, recA, pyrH, gapA, rpoA, mreB, ftsZ, topA shows that the strains VNB-15T, VNB-16 and SChm4 to form a cluster within the V. harveyi clade and belong to a new species of the Vibrio genus. Comparison of the complete genomic sequence of VNB-15T with typical strains of nearby species also indicates that VNB-15T belongs to a separate species (maximum similarity 98% with V. hyugaensis and 96% with V. jasicida). VNB-15T differs from closely related species by its ability to utilize glucose, mannitol, inositol, sorbitol, rhamnose and sucrose, and to form lysine decarboxylase, ornithine decarboxylase, lipase, acid phosphatase, -glucosidase, {beta}-glucosidase and N-acetyl-{beta}-D-glucosaminidase enzymes. Based on phylogenetic analysis and phenotypic characteristics, Vibrio aquamarinus sp. nov. is proposed. The type strain is VNB-15T (= VKPM B-11245T = DSM 26054 T). RepositoriesThe GenBank accession numbers for the gapA, 16S rRNA, gyrB, pyrH, rpoA, recA, mreB, ftsZ, topA genes sequences of strain VNB-15T are JQ319116-JQ319121, KX242381, KX 242384, KX242387, respectively. The GenBank accession numbers for the gapA, gyrB, pyrH, recA, rpoA,16S rRNA, mreB, ftsZ, topA genes sequences of strain VNB-16 are KP221561-KP221566, KX242382, KX 242385, KX242388 respectively. The GenBank accession numbers for the 16S rRNA, gapA gyrB, rpoA, recA, pyrH, mreB, ftsZ, topA genes sequences of strain SChm4 are KX242375-KX242380, KX242383, KX 242386, KX242389, respectively. The GenBank/EMBL/DDBJ accession numbers for the housekeeping gene sequences used in this study are detailed in supplementary Table S1, Figures S1-S8. The genome of Vibrio aquamarinus sp. nov. VNB-15T, comprising two chromosomes and a plasmid, has been assembled and deposited in the NCBI database under the submission number SUB14585067

Many commercially exploited fish stocks have declined over the last few decades. It is therefore essential to identify natural populations and understand local adaptation for sustainable management. Salinity is a key environmental factor shaping local adaptation, and adaptive trait divergence often occurs at the egg and larval stages. The strong salinity gradient in the brackish Baltic Sea has driven rapid adaptation in multiple taxa. The Eastern Baltic cod (Gadus morhua) has adapted to low salinity with buoyant and tolerant eggs and larvae, but the stock has declined both in abundance and geographical range during the last decades. The main reproduction area of this stock is in the Bornholm Basin (ICES subdivision (SD) 25) in the southern Baltic Proper. Cod in this area, however, exhibit stunted growth and small body sizes. In contrast, large and healthy cod in reproductive condition have been observed in the [A]land Sea in the northern Baltic Proper (SD 29), raising the question of whether these fish represent a locally adapted population capable of successful reproduction in the lower salinities (5-10 psu in the northern Baltic Proper (SD 27, 29 and 32). Here, we experimentally assessed egg and yolk-sac larvae survival across salinities, egg size, egg and larval neutral buoyancy and egg survival on sediment, to test whether northern ([A]land) cod show adaptation to low salinity at early life stages as compared to southern cod. Mortality of larvae increased with decreasing salinity in cod from both areas, with the lowest survival at 7 psu. At 9 psu, more than 50% of northern cod larvae survived, suggesting that development could occur in SD29. Egg size and buoyancy were similar between northern and southern cod, and eggs and larvae were negatively buoyant, sinking under local salinity conditions. Nevertheless, the eggs survived and hatched well on sediment, indicating potential for demersal spawning. Our findings show no strong evidence of adaptive divergence to lower salinity in northern cod; however, their ability to tolerate sediment contact at early life stages suggests that Eastern Baltic cod may reproduce outside their historical spawning grounds.

Biologists aim to predict where species will survive and thrive as the planet warms. To do so, we often rely on data-hungry species distribution models (SDMs) that use associations between species occurrences and environmental predictors to capture the realised niche. An alternative basis for predictions is to experimentally quantify the effect of environmental drivers on performance, which captures the fundamental niche. We presently do not know which of these approaches represents a better path towards accurate forecasts. SDMs may depend too strongly on present-day environmental covariation, which will change in the future. In contrast, a major shortcoming of experiments is that they ignore most environmental drivers to focus on one or two. Quantifying how well fundamental and realised niches agree today would help establish how useful both SDMs and experiments are likely to be. We therefore compared both niches in 39 relatively common marine phytoplankton species. The temperature-dependence of population growth rate was characterised with a thermal performance curve model applied to lab experimental data, and the temperature-dependence of species occurrence probability estimated with SDMs applied to a global compilation of marine presence records. We found a fairly strong, near 1:1 relationship between measures of thermal niche centre: the median growth temperature in the lab and the median occurrence temperature in the field (R2 = 0.49). We also found a modest positive relationship between measures of thermal niche width, the growth niche width and the occurrence niche width (R2 = 0.24). This agreement should increase our confidence in environmental preferences inferred with SDMs. It also suggests that simple experiments can reliably constrain species ranges and help forecast range shifts. This has important implications for forecasting community composition and ecosystem processes, as we ought to be able to predict range shifts in biogeochemically-important taxa such as diatoms and nitrogen-fixing cyanobacteria.

BackgroundRed macroalgae (Rhodophyta) are ecologically and economically important marine primary producers, yet genomic resources for most species remain scarce. Delisea pulchra, a temperate red alga known for its halogenated furanone-based chemical defenses, serves as a model for studying algal-microbe interactions, antifouling mechanisms, and disease dynamics. ResultsHere we present a high-quality genome assembly of this species. The nuclear genome comprises 134 Mbp across 271 contigs with an N50 of 1.47 Mbp and encodes 13,387 predicted protein-coding genes. Comparative genomics with other red algae revealed expansions in gene families involved in DNA methylation, and oxidative stress responses, including glutathione S-transferases and superoxide dismutases. Analysis of glycosyltransferases, sulfatases, and sulfurylases implicated in galactan biosynthesis suggests D. pulchra possesses a complex and potentially novel extracellular matrix. We also identified several vanadium haloperoxidases (vHPOs), heme-dependent haloperoxidases (hHPOs), and two type III polyketide synthase (PKS) genes unique to the D. pulchra, which together represent promising candidate genes for bromofuranone production. ConclusionThe D. pulchra genome provides a foundation for molecular investigations into defense, signaling, and host-microbe interactions. It has been deposited at the European Nucleotide Archive under accession number PRJEB101077. All datasets, annotations, and interactive tools for exploring the genome are also available through the Rhodoexplorer portal at https://rhodoexplorer.sb-roscoff.fr.

Mesopredators contribute to food web stability and as such, understanding their trophic ecology can help to predict potential consequences of ongoing ecosystem modification. Here, multi-tissue carbon and nitrogen stable isotope analysis ({delta}13C and {delta}15N) and biochemical blood parameters ({beta}-hydroxybutyrate, glucose, lactate, and osmolality) were used to assess sex, size, spatial and seasonal differences in trophic ecology and condition of southern stingrays, Hypanus americanus, in Bimini, The Bahamas. Stingrays exhibited a dietary preference for molluscs and annelids, with an ontogenetic shift towards lower {delta}13C with increasing body size indicating a shift towards more mangrove associated prey. Nitrogen isotope values showed minimal seasonal changes, but higher {delta}15N values in males indicated foraging at a higher trophic level than females. Blood {beta}-hydroxybutyrate concentrations and osmolality revealed a similar energetic state and condition between sex, size, location and season. Our results advance our understanding of the seasonal trophic ecology of a benthic, marine mesopredator and identify the southern stingray as an important trophic link in seagrass and mangrove habitats in Bimini.

Mixotrophy is emerging as a default nutritional strategy in phytoplankton but research seems so far isolated and mostly focussing on single phytoplankton groups or strains. Here we combined data from 24 oceanic and 22 freshwater strains - as well as results from other studies - to analyze phytoplanktons ability to utilize dissolved organic compounds and highlight potential influencing factors. The results emphasize that mixotrophy is ubiquitous in phytoplankton across functional groups and taxa isolated from various habitats, and not strictly dependent on light or nutrient deficiencies. Several factors such as taxonomic affiliation, temperature and growth phase can affect mixotrophic behavior but no consistent patterns have emerged regarding their effects. Hence, mixotrophic traits remain so far unpredictable. There is some indication that the strains origin - potentially through adaptation to habitat DOM availability - might predetermine phytoplanktons mixotrophic skills. For example, freshwater strains used overall more compounds than oceanic strains in our study and Ostreococcus exhibited a different use pattern depending on its origin. Nevertheless, many aspects of mixotrophy in phytoplankton - e.g. metabolic pathways - remain cryptic. By summarizing available knowledge and knowledge gaps, the present synthesis provides a guideline for upcoming research further exploring mixotrophy. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/703725v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@12ac311org.highwire.dtl.DTLVardef@6c98deorg.highwire.dtl.DTLVardef@1a837ddorg.highwire.dtl.DTLVardef@eb9b53_HPS_FORMAT_FIGEXP M_FIG C_FIG

The raphidophyte Chattonella marina is a harmful algal bloom (HAB) species known for its distinct diurnal vertical migration (DVM), a behavior important for its survival and bloom formation. However, the single-cell mechanisms governing this migration remain unclear. In this study, we investigated the swimming characteristics of individual C. marina cells during day (light) and night (dark) phases. We observed a strong positive correlation between the length of the propulsive anterior flagellum and the cells swimming speed. We discovered that the length distribution of the anterior flagellum is different during the day and at night. We also found that the beat frequency of the anterior flagellum was significantly higher during the day compared to the night. This resulted in faster mean swimming speeds during the light phase. To investigate the mechanism of length regulation, we tested the role of intraflagellar transport (IFT) using the IFT dynein inhibitor, ciliobrevin D. Treatment with ciliobrevin D induced a time- and concentration-dependent shortening of the anterior flagellum. This is the first pharmacological evidence to suggest that an IFT-like mechanism may actively control motile flagellar length in C. marina. These findings suggest that C. marina modulates its swimming speed through diurnal changes in both flagellar length and beat frequency, likely as an energy-saving strategy coupled to its DVM.

1Diatoms are microscopic marine algae that are critical for global primary production, carbon sequestration, and fisheries productivity. However, select diatoms may form harmful algal blooms, which threaten marine ecosystems and the fisheries they sustain. Rapidly identifying harmful blooms is necessary to effectively manage marine resources, yet current identification methods are limited by expensive and labor-intensive in situ point sampling. Hyperspectral remote sensing enables scalable monitoring, but its ability to resolve taxonomic shifts within phytoplankton groups (e.g. diatoms) is largely unknown. To investigate this uncertainty, we cultured four dominant diatom genera from the California Current upwelling system, including this systems most abundant harmful algae, Pseudo-nitzschia. The hyperspectral absorption and backscatter of these taxa were measured and used to model spectral reflectances that remote sensing platforms (satellites/drones) might detect. Differences between fingerprints of these taxa were quantified using vector-based and statistical analyses. Mean spectral differences of 48% were observed between the most dominant diatom, Thalassiosira, and the most toxic diatom, Pseudo-nitzschia. Differences of approximately 30% were found between Pseudo-nitzschia and the second and third most abundant diatoms, Chaetoceros and Asterionellopsis. Successful identification of Pseudo-nitzschias reflectance fingerprint was driven by the presence of a unique feature around 560 nm. The distinct spectral fingerprint of Pseudo-nitzschia indicates that it can be distinguished from benign diatom blooms using hyperspectral remote sensing platforms.

The transcription factor Pho4 is crucial for the response to phosphate starvation in many fungi, and it has been linked to tolerance to alkalinization of the medium and to pathogenicity. It is widely accepted that it is encoded by a single gene. However, the industrially relevant yeast Komagataella phaffii might contain two Pho4-encoding genes (PAS_chr1-1_0265 and PAS_chr2-1_0177, designated here PHO4(A) and PHO4(B), respectively), which have never been functionally characterized. The phenotypic analysis of single and double mutants suggests that Pho4(B) plays a major role in the adaptation to Pi scarcity. While single mutants exhibited limited and non-overlapping phenotypic defects, the pho4(A) pho4(B) strain was sensitive to multiple types of stress, including phosphate starvation and alkaline pH. Transcriptomic analysis confirms that Pho4(B) is crucial for the transcriptional response to phosphate starvation, including induction of typical gene markers (PHO5, PHO89, VTC1, etc.). However, by using a GFP reporter we found that PHO4(A) also participates in the induction of PHO89 under high pH stress. Expression of both PHO4(A) and PHO4(B) in S. cerevisiae complemented the pho4 mutation under phosphate limitation by restoring growth, expression of the Pho84 transporter and secreted phosphatase activity. These results indicate that both transcription factors display partially overlapping functions, responding differently to diverse stimuli, and that together they constitute a key component in the adaptation to a variety of stresses. Therefore, K. phaffii is an exceptional example among fungi that encodes two Pho4 functional transcription factors.