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Diatoms are single-celled photosynthetic eukaryotes responsible for CO2 fixation and primary production in aquatic ecosystems. The cosmopolitan marine diatom Coscinodiscus granii can form seasonal blooms in coastal areas and interact with various microorganisms, including the parasitic oomycete Lagenisma coscinodisci. This unicellular eukaryote is mainly present in the northern hemisphere as an obligate parasite of the genus Coscinodiscus. Understanding the interplay of abiotic factors such as temperature and biotic factors like parasitism on algal physiology is crucial as it dictates plankton community composition and is especially relevant during environmental changes and warming events. This study investigates the impact of two temperatures, 13 °C and 25 °C, on Coscinodiscus granii under laboratory conditions. A decreased infection rate of the parasite was observed at the elevated temperature. Comparative metabolomic analysis using UHPLC-HRMS revealed that temperature and parasitism significantly affect the algal cell metabolome. Abundances of metabolites related to sulfur metabolism, including cysteinoleic acid and dimethylsulfoniopropionate, as well as molecules linked to fatty acid metabolism, e.g., carnitine, acetylcarnitine, and eicosapentanoic acid, significantly increase in cells grown at a higher temperature, suggesting the enhanced rate of metabolism of host cells as the temperature rises. Our study reveals how temperature-induced metabolic changes can influence host–parasite dynamics in a changing environment. [ABSTRACT FROM AUTHOR]

Plankton parasites such as viruses, bacteria, fungi, and unicellular eukaryotes are associated symbionts colonizing algal groups in aquatic ecosystems. They occur within a network of microbe–microbe interactions in which they parasitize phytoplankton and seaweeds, i.e., primary producers generating organic carbon and forming the basis of marine food webs. These parasites use algae as a source of nutrients and reproduce at the expense of their host, causing infectious symptoms leading to disease and death. Plankton parasites can reduce the algal populations, and the infection of one specific species may, in turn, favor the development of another, influencing the seasonal succession of algal blooms in oceans, seas, and lakes. Many parasites have free-living stages that zooplankton graze upon, representing a significant trophic link in food webs. The biology and life cycle of plankton parasites are well investigated in marine and freshwater algal hosts. Still, the chemical signaling mediating these microbial interactions and the effect of co-occurring symbionts remain underexplored. This review focuses on the diversity of plankton parasites infecting algae, with a particular emphasis on unicellular eukaryotes. The ecological role of plankton parasites, the mechanisms regulating cellular infection and host resistance, and the interplay of alga-parasite interactions with coexisting microorganisms are discussed. [ABSTRACT FROM AUTHOR]

Bloom-forming phytoplankton are key players in aquatic ecosystems, fixing carbon dioxide and forming the base of the marine food web. Diverse stresses, such as nutrient depletion, temperature increase, and pathogen emergence can influence the health and dynamics of algal populations. While population responses to these stressors are welldocumented in the aquatic ecosystems, little is known about the individual cellular adaptations. These are however the key to an in-depth physiological understanding of microbiome dynamics in the plankton. Finding solutions to disease control in aquaculture also depends on knowledge of infection dynamics and physiology in algae. Single-cell metabolomics can give insight into infection processes by providing a snapshot of small molecules within a biological system. We used a single-cell metabolome profiling workflow to track metabolic changes of diatoms and dinoflagellates subjected to parasite infection caused by the oomycete Lagenisma coscinodisci and the alveolate Parvilucifera spp. We accurately classified the healthy phenotype of bloom-forming phytoplankton, including the diatoms Coscinodiscus granii and Coscinodiscus radiatus, and the toxic dinoflagellate Alexandrium minutum. We discriminated the infection of the toxic dinoflagellate A. minutum with the alveolate parasitoids Parvilucifera infectans and P. rostrata down to the single-cell resolution. Strain and species-specific responses of the diatom hosts Coscinodiscus spp. Infected with the oomycete pathogen Lagenisma coscinodisci could be recognized. LC-HRMS and fragmentation pattern analysis enabled the structure elucidation of metabolic predictors of infection (guanine, xanthine, DMSP, and pheophorbide). The purine salvage pathway and DMSP lysis could be assigned as regulated processes during host invasion. The findings establish single-cell metabolome profiling with LDI-HRMS coupled with classification analysis as a reliable diagnostic tool to track metabolic changes in algae. [ABSTRACT FROM AUTHOR]

Introduction: Marine planktonic communities are complex microbial consortia often dominated by microscopic algae. The taxonomic identification of individual phytoplankton cells usually relies on their morphology and demands expert knowledge. Recently, a live single-cell mass spectrometry (LSC-MS) pipeline was developed to generate metabolic profiles of microalgae. Objective: Taxonomic identification of diverse microalgal single cells from collection strains and plankton samples based on the metabolic fingerprints analyzed with matrix-free laser desorption/ionization high-resolution mass spectrometry. Methods: Matrix-free atmospheric pressure laser-desorption ionization mass spectrometry was performed to acquire single-cell mass spectra from collection strains and prior identified environmental isolates. The computational identification of microalgal species was performed by spectral pattern matching (SPM). Three similarity scores and a bootstrap-derived confidence score were evaluated in terms of their classification performance. The effects of high and low-mass resolutions on the classification success were evaluated. Results: Several hundred single-cell mass spectra from nine genera and nine species of marine microalgae were obtained. SPM enabled the identification of single cells at the genus and species level with high accuracies. The receiver operating characteristic (ROC) curves indicated a good performance of the similarity measures but were outperformed by the bootstrap-derived confidence scores. Conclusion: This is the first study to solve taxonomic identification of microalgae based on the metabolic fingerprints of the individual cell using an SPM approach. [ABSTRACT FROM AUTHOR]

Brown macroalgae are an essential component of temperate coastal ecosystems and a growing economic sector. They harbor diverse microbial communities that regulate algal development and health. This algal holobiont is dynamic and achieves equilibrium via a complex network of microbial and host interactions. We now report that bacterial and fungal endophytes associated with four brown algae (Ascophyllum nodosum , Pelvetia canaliculata , Laminaria digitata , and Saccharina latissima) produce metabolites that interfere with bacterial autoinducer-2 quorum sensing, a signaling system implicated in virulence and host colonization. Additionally, we performed co-culture experiments combined to a metabolomic approach and demonstrated that microbial interactions influence production of metabolites, including metabolites involved in quorum sensing. Collectively, the data highlight autoinducer-2 quorum sensing as a key metabolite in the complex network of interactions within the algal holobiont. [ABSTRACT FROM AUTHOR]

Unicellular phototrophic algae can form massive blooms with up to millions of individual cells per milliliter in freshwater and marine ecosystems. Despite the temporal dominance of bloom formers many algal species can co-exist and compete for nutrients and space, creating a complex and diverse community. While microscopy and single cell genomics can address the taxonomic inventory, the cellular metabolome has yet to be thoroughly explored to determine the physiological status of microalgae. This might, however, provide a key to understand the observed species diversity in the homogeneous environment. Here, we introduce an effective, rapid and versatile method to analyze living single cells from aqueous substrata with laser-desorption/ionization mass spectrometry (LDI-MS) using a simple and inexpensive matrix-free support. The cells deposited on a cultivation-medium wetted support are analyzed with minimal disturbance as they remain in their natural viable state until their disruption during LDI-MS. Metabolites desorbed from single cells are analyzed on High-Resolution Mass Spectrometry (HR-MS) using the Orbitrap FT-MS technology to fingerprint cellular chemistry. This live single-cell mass spectrometry (LSC-MS) allows assessing the physiological status and strain-specifics of different microalgae, including marine diatoms and freshwater chlorophytes, at the single-cell level. We further report a reliable and robust data treatment pipeline to perform multivariate statistics on the replicated LSC-MS data. Comparing single cell MS spectra from natural phytoplankton samples and from laboratory strains allows the identification and discrimination of inter and intra-specific metabolic variability and thereby has promising applications in addressing highly complex phytoplankton communities. Notably, the herein described matrix-free live-single-cell LDI-HR-MS approach enables monitoring dynamics of the plankton and might explain why key-players survive, thrive, avoid selective feeding or pathogenic virus and bacteria, while others are overcome and die. [ABSTRACT FROM AUTHOR]

Kelps are colonized by a wide range of microbial symbionts. Among them, endophytic fungi remain poorly studied, but recent studies evidenced yet their high diversity and their central role in algal defense against various pathogens. Thus, studying the metabolic expressions of kelp endophytes under different conditions is important to have a better understanding of their impacts on host performance. In this context, fatty acid composition is essential to a given algae fitness and of interest to food web studies either to measure its nutritional quality or to infer about its contribution to consumers diets. In the present study, Paradendryphiella salina, a fungal endophyte was isolated from Saccharina latissima (L.) and Laminaria digitata (Hudson.) and its fatty acid composition was assessed at increasing salinity and temperature conditions. Results showed that fungal composition in terms of fatty acids displayed algal-dependent trajectories in response to temperature increase. This highlights that C18 unsaturated fatty acids are key components in the host-dependant acclimation of P. salina to salinity and temperature changes. [ABSTRACT FROM AUTHOR]

High-throughput screening assays have been designed to identify compounds capable of inhibiting phenotypes involved in cancer aggressiveness. However, most studies used commercially available chemical libraries. This prompted us to explore natural products isolated from marine-derived fungi as a new source of molecules. In this study, we established a chemical library from 99 strains corresponding to 45 molecular operational taxonomic units and evaluated their anticancer activity against the MCF7 epithelial cancer cell line and its invasive stem cell-like MCF7-Sh-WISP2 counterpart. We identified the marine fungal Paradendryphiella salina PC 362H strain, isolated from the brown alga Pelvetia caniculata (PC), as one of the most promising fungi which produce active compounds. Further chemical and biological characterizations of the culture of the Paradendryphiella salina PC 362H strain identified (-)-hyalodendrin as the active secondary metabolite responsible for the cytotoxic activity of the crude extract. The antitumor activity of (-)-hyalodendrin was not only limited to the MCF7 cell lines, but also prominent on cancer cells with invasive phenotypes including colorectal cancer cells resistant to chemotherapy. Further investigations showed that treatment of MCF7-Sh-WISP2 cells with (-)-hyalodendrin induced changes in the phosphorylation status of p53 and altered expression of HSP60, HSP70 and PRAS40 proteins. Altogether, our study reveals that this uninvestigated marine fungal crude extract possesses a strong therapeutic potential against tumor cells with aggressive phenotypes and confirms that members of the epidithiodioxopiperazines are interesting fungal toxins with anticancer activities. [ABSTRACT FROM AUTHOR]

An amendment to this paper has been published and can be accessed via a link at the top of the paper. [ABSTRACT FROM AUTHOR]

Flagellated oomycetes frequently infect unicellular algae, thus limiting their proliferation. Here we show that the marine oomycete Lagenisma coscinodisci rewires the metabolome of the bloom-forming diatom Coscinodiscus granii, thereby promoting infection success. The algal alkaloids β-carboline and 4-carboxy-2,3,4,9-tetrahydro-1H-β-carboline are induced during infection. Single-cell profiling with AP-MALDI-MS and confocal laser scanning microscopy reveals that algal carbolines accumulate in the reproductive form of the parasite. The compounds arrest the algal cell division, increase the infection rate and induce plasmolysis in the host. Our results indicate that the oomycete manipulates the host metabolome to support its own multiplication. Flagellated oomycetes frequently infect unicellular algae, thus limiting their proliferation. Here, the authors show that an oomycete rewires the metabolome of a marine bloom-forming diatom, thereby promoting infection success. [ABSTRACT FROM AUTHOR]