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Effects of biostimulants, AMPEP and Kelpak on the growth and asexual reproduction of Pyropia yezoensis (Bangiales, Rhodophyta) at different temperatures

Acadian marine plant extract powder (AMPEP) and Kelpak are commercial biostimulants derived from brown algae Ascophyllum nodosum. This study was to determine if AMPEP and Kelpak can induce thermal resistance in Pyropia yezoensis. P. yezoensis blades were exposed to different concentrations (control: 0, low: 0.001, high: 1 ppm) of AMPEP and Kelpak at 10°C for 6 and 7 days, respectively. Those blades were then cultivated in von Stosch enriched seawater medium at different temperatures (10, 15, 20, and 25°C) with 12 : 12 L : D photoperiod and 100 μmol m-2 s-1 of photosynthetically active radiation for additional 15 days. Results showed that P. yezoensis reproduced archeospores at 20 and 25°C at all biostimulant conditions within 15 days. At lower temperatures (10 and 15°C), only AMPEP-treated P. yezoensis reproduced archeospores. P. yezoensis exposed to 1 ppm Kelpak exhibited higher phycoerythrin and phycocyanin contents than control and 0.001 ppm conditions at 15°C. AMPEP-treated conditions showed higher phycoerythrin and phycocyanin contents than control at 10°C. These results suggest that AMPEP and Kelpak may not enhance the thermal resistance of P. yezoensis. However, AMPEP stimulated archeospores release at lower temperatures. The treatment of AMPEP and Kelpak also increased the pigment contents in P. yezoensis. These results suggest that the use of seaweed-derived biostimulants can provide some economic benefits in P. yezoensis aquaculture. The enhancement of archeospores formation by AMPEP at lower temperature may also increase the productivity since Pyropia farming relies on the accumulation of secondary seedings via asexual reproduction.

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Physiological and transcriptome analysis of acclimatory response to cold stress in marine red alga Pyropia yezoensis

Red macroalga Pyropia yezoensis is a high valuable cultivated marine crop. Its acclimation to cold stress is especially important for long cultivation period across winter in coasts of warm temperate zone in East Asia. In this study, the response of P. yezoensis thalli to low temperature was analyzed on physiology and transcriptome level, to explore its acclimation mechanism to cold stress. The results showed that the practical photosynthesis activity (indicated by ΦPSII and qP) was depressed and pigment allophycocyanin content was decreased during the cold stress of 48 h. However, the Fv/Fm and non-photochemical quenching increased significantly after 24 h, and the average growth rate of thalli also rebounded from 24 to 48 h, indicating a certain extent of acclimation to cold stress. On transcriptionally, the low temperature promoted the expression of differentially expressed genes (DEGs) related to carbohydrate metabolism and energy metabolism, while genes related to photosynthetic system were depressed. The increased expression of DEGs involved in ribosomal biogenesis and lipid metabolism which could accelerate protein synthesis and enhance the degree of fatty acid unsaturation, might help P. yezoensis thallus cells to cope with cold stress. Further co-expression network analysis revealed differential expression trends along with stress time, and corresponding hub genes play important roles in the systemic acquired acclimation to cold stress. This study provides basic mechanisms of P. yezoensis acclimation to cold temperature and may aid in exploration of functional genes for genetic breeding of economic macroalgae.

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Holocarpic oomycete parasites of red algae are not Olpidiopsis, but neither are they all Pontisma or Sirolpidium (Oomycota)

Oomycetes are ubiquitous heterotrophs of considerable economic and ecological importance. Lately their diversity in marine environments has been shown to be greatly underappreciated and many lineages of intracellular holocarpic parasites, infecting micro- and macro-algae, remain to be fully described taxonomically. Among them, pathogens of marine red algae have been studied extensively as they infect important seaweed crops. Throughout the 20th century, most intracellular, holocarpic biotrophic oomycetes that infect red algae have been assigned to the genus Olpidiopsis Cornu. However, 18S rRNA sequencing of Olpidiopsis saprolegniae, the species considered the generitype for Olpidiopsis, suggests that this genus is not closely related to the marine pathogens and that the latter requires a nomenclatural update. Here, we compile and reanalyze all recently published 18S rRNA sequence data for marine holocarpic oomycetes, with a particular focus on holocarpic pathogens of red algae. Their taxonomy has been revised twice over the past four years, with suggestions to transfer them first into the genus Pontisma and then Sirolpidium, and into a monogeneric order, Pontismatales. We show however, that previously published topologies and the proposed taxa Pontisma, Sirolpidium, and Pontismatales are unsupported. We highlight that name changes that are unfounded and premature create confusion in interested parties, especially concerning pathogens of marine red algae that infect important seaweed crops. We thus propose that the names of these holocarpic biotrophic parasites of red algae are retained temporarily, until a supported topology is produced with more genetic markers to enable the circumscription of species and higher-level taxa.

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Estimation of bioluminescence intensity of the dinoflagellates Noctiluca scintillans, Polykrikos kofoidii, and Alexandrium mediterraneum populations in Korean waters using cell abundance and water temperature

Many dinoflagellates produce bioluminescence. To estimate the intensity of bioluminescence produced by populations of the heterotrophic dinoflagellates Noctiluca scintillans and Polykrikos kofoidii and autotrophic dinoflagellate Alexandrium mediterraneum in Korean waters, we measured cellular bioluminescence intensity as a function of water temperature and calculated population bioluminescence intensity with cell abundances and water temperature. The mean 200-second-integrated bioluminescence intensity per cell (BLcell) of N. scintillans satiated with the chlorophyte Dunaliella salina decreased continuously with increasing water temperature from 5 to 25°C. However, the BLcell of P. kofoidii satiated with the mixotrophic dinoflagellate Alexandrium minutum continuously increased from 5 to 15°C but decreased at temperatures exceeding this (to 30°C). Similarly, the BLcell of A. mediterraneum continuously increased from 10 to 20°C but decreased between 20 and 30°C. The difference between highest and lowest BLcell of N. scintillans, P. kofoidii, and A. mediterraneum at the tested water temperatures was 3.5, 11.8, and 21.0 times, respectively, indicating that water temperature clearly affected BLcell. The highest estimated population bioluminescence intensity (BLpopul) of N. scintillans in Korean waters in 1998–2022 was 4.22 × 1013 relative light unit per liter (RLU L-1), which was 1,850 and 554,000 times greater than that of P. kofoidii and A. mediterraneum, respectively. This indicates that N. scintillans populations produced much brighter bioluminescence in Korean waters than the populations of P. kofoidii or A. mediterraneum.

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Oomycete pathogens, red algal defense mechanisms and control measures

Oomycete pathogens are one of the most serious threats to the rapidly growing global algae aquaculture industry but research into how they spread and how algae respond to infection is unresolved, let alone a proper classification of the pathogens. Even the taxonomy of the genera Pythium and Olpidiopsis, which contain the most economically damaging pathogens in red algal aquaculture, and are among the best studied, needs urgent clarification, as existing morphological classifications and molecular evidence are often inconsistent. Recent studies have reported a number of genes involved in defense responses against oomycete pathogens in red algae, including pattern-triggered immunity and effectortriggered immunity. Accumulating evidence also suggests that calcium-mediated reactive oxygen species signaling plays an important role in the response of red algae to oomycete pathogens. Current management strategies to control oomycete pathogens in aquaculture are based on the high resistance of red algae to abiotic stress, these have environmental consequences and are not fully effective. Here, we compile a revised list of oomycete pathogens known to infect marine red algae and outline the current taxonomic situation. We also review recent research on the molecular and cellular responses of red algae to oomycete infection that has only recently begun, and outline the methods currently used to control disease in the field.

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Protists in hypoxic waters of Jinhae Bay and Masan Bay, Korea, based on metabarcoding analyses: emphasizing surviving dinoflagellates

Hypoxia can indeed impact the survival of protists, which play a crucial role in marine ecosystems. To better understand the protistan community structure and species that can thrive in hypoxic waters, we collected samples from both the surface and bottom waters during the hypoxic period in Jinhae and Masan Bays and the non-hypoxic period in Jinhae Bay. Subsequently, we utilized metabarcoding techniques to identify the protistan species. During hypoxia, with dissolved oxygen concentrations of 0.8 mg L<sup>-1</sup> in Jinhae Bay and 1.8 mg L<sup>-1</sup> in Masan Bay within the bottom waters, the phylum Dinoflagellata exhibited the highest amplicon sequence variants richness among the identified protist phyla. Following the Dinoflagellata, Ochrophyta and Ciliophora also displayed notable presence. In hypoxic waters of Jinhae and Masan Bays, we identified a total of 36 dinoflagellate species that exhibited various trophic modes. These included one autotrophic species, 14 mixotrophic species, 9 phototrophic species with undetermined trophic modes (either autotrophic or mixotrophic), 2 kleptoplastidic species, and 10 heterotrophic species. Furthermore, the hypoxic bottom water exhibited a greater number of heterotrophic dinoflagellate species compared to the non-hypoxic surface water within the same water column or the non-hypoxic bottom water. Therefore, feeding by mixotrophic and heterotrophic dinoflagellates may be partially responsible for their dominance in terms of the number of species surviving in hypoxic waters. This study not only introduces the initial documentation of 26 dinoflagellate species surviving in hypoxic conditions but also establishes a foundation for a more comprehensive understanding of the ecophysiology of dinoflagellates in hypoxic marine environments.

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Antiviral effect of fucoxanthin obtained from Sargassum siliquastrum (Fucales, Phaeophyceae) against severe acute respiratory syndrome coronavirus 2

Human coronavirus diseases, particularly severe acute respiratory syndrome coronavirus 2, still remain a persistent public health issue, and many recent studies are focusing on the quest for new leads against coronaviruses. To contribute to this growing pool of knowledge and explore the available marine natural products against coronaviruses, this study investigated the antiviral effects of fucoxanthin isolated from Sargassum siliquastrum—a brown alga found on Jeju Island, South Korea. The antiviral effects of fucoxanthin were confirmed in severe acute respiratory syndrome coronavirus 2-infected Vero cells, and its structural characteristics were verified in silico using molecular docking and molecular dynamic simulations and in vitro colorimetric method. Fucoxanthin inhibited the infection in a concentration-dependent manner, without showing cytotoxicity. Molecular docking simulations revealed that fucoxanthin binds to the angiotensinconverting enzyme 2-spike protein (binding energy -318.306 kcal mol<sup>-1</sup>) and main protease (binding energy -205.118 kcal mol<sup>-1</sup>). Moreover, molecular dynamic simulations showed that fucoxanthin remains docked to angiotensin-converting enzyme 2-spike protein for 20 ns, whereas it breaks away from main protease after 3 ns. Also, the in silico prediction of the fucoxanthin was verified through the in vitro colorimetric method by inhibiting the binding between angiotensinconverting enzyme 2 and spike protein in a concentration-dependent manner. These results indicate that fucoxanthin exhibits antiviral effects against severe acute respiratory syndrome coronavirus 2 by blocking the entry of the virus. Therefore, fucoxanthin from S. siliquastrum can be a potential candidate for treating coronavirus infection.

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Morphology and phylogenetic relationships of two Antarctic strains within the genera Carolibrandtia and Chlorella (Chlorellaceae, Trebouxiophyceae)

The genera Carolibrandtia and Chlorella have been described as small green algae with spherical cell shapes that inhabit various environments. Species of these genera are often difficult to identify because of their simple morphology and high phenotypic plasticity. We investigated two small coccoid strains from Antarctica based on morphology, molecular phylogeny by two alignment methods which have been applied to previous phylogenetic studies of the genus Chlorella, and comparison of the secondary structures of nuclear small subunit (SSU) and internal transcribed spacer (ITS) rDNA sequences. Light microscopy of two strains revealed spherical cells containing chloroplasts with pyrenoids, and the morphological characteristics of the strains were nearly identical to those of other Chlorella species. However, based on the phylogenetic analyses of nuclear SSU and ITS rDNA sequences, it was determined that the Antarctic microalgal strains belonged to two genera, as the Chlorella and Carolibrandtia. In addition, the secondary structures of the SSU and ITS2 sequences were analyzed to detect compensatory base changes (CBCs) that were used to identify and describe the two strains. A unique CBC in the SSU rDNA gene was decisive for distinguishing strain CCAP 211/45. The ITS2 rDNA sequences for each strain were compared to those obtained previously from other closely related species. Following the comparison of morphological and molecular characteristics, we propose KSF0092 as a new species, Chlorella terrestris sp. nov., and the reassignment of the strain Chlorella antarctica CCAP 211/45 into Carolibrandtia antarctica comb. nov.

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