Shinya Yoshikawa

Parmales (Bolidophyceae), the closest relatives of diatoms, includes isolates exhibiting one of two distinct morphotypes: a silicified non-flagellated form (S-type) and a naked flagellated form (F-type). Although alternation of these two forms for a single isolate has not been formally established, previous studies hypothesized that these morphotypes represent alternating stages of a single parmalean. In this study, we investigated the global expression patterns of S- and F-type marker genes by integrating parmalean Metagenome-Assembled Genomes (MAGs) and the metatranscriptomic dataset from Tara Oceans. We detected the expression of both S- and F-type marker genes from individual environmental MAGs. This finding provides the first metatranscriptomic evidence that natural parmalean genomes possess the potential to manifest both morphotypes. Furthermore, our analysis revealed different geographical expression patterns between the two forms. The expression of F-type markers showed a broad distribution, whereas that of S-type markers was more restricted, suggesting distinct niches for the two morpho-phases. Moreover, S-type gene expression appears to require specific environmental triggers that lead to a higher population density, whereas F-type expression is rather constitutively maintained. Overall, our results support the hypothesis of a life cycle involving morphological switching and reconcile the long-standing discrepancy between the ubiquity of parmaleans in molecular surveys and the limited geographical range for the observation of silicified cells. Based on these patterns, we propose a threshold-based model in which F-type-dominated populations persist under conditions unfavorable for growth and a morphological switch to the S-type is triggered once environmental conditions exceed a critical threshold for growth.

SUMMARY: Understanding the environmental factors that influence the release of eggs and swarmers, including gametes, zoospores and unsexual swimming cells, is essential for deciphering the survival strategies of brown algae. In the present study, we investigated the mechanisms underlying the light‐induced release of swarmers in Petalonia fascia. We found that swarmer release was triggered by light, especially blue and green light, with red light having a significantly smaller effect. Swarmer release was not directly affected by photosynthesis. In addition, the duration of light exposure was crucial: even when the total number of photons was low, a longer exposure time resulted in a higher release rate of swarmers. This result suggests that a minimum light exposure time is necessary for swarmer release. We also investigated the influence of a circadian rhythm on swarmer release. Under continuous light conditions, swarmers were released at subjective dawn, thus indicating a circadian rhythm influence. By contrast, no release occurred at subjective dawn in continuous darkness. This result suggests that light is necessary to trigger release, even when circadian rhythms are involved. Our findings, which demonstrate that light quality, exposure time and circadian regulation are critical factors for swarmer release in P. fascia, provide new insights into how environmental cues influence reproductive processes in brown algae.

In this study, we investigated the photoperiodic responses regulating erect thallus formation in Petalonia fascia (KU-1293). We found that, through critical day length analysis and night break treatment culture experiments, P. fascia formed erect thalli under short-day conditions, indicating a genuine photoperiodic response. The critical day length for this morphological change was 10–11 h. Notably, night-break treatment with blue light (BL) and green light (GL) inhibited, whereas that with red light (RL) enhanced erect thallus formation. P. fascia was more sensitive to BL than to GL, requiring approximately 100-times less light to exhibit similar effects. Furthermore, promotion of erect thallus formation by RL was not negated by far-red light, suggesting the presence of a novel RL receptor distinct from the classical phytochrome system in terrestrial plants. These findings highlight the complexity of light wavelength interactions in regulating photoperiodic responses and suggest the presence of unique photoreceptors for day-length perception and erect thallus formation in P. fascia.

Adenosine 3′,5′-cyclic monophosphate (cAMP) is a universal signaling molecule that acts as a second messenger in various organisms. It is well established that cAMP plays essential roles across the tree of life, although the function of cAMP in land plants has long been debated. We previously identified the enzyme with both adenylyl cyclase (AC) and cAMP phosphodiesterase (PDE) activity as the cAMP-synthesis/hydrolysis enzyme COMBINED AC with PDE (CAPE) in the liverwort Marchantia polymorpha . CAPE is conserved in streptophytes that reproduce with motile sperm; however, the precise function of CAPE is not yet known. In this study, we demonstrate that the loss of function of CAPE in M. polymorpha led to male infertility due to impaired sperm flagellar motility. We also found that two genes encoding the regulatory subunits of cAMP-dependent protein kinase (PKA-R) were also involved in sperm motility. Based on these findings, it is evident that CAPE and PKA-Rs act as a cAMP signaling module that regulates sperm motility in M. polymorpha . Therefore, our results have shed light on the function of cAMP signaling and sperm motility regulators in land plants. This study suggests that cAMP signaling plays a common role in plant and animal sperm motility.

Spermatogenesis is one of the most dramatic changes in cell differentiation. Remarkable chromatin condensation of the nucleus is observed in animal, plant, and algal sperm. Sperm nuclear basic proteins (SNBPs), such as protamine and sperm-specific histone, are involved in chromatin condensation of the sperm nucleus. Among brown algae, sperm of the oogamous Fucales algae have a condensed nucleus. However, the existence of sperm-specific SNBPs in Fucales algae was unclear. Here, we identified linker histone (histone H1) proteins in the sperm and analyzed changes in their gene expression pattern during spermatogenesis in Sargassum horneri. A search of transcriptomic data for histone H1 genes in showed six histone H1 genes, which we named ShH1.1a, ShH1b, ShH1.2, ShH1.3, ShH1.4, and ShH1.5. Analysis of SNBPs using SDS-PAGE and LC–MS/MS showed that sperm nuclei contain histone ShH1.2, ShH1.3, and ShH1.4 in addition to core histones. Both ShH1.2 and ShH1.3 genes were expressed in the vegetative thallus and the male and female receptacles (the organs producing antheridium or oogonium). Meanwhile, the ShH1.4 gene was expressed in the male receptacle but not in the vegetative thallus and female receptacles. From these results, ShH1.4 may be a sperm-specific histone H1 of S. horneri.

The order Parmales (Bolidophyceae) is a minor group of pico-sized eukaryotic marine phytoplankton that contains species with cells surrounded by silica plates. Previous studies revealed that Parmales is a member of ochrophytes and sister to diatoms (Bacillariophyta), the most successful phytoplankton group in the modern ocean. Therefore, parmalean genomes can serve as a reference to elucidate both the evolutionary events that differentiated these two lineages and the genomic basis for the ecological success of diatoms vs. the more cryptic lifestyle of parmaleans. Here, we compared the genomes of eight parmaleans and five diatoms to explore their physiological and evolutionary differences. Parmaleans were predicted to be phago-mixotrophs. By contrast, diatoms have undergone loss of genes related to phagocytosis, indicating the ecological specialization from phago-mixotroph to photoautotroph in the early evolution of diatoms. Furthermore, diatoms showed significant enrichment in gene sets involved in silica metabolism, nutrient uptake capacity, carbon concentrating mechanisms, and iron uptake in comparison with parmaleans. Overall, our results suggest a strong evolutionary link between the loss of phago-mixotrophy and specialization to a silicified photoautotrophic life stage early in diatom evolution after diverging from the Parmales lineage.

SUMMARY: To understand the high temperature stress acclimation of a canopy‐forming kelp, RNA‐seq analysis was performed on cultured Eisenia bicyclis (Kjellman) Setchell (Laminariales, Phaeophyceae), a major component species of kelp forests along the coast of Japan. We established a culture strain from a population at the northernmost distribution limit of this species and measured photosystem II activity in young sporophytes. We found that photosystem II activity was lower at 25°C than at 5–15°C, thus, comparative RNA‐seq analysis was conducted between cultivated young thalli of E. bicyclis at 15°C and 25°C. Comparison with the expression genes revealed 277 up‐regulated genes and 327 down‐regulated genes at 25°C. The most up‐regulated gene was 3‐ketoacyl‐CoA thiolase, which is related to fatty acid degradation. This enzyme has been reported to positively regulate abscisic acid (ABA) signaling in Arabidopsis thaliana (Brassicaceae). In addition, heat shock proteins, ATP synthase, NADH‐dehydrogenase, and L‐ascorbate peroxidase were up‐regulated. The most down‐regulated gene was taurine catabolism dioxygenase TauD/TfdA. Genes related to fatty acid metabolism, photosynthesis, and synthesis of cell wall components were down‐regulated. These results suggest that E. bicyclis may counteract high temperature stress by up‐regulating genes involved in protein stabilization, energy production and antioxidant processes. Conversely, photosynthesis and other metabolic processes may be damaged due to the inability to withstand high temperature stress.

Triparma laevis f. inornata is a unicellular alga belonging to the Bolidophyceae, which is most closely related to diatoms. Like diatoms, T. laevis f. inornata has a siliceous cell wall. The cell wall of T. laevis f. inornata consists of four round plates (three shields and one ventral plate) and one dorsal and three girdle plates. But, unlike diatoms, T. laevis f. inornata cells can grow when concentrations of silica are depleted. We took advantage of this ability, using TEM to study the ontogeny of the siliceous plate, pattern center formation, and development. Two types of pattern centers (annulus and sternum) were observed in the early and middle stage of plate formation. During their formation, the annuli were initially crescent‐shaped but eventually their ends fused to make a ring. Only outward silica deposition of the branching ribs occurred on the growing annulus until it became a ring, resulting in an unfilled circle inside the annulus. The pattern center of the shield plate was always an annulus, but in ventral plates both annulus and sternum were observed. The annuli and sterna in T. laevis f. inornata round plates were very similar to the annuli and sterna in diatom valves. These results suggested that the round plates of Parmales are homologous to diatom valves. This information on the plate ontogeny of T. laevis f. inornata provides new insights into the evolution of the siliceous cell wall in the Parmales and diatoms.

The parmalean algae possess a siliceous wall and represent the sister lineage of diatoms; they are thought to be a key group for understanding the evolution of diatoms. Diatoms possess well-characterized and unique mitotic structures, but the mitotic apparatus of Parmales is still unknown. We observed the microtubule (MT) array during interphase and mitosis in Triparma laevis using TEM. The interphase cells had four or five centrioles (∼80 nm in length), from which MTs emanated toward the cytoplasm. In prophase, the bundle of MTs arose at an extranuclear site. The position of centrioles with respect to an MT bundle changed during its elongation. Centrioles were observed on the lateral side of a shorter MT bundle (∼590 nm) and on either side of an extended MT bundle (∼700 nm). In metaphase, the spindle consisted of two types of MTs-MT bundle that passed through a cytoplasmic tunnel in the center of the nucleus and single MTs (possibly kinetochore MTs) that extended from the poles into the nucleus. The nuclear envelope disappeared only at the regions where the kinetochore MTs penetrated. In telophase, daughter chromosomes migrated toward opposite poles, and the MT bundle was observed between segregating chromosomes. These observations showed that MT nucleation does not always occur at the periphery of centrioles through cell cycle and that the spindle of T. laevis has a similar configuration to that of diatoms.