Diatoms are unicellular algae that are characterized by their silica cell walls. The silica elements form intracellularly in a membrane-bound organelle, and are exocytosed after completion. How diatoms maintain membrane homeostasis during the exocytosis of these large and rigid silica elements is a long-standing enigma. We studied membrane dynamics during cell wall formation and exocytosis in the diatom Stephanopyxis turris, using live-cell confocal microscopy and advanced electron microscopy. Our results provide detailed information on the ultrastructure and dynamics of the silicification process, showing that during cell wall formation, the organelle membranes tightly enclose the mineral phase, creating a precise mold of the delicate geometrical patterns. Surprisingly, during exocytosis of the mature silica elements, the proximal organelle membrane becomes the new plasma membrane, and the distal membranes gradually disintegrate into the extracellular space without any noticeable endocytic retrieval or extracellular repurposing. These observations suggest that diatoms evolved an extraordinary exocytosis mechanism in order to secrete their cell wall elements.

Coccolithophores are a group of unicellular marine algae that shape global geochemical cycles via the production of calcium carbonate crystals. Interestingly, different life‐cycle phases of the same coccolithophore species produce very different calcitic scales, called coccoliths. In the widely studied diploid phase, the crystals have anisotropic and complex morphologies, while haploid cells produce coccoliths consisting solely of calcite crystals with simple rhombohedral morphology. Understanding how these two life‐cycle phases control crystallization is a highly sought‐after goal, yet, haploid phase crystallization has rarely been studied, and the process by which they form is unknown. Herein, advanced electron microscopy is employed to elucidate the cellular architecture of the calcification process in haploid cells. The results show that in contrast to diploid‐phase calcification, the coccolith‐forming vesicle of haploid‐phase cells is voluminous. In this solution‐like environment, the crystals nucleate and grow asynchronously in a process that resembles calcite growth in bulk solution, leading to the simple morphologies of the crystals. The two distinct mineralization regimes of coccolithophore life‐cycle phases suggest that cellular architecture, and specifically confinement of the crystallization process, is a pivotal determinant of biomineral morphology and assembly.

Diatoms are unicellular algae characterized by silica cell walls. These silica elements are known to be formed intracellularly in membrane-bound silica deposition vesicles and exocytosed after completion. How diatoms maintain membrane homeostasis during the exocytosis of these large and rigid silica elements remains unknown. Here we study the membrane dynamics during cell wall formation and exocytosis in two model diatom species, using live-cell confocal microscopy, transmission electron microscopy and cryo-electron tomography. Our results show that during its formation, the mineral phase is in tight association with the silica deposition vesicle membranes, which form a precise mold of the delicate geometrical patterns. We find that during exocytosis, the distal silica deposition vesicle membrane and the plasma membrane gradually detach from the mineral and disintegrate in the extracellular space, without any noticeable endocytic retrieval or extracellular repurposing. We demonstrate that within the cell, the proximal silica deposition vesicle membrane becomes the new barrier between the cell and its environment, and assumes the role of a new plasma membrane. These results provide direct structural observations of diatom silica exocytosis, and point to an extraordinary mechanism in which membrane homeostasis is maintained by discarding, rather than recycling, significant membrane patches.

Unicellular organisms are known to exert tight control over their cell size. In the case of diatoms, abundant eukaryotic microalgae, the layout of the rigid silica cell wall imposes geometrical restrictions on cell size. A generally accepted theory states that the need to fit any new silica element into a previously formed structure causes a reduction in size with each vegetative division cycle, until cell size restoration is achieved by a switch to another life-cycle stage. Nevertheless, several reported exceptions cast doubt on the generality of this theory. Here, we monitored clonal cultures of the diatom Stephanopyxis turris for up to two years, recording the sizes of thousands of cells, in order to follow the distribution of cell sizes in the population. Our results show that all S. turris cultures above a certain size threshold undergo a gradual size reduction, in accordance with the postulated geometrical driving force. However, once the cell size reaches a lower threshold, a constant size range is maintained by different cellular strategies. These observations suggest two distinct mechanisms to regulate the cell size of diatoms, reduction and homeostasis. The interplay between these mechanisms can explain the behavior of different diatoms species in various environments.