Yinjie Zhu

Harmful algal blooms, the most severe ecological hazards worldwide, terminate abruptly within a few days. In this work, we identified that iron-catalyzed active lipid peroxides predominantly trigger individual cell ferroptosis and drive the population collapse of blooming cyanobacteria. We reveal the chronological sequence of labile iron burst, oxidative stress, lipid peroxidation, and cell death during a Microcystis bloom demise event. Dead cells exhibit a nonrandom spatial distribution within colonies. Intensifying lipid peroxidation catalyzed by cellular labile iron generates truncated phospholipids with shortened fatty acyl chains bearing alkyl groups. These active lipid peroxides destabilize plasma membranes and induce nanoscale membrane pore formation, resulting in individual cell ferroptosis and lysis. Oxidized lipids are also released from ferroptotic cells, propagating lipid peroxidation to neighboring cells, thereby spreading death throughout the population.

Single-cell sequencing (SCS) is an evolutionary technique for conducting life science research, providing the highest genome-sale throughput and single-cell resolution and unprecedented capabilities in addressing mechanistic and operational questions. Unfortunately, the current SCS pipeline cannot be directly applied to algal research as algal cells have cell walls, which makes RNA extraction hard for the current SCS platforms. Fortunately, effective methods are available for producing algal protoplasts (cells without cell walls), which can be directly fed into current SCS pipelines. In this review, we first summarize the cell wall structure and chemical composition of algal cell walls, particularly in Chlorophyta, then summarize the advances made in preparing algal protoplasts using physical, chemical, and biological methods, followed by specific cases of algal protoplast production in some commonly used eukaryotic algae. This review provides a timely primer to those interested in applying SCS in eukaryotic algal research.

Bloom-forming cyanobacteria produce and release odorous compounds and pose threats to the biodiversity of aquatic ecosystem and to the drinking water supply. In this study, the concentrations of β-cyclocitral in different bacterial growth phases were investigated using GC–MS to determine the growth stage of Microcystis aeruginosa at high risk for β-cyclocitral production. Moreover, the synchronicity of the production of β-cyclocitral and its precursor β-carotene at both population and single-cell levels was assessed. The results indicated that β-cyclocitral was the main odorous compound produced by M. aeruginosa cells. The intracellular concentration of β-cyclocitral (C_β-cc) as well as its cellular quota (Q_β-cc) increased synchronously in the log phase, along with the increase of cell density. However, they reached the maximum values of 415 μg/L and 10.7 fg/cell in the late stationary phase and early stationary phase, respectively. The early stage of the stationary phase is more important for β-cyclocitral monitoring, and the sharp increase in Q_β-cc is valuable for anticipating the subsequent increase in C_β-cc. The molar concentrations of β-cyclocitral and β-carotene showed a linear relationship, with an R² value of 0.92, suggesting that the production of β-cyclocitral was linearly dependent on that of β-carotene, especially during the log phase. However, the increase in Q_β-cc was slower than that in β-carotene during the stationary phase, suggesting that β-cyclocitral production turned to be carotene oxygenase-limited when the growth rate decreased. These results demonstrate that variations of β-cyclocitral production on a single-cell level during different bacterial growth phases should be given serious consideration when monitoring and controlling the production of odorous compounds by M. aeruginosa blooms.