Light-dependent single-cell heterogeneity in the chloroplast redox state regulates cell fate in a marine diatom

Abstract

Diatoms are photosynthetic microorganisms of great ecological and biogeochemical importance, forming vast blooms in aquatic ecosystems. However, we are still lacking fundamental understanding of how individual cells sense and respond to diverse stress conditions, and what acclimation strategies are employed during bloom dynamics. We investigated cellular responses to environmental stress at the single-cell level using the redox sensor roGFP targeted to various organelles in the diatom Phaeodactylum tricornutum. We detected cell-to-cell variability using flow cytometry cell sorting and a microfluidics system for live imaging of oxidation dynamics. Chloroplast-targeted roGFP exhibited a light-dependent, bi-stable oxidation pattern in response to H2O2 and high light, revealing distinct subpopulations of sensitive oxidized cells and resilient reduced cells. Early oxidation in the chloroplast preceded commitment to cell death, and can be used for sensing stress cues and regulating cell fate. We propose that light-dependent metabolic heterogeneity regulates diatoms’ sensitivity to environmental stressors in the ocean.Microscopic algae, such as diatoms, are widely spread throughout the oceans, and are responsible for half of the oxygen we breathe. At certain times of the year these algae grow very rapidly to form large “blooms” that can be detected by satellites in space. These blooms are generally short-lived because the algae are either eaten by other marine organisms, run out of nutrients, or die as a result of being infected by viruses or bacteria. However, some diatom cells survive the end of the bloom and go on to generate new blooms in the future, but it is still not clear how.As the bloom collapses, diatoms experience many stressful conditions which can cause active molecules known as reactive oxygen species, or ROS for short, to accumulate inside cells. Normally growing cells also produce low amounts of ROS, which regulate various processes that are important for maintaining a cell’s health. However, high amounts of ROS can cause damage, which may lead to a cell’s death. Now, Mizrachi et al. investigated why some algae survive while others die in response to stressful conditions, focusing on the amount of ROS that accumulates within the diatom Phaeodactylum tricornutum.Laboratory experiments showed that individual cells of P. tricornutum respond differently to environmental stress, forming two distinct groups of either sensitive or resilient cells. Sensitive cells accumulated high levels of ROS within a cell compartment known as the chloroplast and eventually died. Whereas resilient cells were able to maintain low levels of ROS in the chloroplast and survived long after the other cells perished. Populations of genetically identical diatom cells also formed distinct groups of sensitive and resilient cells, demonstrating that these two opposing reactions to stress are not caused by genetic differences between cells.Lastly, Mizrachi et al. showed that how diatoms acclimate to stress depends on the amount of light they are exposed to. When in the dark, all cells became sensitive to oxidative stress, without forming distinct groups. But, when exposed to strong light that mimics the ocean surface, cells formed distinct groups within the population. This suggests that light regulates how susceptible these microscopic algae are to environmental stress.The different responses within a population may serve as a “bet-hedging” strategy, enabling at least some of the cells to survive unpredicted stressful conditions. The next challenge will be to find out whether algae growing in the oceans also use the same strategy and investigate what impact this has on diatom blooms.