Shapiro, O. H. | bioRxiv Science

bioRxiv2018

Phenotypic variability in chloroplast redox state predicts cell fate in a marine diatom

Diatoms are photosynthetic microorganisms of great ecological and biogeochemical importance, forming vast blooms in diverse aquatic ecosystems. Current understanding of phytoplankton acclimation to stress is based on population-level analysis, masking cell-to-cell variability. Here we investigated heterogeneity within Phaeodactylum tricornutum populations in response to oxidative stress, which is induced by environmental stress conditions. We combined flow cytometry and a microfluidics system for live imaging to measure redox dynamics at the single-cell level using the roGFP sensor. Chloroplast-targeted roGFP exhibited a light-dependent, bi-stable oxidation pattern in response to H2O2, revealing distinct subpopulations of sensitive oxidized cells and resilient reduced cells. Subpopulation proportions depended on growth phase, linking the bi-stable phenotype to proliferation. Oxidation of chloroplast-targeted roGFP preceded commitment to cell death and was used as a novel cell fate predictor. We propose that light-dependent metabolic heterogeneity results in differential stress responses that regulate cell fate within diatom populations.

microbiology↗

bioRxiv2018

Microscale tracking of coral disease reveals timeline of infection and heterogeneity of polyp fate.

Coral disease is often studied at scales ranging from single colonies to the entire reef. This is particularly true for studies following disease progression through time. To gain a mechanistic understanding of key steps underlying infection dynamics, it is necessary to study disease progression, and host-pathogen interactions, at relevant microbial scales. Here we provide a dynamic view of the interaction between the model coral pathogen Vibrio coralliilyticus and its coral host Pocillopora damicornis at unprecedented spatial and temporal scales. This view is achieved using a novel microfluidics-based system specifically designed to allow microscopic study of coral infection in-vivo under controlled environmental conditions. Analysis of exudates continuously collected at the systems outflow, allows a detailed biochemical and microbial analyses coupled to the microscopic observations of the disease progression. The resulting multilayered dataset provides the most detailed description of a coral infection to-date, revealing distinct pathogenic processes as well as the defensive behavior of the coral host. We provide evidence that infection in this system occurs following ingestion of the pathogen, and may then progress through the gastrovascular system. We further show infection may spread when pathogens colonize lesions in the host tissue. Copious spewing of pathogen-laden mucus from the polyp mouths results in effective expulsion of the pathogen from the gastrovascular system, possibly serving as a first line of defense. A secondary defense mechanism entails the severing of calicoblastic connective tissues resulting in the controlled isolation of diseased polyps, or the survival of individual polyps within infected colonies. Further investigations of coral-pathogen interactions at these scales will help to elucidate the complex interactions underlying coral disease, as we as the versatile adaptive response of the coral ecosystems to fluctuating environments.

microbiology↗