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Plant roots play a dominant role in shaping the rhizosphere, the environment in which interaction with diverse microorganisms occurs. Tracking the dynamics of root–microbe interactions at high spatial resolution is currently limited because of methodological intricacy. Here, we describe a microfluidics-based approach enabling direct imaging of root–bacteria interactions in real time. The microfluidic device, which we termed tracking root interactions system (TRIS), consists of nine independent chambers that can be monitored in parallel. The principal assay reported here monitors behavior of fluorescently labeled Bacillus subtilis as it colonizes the root of Arabidopsis thaliana within the TRIS device. Our results show a distinct chemotactic behavior of B. subtilis toward a particular root segment, which we identify as the root elongation zone, followed by rapid colonization of that same segment over the first 6 h of root–bacteria interaction. Using dual inoculation experiments, we further show active exclusion of Escherichia coli cells from the root surface after B. subtilis colonization, suggesting a possible protection mechanism against root pathogens. Furthermore, we assembled a double-channel TRIS device that allows simultaneous tracking of two root systems in one chamber and performed real-time monitoring of bacterial preference between WT and mutant root genotypes. Thus, the TRIS microfluidics device provides unique insights into the microscale microbial ecology of the complex root microenvironment and is, therefore, likely to enhance the current rate of discoveries in this momentous field of research.

Coral reefs, and the unique ecosystems they support, are facing severe threats by human activities and climate change. Our understanding of these threats is hampered by the lack of robust approaches for studying the micro-scale interactions between corals and their environment. Here we present an experimental platform, coral-on-a-chip, combining micropropagation and microfluidics to allow direct microscopic study of live coral polyps. The small and transparent coral micropropagates are ideally suited for live-imaging microscopy, while the microfluidic platform facilitates long-term visualization under controlled environmental conditions. We demonstrate the usefulness of this approach by imaging coral micropropagates at previously unattainable spatio-temporal resolutions, providing new insights into several micro-scale processes including coral calcification, coral–pathogen interaction and the loss of algal symbionts (coral bleaching). Coral-on-a-chip thus provides a powerful method for studying coral physiology in vivo at the micro-scale, opening new vistas in coral biology. Studies of coral in laboratory settings are limited by a lack of reliable methods for manipulation of the coral microenvironment and monitoring of coral processes in vitro . Here the authors develop coral-on-a-chip, a microfluidic platform to enable study of coral processes at single-cell resolution.

Diseases are an emerging threat to ocean ecosystems. Coral reefs, in particular, are experiencing a worldwide decline because of disease and bleaching, which have been exacerbated by rising seawater temperatures. Yet, the ecological mechanisms behind most coral diseases remain unidentified. Here, we demonstrate that a coral pathogen, Vibrio coralliilyticus , uses chemotaxis and chemokinesis to target the mucus of its coral host, Pocillopora damicornis . A primary driver of this response is the host metabolite dimethylsulfoniopropionate (DMSP), a key element in the global sulfur cycle and a potent foraging cue throughout the marine food web. Coral mucus is rich in DMSP, and we found that DMSP alone elicits chemotactic responses of comparable intensity to whole mucus. Furthermore, in heat-stressed coral fragments, DMSP concentrations increased fivefold and the pathogen’s chemotactic response was correspondingly enhanced. Intriguingly, despite being a rich source of carbon and sulfur, DMSP is not metabolized by the pathogen, suggesting that it is used purely as an infochemical for host location. These results reveal a new role for DMSP in coral disease, demonstrate the importance of chemical signaling and swimming behavior in the recruitment of pathogens to corals and highlight the impact of increased seawater temperatures on disease pathways.

Changes in the microbial community composition of a full-scale membrane bioreactor treating industrial wastewater were studied over a period of 462 days using a series of 16S rRNA gene clone libraries. Frequent changes in the relative abundance of specific taxonomic groups were observed, which could not be explained by changes in the reactor's conditions or wastewater composition. Phage activity was proposed to drive some of the observed changes. Bacterial hosts were isolated from a biomass sample obtained towards the end of the study period, and specific phage counts were carried out for some of the isolated hosts using stored frozen biomass samples as the phage inocula. Plaque-forming unit concentrations were shown to change frequently over the study period, in correlation with changes in the relative abundance of taxonomic groups closely related by 16S rRNA gene sequence to the isolated strains. Quantitative PCR was used to verify changes in the abundance of a taxonomic group closely related to one of the isolated hosts, showing good agreement with the changes in relative abundance in the clone libraries of that group. The emerging pattern was consistent with the ‘killing the winner’ hypothesis, although alternative interaction mechanisms could not be ruled out. This is the first time that phage–host interactions in a complex microbial community are demonstrated over an extended period, and possibly the first in situ demonstration of ‘killing the winner’ stochastic behavior.

For some pathogenic fungi, sensing surface topography is part of their infection strategy. Their directional growth and transformation to a new developmental stage is influenced by contact with topographic features, which is referred to as thigmo-response, the exact functionality of which is not fully understood. Research on thigmo-responses is often performed on biomimetically patterned surfaces (BPS). Polydimethylsiloxane (PDMS) is especially suitable for fabrication of BPS. Here, we used synthetic BPS surfaces, mimicking tomato leaf surface, made from PDMS with the pathogenic fungus Botrytis cinerea to study the influence of structural features of the leaf surface on the fungus behavior. As a control, a PDMS surface without microstructure was fabricated to maintain the same chemical properties. Pre-penetration processes of B. cinerea , including the distribution of conidia on the surface, germination, and germ tube growth were observed on both leaf-patterned and flat PDMS. Microstructure affected the location of immediate attachment of conidia. Additionally, the microstructure of the plant host stimulated the development of germ tube in B. cinerea , at a higher rate than that observed on flat surface, suggesting that microstructure plays a role in fungus attachment and development.

Vibrio vulnificus is an emergent marine pathogen and is the cause of a deadly septicemia. However, the genetic factors that differentiate its clinical and environmental strains and its several biotypes remain mostly enigmatic. In this work, we investigated the underlying genomic properties and population dynamics of the V. vulnificus species to elucidate the traits that make these strains emerge as a human pathogen. The acquisition of different ecological determinants could have allowed the development of highly divergent clusters with different lifestyles within the same environment. However, we identified strains from both clusters in the mucosa of aquaculture species, indicating that manmade niches are bringing strains from the two clusters together, posing a potential risk of recombination and of emergence of novel variants. We propose a new evolutionary model that provides a perspective that could be broadly applicable to other pathogenic vibrios and facultative bacterial pathogens to pursue strategies to prevent their infections. Vibrio vulnificus , an opportunistic pathogen, is the causative agent of a life-threatening septicemia and a rising problem for aquaculture worldwide. The genetic factors that differentiate its clinical and environmental strains remain enigmatic. Furthermore, clinical strains have emerged from every clade of V. vulnificus . In this work, we investigated the underlying genomic properties and population dynamics of the V. vulnificus species from an evolutionary and ecological point of view. Genome comparisons and bioinformatic analyses of 113  V. vulnificus isolates indicate that the population of V. vulnificus is made up of four different clusters. We found evidence that recombination and gene flow between the two largest clusters (cluster 1 [C1] and C2) have drastically decreased to the point where they are diverging independently. Pangenome and phenotypic analyses showed two markedly different lifestyles for these two clusters, indicating commensal (C2) and bloomer (C1) ecotypes, with differences in carbohydrate utilization, defense systems, and chemotaxis, among other characteristics. Nonetheless, we identified frequent intra- and interspecies exchange of mobile genetic elements (e.g., antibiotic resistance plasmids, novel “chromids,” or two different and concurrent type VI secretion systems) that provide high levels of genetic diversity in the population. Surprisingly, we identified strains from both clusters in the mucosa of aquaculture species, indicating that manmade niches are bringing strains from the two clusters together. We propose an evolutionary model of V. vulnificus that could be broadly applicable to other pathogenic vibrios and facultative bacterial pathogens to pursue strategies to prevent their infections and emergence. IMPORTANCE Vibrio vulnificus is an emergent marine pathogen and is the cause of a deadly septicemia. However, the genetic factors that differentiate its clinical and environmental strains and its several biotypes remain mostly enigmatic. In this work, we investigated the underlying genomic properties and population dynamics of the V. vulnificus species to elucidate the traits that make these strains emerge as a human pathogen. The acquisition of different ecological determinants could have allowed the development of highly divergent clusters with different lifestyles within the same environment. However, we identified strains from both clusters in the mucosa of aquaculture species, indicating that manmade niches are bringing strains from the two clusters together, posing a potential risk of recombination and of emergence of novel variants. We propose a new evolutionary model that provides a perspective that could be broadly applicable to other pathogenic vibrios and facultative bacterial pathogens to pursue strategies to prevent their infections.

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.

Significance The fitness of corals and their ability to form large reefs hinge on their capacity to exchange oxygen and nutrients with their environment. Lacking gills or other ventilating organs, corals have been commonly assumed to depend entirely on ambient flow to overcome the mass transport limitations associated with molecular diffusion. Here, we show that corals are not enslaved to ambient flow but instead, can actively enhance mass transport by producing intense vortical flows with their epidermal cilia. By vigorously stirring the water immediately adjacent to their surface, this active process allows corals to increase mass transport and thus, can be a fundamental survival mechanism in regions or at times of weak ambient flow.

The complex structures and morphologies of coral skeletons make it difficult to study the construction of individual skeleton components. This is especially true regarding micro-structures deposited during different stages of skeletogenesis. As such structures serve as the basis for subsequent skeleton development, they are often obscured by additional skeletal deposition and growth. Recently we described a new model system, coral-on-a-chip, for studying coral biology and skeletogenesis. This system utilizes micropropagates of the Indo-Pacific coral Pocillopora damicornis maintained within a microfluidic environment, allowing us to follow early stages of skeletogenesis over time and under controlled environmental conditions. Our findings reveal that, following settlement onto glass slides, the micropropagates initially form a thin, almost two dimensional skeleton, which subsequently develops into a robust three-dimensional form with features resembling those of the mother colony. Studying the early stages of skeleton accretion in our micropropagates using high-resolution scanning electron microscopy and Energy Dispersive X-ray Spectroscopy (EDS) revealed a magnesium-rich layer deposited directly on the glass surface. This layer, which has a typical Mg/Ca ratio of 1.43±0.78, forms a dense lattice with a typical pore size of 100 nm. Microscopic observations indicate that this lattice serves as the basis for subsequent growth of fibrous aragonite. Examination of the underside of a skeleton from a small P. damicornis colony growing on a glass surface revealed a similar high-magnesium lattice at the interface between the glass and aragonitic skeleton in association with fibrous aragonite deposition. These observations suggest a role for this magnesium-rich lattice in the deposition of the fibrous aragonite forming the bulk of the coral skeleton.

Universal 16S rRNA gene primers (8F and 518R) bearing inosine substitutions at either the 3′-ultimate or the 3′-ultimate and penultimate base positions were exploited for the first time to study the bacterial community associated with coral polymicrobial Black Band Disease (BBD). Inosine-modified universal primer pairs display some shifting in the composition of 16S rRNA gene libraries, as well as expanding the observed diversity of a BBD bacterial community at the family/class level. Possible explanations for the observed shifts are discussed. These results thus point to the need for adopting multiple approaches in designing 16S rRNA universal primers for PCR amplification and subsequent construction of 16S rRNA gene libraries or pyrosequencing in the exploration of complex microbial communities.