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Phytoplankton have species-specific responses toward electrogenerated oxidants, allowing high-throughput species analysis. Herein, a fluoro-electrochemical method is used to expose single Chlamydomonas concordia vegetative cells at different points within their life cycle to electro-generated oxidants from seawater. The resulting decay in fluorescence from chlorophyll-a is measured as a function of time and drops to zero for phytoplankton adjacent to the electrode over a period of a few seconds. The chlorophyll-a transient timescale allows mother cells, which are distinctively larger and require a larger quantity of oxidants, to be distinguished from either zoospores or “growing” cells, while all the cells show the same intrinsic susceptibility modulated only by the size of the phytoplankton. These observations are essential for the future automated characterization of the speciation of phytoplankton populations as they show that there is no need to manually identify the life cycle stage.

To broaden our understanding of pelagic ecosystem responses to environmental change, it is essential that we improve the spatio-temporal resolution of in situ monitoring of phytoplankton communities. A key challenge for existing methods is in classifying and quantifying cells within the nanophytoplankton size range (2-20µm). This is particularly difficult when there are similarities in morphology, making visual differentiation difficult for both trained taxonomists and machine learning based approaches. Here we present a rapid fluoro-electrochemical technique for classifying nanophytoplankton, and using a library of 52 diverse strains of nanophytoplankton we assess the accuracy of this technique based on two measurements at the individual level: charge required to reduce per cell chlorophyll a fluorescence by 50%, and cell radius. We demonstrate a high degree of accuracy overall (>90%) in categorising cells belonging to widely recognised key functional groups, however this is reduced when we consider the broader diversity of “nano-phytoflagellates”. Notably, we observe that some groups, for example calcifying Isochrysidales, have much greater resilience to electrochemically driven oxidative conditions relative to others of a similar size, making them more easily categorised by the technique. The findings of this study present a promising step forward in advancing our toolkit for monitoring phytoplankton communities. We highlight that, for improved categorisation accuracy, future iterations of the method can be enhanced by measuring additional predictor variables with minimal adjustments to the set-up. In doing so, we foresee this technique being highly applicable, and potentially invaluable, for in situ classification and enumeration of the nanophytoplankton size fraction.

Climate is partly controlled by atmospheric carbon dioxide levels. In turn atmospheric CO2 is substantially regulated by oceanic phytoplankton, specifically coccolithophores which biomineralize CO2 as CaCO3 in the form of tiny platelets which ultimately sink to the ocean floor. The scale of this biomineralization is massive; some 1015 g of CaCO3 are formed by phytoplankton per annum. This figure represents a tiny mass of CaCO3 per phytoplankton multiplied by a huge number of individual coccolithophores. For modelling the climate, it is essential to measure the biomineralized CaCO3 at single entity as well as bulk level. In this work we show that a simple light microscopy approach can, by monitoring the undersaturation driven dissolution of calcite in deionised (DI) water, quantify sub-nanograms of calcite mass of heterococcoliths secreted by marine phytoplankton. We show that the biogenic calcite found in coccoliths dissolves at a comparable mass-transport rate to that of inorganic particulate calcite in DI water. We further show that undersaturation driven dissolution is not suitable for quantifying coccolithophores due to competing osmotic pressure effects.

Schematic of fluoro-electrochemical microscopy. (a) Cartoon E. huxleyi is green under normal light, but (b) emits red fluorescence under UV. (c) Placed near an oxidizing electrode, its fluorescence fades and ultimately (d) “switches off”.

Multidisciplinary approaches can significantly advance our understanding of complex systems. For instance, gene co-expression networks align prior knowledge of biological systems with studies in graph theory, emphasising pairwise gene to gene interactions. In this paper, we extend these ideas, promoting hypergraphs as an investigative tool for studying multi-way interactions in gene expression data. Additional freedoms are achieved by representing individual genes with hyperedges, and simultaneous testing each gene against many features/vertices. Further gene/hyperedge interactions can be captured and explored using the line graph representations, a techniques that also reduces the complexity of dense hypergraphs. Such an approach provides access to graph centrality measures, which in turn identify salient features within a data set, for instance dominant or hub-like hyperedges leading to key knowledge on gene expression. The validity of this approach is established through the study of gene expression data for the plant speciesSenecio lautusand results will be interpreted within this biological setting.

Coccoliths are plates of biogenic calcium carbonate secreted by calcifying marine phytoplankton; annually these phytoplankton are responsible for exporting >1 billion tonnes (1015 g) of calcite to the deep ocean. Rapid and reliable methods for assessing the degree of calcification are technically challenging because the coccoliths are micron sized and contain picograms (pg) of calcite. Here we pioneer an opto‐eletrochemical acid titration of individual coccoliths which allows 3D reconstruction of each individual coccolith via in situ optical imaging enabling direct inference of the coccolith mass. Coccolith mass ranging from 2 to 400 pg are reported herein, evidencing both inter‐ and intra‐species variation over four different species. We foresee this scientific breakthrough, which is independent of knowledge regarding the species and calibration‐free, will allow continuous monitoring and reporting of the degree of coccolith calcification in the changing marine environment., Acid dissolution enables 3D image reconstruction of coccoliths, providing mass estimation in the range of 2 to 400 picograms.

Although, in principle, the Coulter Counter technique yields an absolute measure of particle volume, in practice, calibration is near-universally employed. For regularly shaped and non-biological samples, the use of latex beads for calibration can provide sufficient accuracy. However, this is not the case with particles encased in biogenically formed calcite. To date, there has been no effective route by which a Coulter Counter can be calibrated to enable the calcification of coccolithophores─single cells encrusted with biogenic calcite─to be quantified. Consequently, herein, we seek to answer the following question: to what extent can a Coulter Counter be used to provide accurate information regarding the calcite content of a single-species coccolithophore population? Through the development of a new calibration methodology, based on the measurement and dynamic tracking of the acid-driven calcite dissolution reaction, a route by which the cellular calcite content can be determined is presented. This new method allows, for the first time, a Coulter Counter to be used to yield an absolute measurement of the amount of calcite per cell.

The efficiency of carbon sequestration by the biological pump could decline in the coming decades because respiration tends to increase more with temperature than photosynthesis. Despite these differences in the short‐term temperature sensitivities of photosynthesis and respiration, it remains unknown whether the long‐term impacts of global warming on metabolic rates of phytoplankton can be modulated by evolutionary adaptation. We found that respiration was consistently more temperature dependent than photosynthesis across 18 diverse marine phytoplankton, resulting in universal declines in the rate of carbon fixation with short‐term increases in temperature. Long‐term experimental evolution under high temperature reversed the short‐term stimulation of metabolic rates, resulting in increased rates of carbon fixation. Our findings suggest that thermal adaptation may therefore have an ameliorating impact on the efficiency of phytoplankton as primary mediators of the biological carbon pump., The primary production of marine phytoplankton could decline with warming if rates of respiration increase more with rising temperature than photosynthesis. We found that respiration was consistently more temperature dependent than photosynthesis across 18 diverse marine phytoplankton taxa, resulting in universal declines in the rate of carbon fixation with short‐term increases in temperature. Whereas, by contrast, long‐term experimental evolution under high temperature acted to reverse the short‐term stimulation of metabolic rates, resulting in increased rates of carbon fixation.

The use of electro-generated oxidants in seawater facilitates the discrimination of different plankton groups via monitoring the decay in real time of their chlorophyll-a (chl-a) fluorescence signals following potentiostatic initiation of electrolysis in their vicinity (Yang, M. Chem. Sci.2019, 10(34), 7988−7993). In this paper, we explore the sensitivity of phytoplankton to different chemical species produced at various potentials in seawater. At low potentials, the oxidation of ca. millimolar bromide naturally present in seawater to hypobromous acid ‘switches-off’ the chl-a signal of individual Chlamydomonas concordia cells (green algae) located on the electrode surface within tens of seconds of the potential onset. At higher oxidative potentials, the oxidation of chloride and water produces oxidants (Cl2, OH, H2O2, etc.) that are also lethal to the plankton. To deconvolute the contributions to the response from the chemical identity of the oxidant and the amount of charge delivered to ‘titrate’ the individual living plankton using the loss of fluorescence as the ‘end point’, we introduce a ramped galvanostatic method. This approach enabled the controlled injection of charge applied to a bespoke electrochemical cell in which the plankton are immobilized on an electrode surface for rapid and sensitive measurement. It is shown that the number of moles (charge) of oxidants required to react leading to chl-a switch-off is independent of the chemical identity of the electro-generated oxidant(s) among hypobromous acid, chlorine, or water-derived oxidants. Comparative experiments between C. concordia and Emiliania huxleyi (where the latter are encapsulated by extracellular plates of calcium carbonate) show that significantly different amounts of absolute charge (moles of electro-generated oxidants) are required in each case to ‘switch-off’ the chl-a signal. The method provides the basis for a tool that could distinguish between different plankton cells within ca. 2 min including the setup time.

International audience; The 21st century has seen an acceleration of anthropogenic climate change and biodiversity loss, with both stressors deemed to affect ecosystem functioning. However, we know little about the interactive effects of both stressors and in particular about the interaction of increased climatic variability and biodiversity loss on ecosystem functioning. This should be remedied because larger climatic variability is one of the main features of climate change. Here, we demonstrated that temperature fluctuations led to changes in the importance of biodiversity for ecosystem functioning. We used microcosm communities of different phytoplankton species richness and exposed them to a constant, mild, and severe temperature-fluctuating environment. Wider temperature fluctuations led to steeper biodiversity–ecosystem functioning slopes, meaning that species loss had a stronger negative effect on ecosystem functioning in more fluctuating environments. For severe temperature fluctuations, the slope increased through time due to a decrease of the productivity of species-poor communities over time. We developed a theoretical competition model to better understand our experimental results and showed that larger differences in thermal tolerances across species led to steeper biodiversity–ecosystem functioning slopes. Species-rich communities maintained their ecosystem functioning with increased fluctuation as they contained species able to resist the thermally fluctuating environments, while this was on average not the case in species-poor communities. Our results highlight the importance of biodiversity for maintaining ecosystem functions and services in the context of increased climatic variability under climate change.

International audience; Rising sea surface temperatures are expected to lead to the loss of phytoplankton biodiversity. However, we currently understand very little about the interactions between warming, loss of phy-toplankton diversity and its impact on the oceans' primary production. We experimentally manipulated the species richness of marine phytoplankton communities under a range of warming scenarios, and found that ecosystem production declined more abruptly with species loss in communities exposed to higher temperatures. Species contributing positively to ecosystem production in the warmed treatments were those that had the highest optimal temperatures for photosynthe-sis, implying that the synergistic impacts of warming and biodiversity loss on ecosystem functioning were mediated by thermal trait variability. As species were lost from the communities, the probability of taxa remaining that could tolerate warming diminished, resulting in abrupt declines in ecosystem production. Our results highlight the potential for synergistic effects of warming and biodiversity loss on marine primary production.

Marine phytoplankton are responsible for over 45% of annual global net primary production. Ocean warming is expected to drive massive reorganisation of phytoplankton communities, resulting in pole-ward range shifts and sharp declines in species diversity, particularly in the tropics. The impacts of warming on phytoplankton species depend critically on their physiological sensitivity to temperature change, characterised by thermal tolerance curves. Local extinctions arise when temperatures exceed species’ thermal tolerance limits. The mechanisms that determine the characteristics of thermal tolerance curves (e.g. optimal and maximal temperatures) and their variability among the broad physiological diversity of marine phytoplankton are however poorly understood. Here we show that differences in the temperature responses of photosynthesis and respiration establish physiological trade-offs that constrain the thermal tolerance of 18 species of marine phytoplankton, spanning cyanobacteria as well as the red and green super-families. Across all species we found that rates of respiration were more sensitive to increasing temperature and typically had higher optimal temperatures than photosynthesis. Consequently, the fraction of photosynthetic energy available for allocation to growth (carbon-use efficiency) declined exponentially with rising temperatures with a sensitivity that was invariant among the 18 species. Furthermore, the optimal temperature of growth was generally lower than that of photosynthesis and as a result, supra-optimal declines in growth rate were associated with temperature ranges where the carbon-use efficiency exhibited accelerated declines. These highly conserved patterns demonstrate that the limits of thermal tolerance in marine phytoplankton are underpinned by common metabolic constraints linked to the differential temperature responses of photosynthesis and respiration.Significance StatementThe impacts of warming on marine phytoplankton depend on their sensitivity to rising temperatures, yet there is currently limited understanding of the mechanisms that limit thermal tolerance among the diversity of marine phytoplankton. Using a comparative study on the dominant, ecologically important lineages of marine phytoplankton – Bacillariophyceae, Dinophyceae, Cyanophyceae, Prasinophyceae, Prymnesiophyceae – we show that rates of respiration are consistently more sensitive to increasing temperature than photosynthesis. Consequently, the fraction of photosynthetic energy available for growth declines with rising temperatures with a sensitivity that is invariant among species. Our results suggest that declines in phytoplankton performance at high temperatures are driven by universal metabolic constrains linked to rising respiratory costs eventually exceeding the supply of reduced carbon from photosynthesis.

Phytoplankton photosynthesis is a critical flux in the carbon cycle, accounting for approximately 40% of the carbon dioxide fixed globally on an annual basis and fuelling the productivity of aquatic food webs. However, rapid evolutionary responses of phytoplankton to warming remain largely unexplored, particularly outside the laboratory, where multiple selection pressures can modify adaptation to environmental change. Here, we use a decade-long experiment in outdoor mesocosms to investigate mechanisms of adaptation to warming (+4 °C above ambient temperature) in the green alga Chlamydomonas reinhardtii, in naturally assembled communities. Isolates from warmed mesocosms had higher optimal growth temperatures than their counterparts from ambient treatments. Consequently, warm-adapted isolates were stronger competitors at elevated temperature and experienced a decline in competitive fitness in ambient conditions, indicating adaptation to local thermal regimes. Higher competitive fitness in the warmed isolates was linked to greater photosynthetic capacity and reduced susceptibility to photoinhibition. These findings suggest that adaptive responses to warming in phytoplankton could help to mitigate projected declines in aquatic net primary production by increasing rates of cellular net photosynthesis. Isolates of the green alga Chlamydomonas reinhardtii adapted to a decade of chronic warming in outdoor mesocosms have higher competitive fitness than ambient controls, due to greater photosynthetic capacity and reduced susceptibility to photoinhibition

A number of articles and empirical studies over the past decade suggest a relationship between the criminal law’s reputation for being just—its “moral credibility”—and its ability to gain society’s deference and compliance through a variety of mechanisms that enhance the system’s crime-control effectiveness. This has led to proposals for criminal liability and punishment rules to reflect lay intuitions of justice—“empirical desert”—as a means of enhancing the system’s moral credibility. In a recent article, Christopher Slobogin and Lauren Brinkley-Rubinstein (SBR) report seven sets of studies that, in their view, undermine these claims about empirical desert and moral credibility. Instead, say SBR, the studies support their own proposed distributive principle of “individual prevention.” As this article shows, however, SBR have it wrong on both counts: not only do their studies actually confirm the crime-control power of empirical desert, but they provide no support for their own principle of individual prevention. Moreover, that principle, which focuses on an offender’s dangerousness rather than his perceived desert, is erroneously described by SBR as “a sort of limiting retributivism.” In reality, what SBR propose is a system based on dangerousness, where detention decisions are made at the back end by experts. Such an approach promotes the worst of the failed policies of the 1960s, and conflicts with the modern trend of encouraging more community involvement in criminal punishment, not less.

A number of articles and empirical studies over the past decade, most by Paul Robinson and co-authors, suggest a relationship between the criminal law’s reputation for being just — its "moral credibility" — and its ability to gain society’s deference and compliance through a variety of mechanisms that enhance its crime-control effectiveness. This has led to proposals to have criminal liability and punishment rules reflect lay intuitions of justice — "empirical desert" — as a means of enhancing the system's moral credibility. In a recent article, Christopher Slobogin and Lauren Brinkley-Rubinstein (SBR) report seven sets of studies that, they argue, undermine these claims about empirical desert and moral credibility and instead support their own proposed distributive principle of "individual prevention." As this article shows, however, SBR have it essentially backwards: not only do their studies actually confirm the crime-control power of empirical desert, but they provide no support for their own principle of individual prevention. Moreover, that principle, which focuses on an offender’s dangerousness rather than his perceived desert, is erroneously described by SBR as "a sort of limiting retributivism." In reality, what SBR propose is a system based on dangerousness, where detention decisions are made at the back-end by experts. Such an approach promotes the worst of the failed policies of the 1960s, and conflicts with the modern trend of encouraging more community involvement in criminal punishment, not less.