bioRxiv Science⌕ Search

Biology subjects

Poulain, A. J.

Publications and source records attributed to Poulain, A. J..

6 recordsLinked to original sources

Glacial meltwater drives gene-specific diversification of metal resistance genes in High Arctic soil microbiomes

Climate warming accelerates glacial meltwater delivery to Arctic lakes, mobilizing metals from thawing catchments and reshaping the selective landscape experienced by resident microbes. Whether these gradients leave detectable evolutionary signatures in environmental resistance genes remains unclear. We investigated four metal resistance genes (merA, arsC, cadA, and chrR) in metagenomic datasets from Lake Hazen (Nunavut, Canada), the largest High Arctic freshwater lake, sampled across a natural hydrological gradient of Control, Low-runoff, and High-runoff regimes. Using a space-for-time design, we combined population-genetic and codon-based approaches to quantify diversity and selection, including nucleotide diversity, Tajimas D, nonsynonymous/synonymous diversity ratios, McDonald-Kreitman tests, site-level episodic selection, and Bayesian coalescent and structured-coalescent inference of gene-pool diversity and exchange. This multi-pronged approach allowed us to distinguish gene-specific evolutionary responses from broader demographic effects. We found marked heterogeneity among genes: merA showed increasing diversity and consistent evidence of adaptive evolution along the runoff gradient; cadA displayed the strongest adaptive signal under low runoff, with selective constraint patterns that varied across regimes; chrR exhibited the clearest signature of episodic positive selection, with gene-wide selection and adaptive substitutions concentrated in the high-runoff regime where chromium concentrations were greatest; while arsC remained largely consistent with neutral evolution across regimes. Together, these results show that climate-driven metal mobilization is associated with gene- and regime-specific diversification of microbial metal resistance gene pools in Arctic soil microbiomes. Differences in metal speciation (with arsenic and chromium occurring as redox-sensitive oxyanions, and mercury, cadmium and zinc as divalent cations) may contribute to the contrasting evolutionary trajectories observed across genes, although in-situ speciation was not assessed in this dataset. Our findings highlight environmental resistance genes as sensitive indicators of changing biogeochemical conditions in rapidly warming polar ecosystems.

microbiology↗

Methane Cycling Microbes are Important Predictors of Methylmercury Accumulation in Rice Paddies

Microbial production of methylmercury from inorganic mercury in rice paddies poses health risks to consumers of this essential dietary staple. Although mercury-methylating communities are well characterized, the microbial guilds contributing to methylmercury accumulation in rice paddies remain unclear. Here, we collected paddy soils across a mercury concentration gradient throughout the rice growing season to identify microbial and environmental factors influencing methylmercury dynamics. We show that hgcA gene abundance, the key gene required for methylation, was not a significant predictor of methylmercury concentration in paddy soils. We also show that merB gene abundance correlated with methylmercury in mercury-polluted rhizosphere samples. Methane cycling genes were actively expressed, and their beta-diversity was significantly associated with methylmercury levels. Methanogen abundance correlated with higher methylmercury under elevated total mercury concentrations. Analysis of the methanotroph-associated mbnT gene, implicated in demethylation, revealed an unexpected positive correlation with methylmercury. Multiple regression and machine learning models converged on mercury bioavailability and methanogen/methanotroph abundances as key predictors of methylmercury, with methanogen-associated hgcA gene abundance and methanogen-methanotroph interactions highlighted under flooded, low-redox conditions. These findings suggest that methane-cycling microbes play key roles in methylmercury cycling dynamics and point to management strategies that could simultaneously mitigate mercury pollution and greenhouse gas emissions. ImportanceMethylmercury is a microbially-derived neurotoxin that accumulates in rice, which is a global food staple. Predicting mercurys fate in rice paddies is challenging because of the interplay between microbes responsible for methylmercury cycling and variables that control mercury availability. Our study coupled genomic and geochemical measurements with machine learning to identify the key predictors of methylmercury accumulation in paddy soils. We demonstrate that methanogen and methanotroph abundance, and mercury bioavailability, are major predictors of methylmercury variability in paddies. We show that considering interactions between methane cycling guilds improves our capacity to predict methylmercury accumulation in soils compared to approaches that rely solely on mercury cycling genes. This work can inform remediation strategies for mercury in rice paddies but also wetlands and permafrost where methane and mercury cycling are tightly coupled. Such strategies could provide a solution to simultaneously mitigate methylmercury exposure and reduce greenhouse gas emissions amid global environmental change.

microbiology↗

The High Arctic is dominated by uncharacterized, genetically highly diverse bacteriophages

While the Earths virosphere is estimated to be in the range of 1031 viral particles, the vast majority of its diversity has yet to be discovered. In recent years, metagenomics has rapidly allowed the identification of viruses, from microenvironments to extreme environments like the High Arctic. However, the High Arctic virome is largely composed of viral sequences that have few matches to classified viruses in existing databases. Here, to bypass limitations posed by similarity-based strategies, we resorted to a paired metagenomic (DNA, n = 8) and metatranscriptomic (RNA, n = 6) approach that placed viral genes found in Lake Hazen, the worlds largest High Arctic lake, in a phylogenetic context with known viruses. We show that High Arctic viruses have undergone unique evolutionary processes characterized by high evolutionary rates, making them distinct from and more diverse than known viruses.

evolutionary biology↗

Viral spillover risk in High Arctic increases with melting glaciers

The host spectrum of viruses is quite diverse, as they can sustainedly infect a few species to several phyla. When confronted with a new host, a virus may even infect it and transmit sustainably in this new host, a process called "viral spillover." However, the risk of such events is difficult to quantify. As climate change is rapidly transforming environments, it is becoming critical to quantify the potential for spillovers. To address this issue, we resorted to a metagenomics approach and focused on two environments, soil and lake sediments from Lake Hazen, the largest High Arctic freshwater lake in the world. We used DNA and RNA sequencing to reconstruct the lakes virosphere in both its sediments and soils, as well as its range of eukaryotic hosts. We then estimated the spillover risk by measuring the congruence between the viral and the eukaryotic host phylogenetic trees, and show that spillover risk is higher in lake sediments than in soil, and increases with runoff from glacier melt, a proxy for climate change. Should climate change also shift species range of potential viral vectors and reservoirs northwards, the High Arctic could become fertile ground for emerging pandemics.

ecology↗

Golden Gate Assembly of Aerobic and Anaerobic Microbial Bioreporters

Microbial bioreporters provide direct insight into cellular processes by producing a quantifiable signal dictated by reporter gene expression. The core of a bioreporter is a genetic circuit in which a reporter gene (or operon) is fused to promoter and regulatory sequences that govern its expression. In this study, we develop a system for constructing novel Escherichia coli bioreporters based on Golden Gate assembly, a synthetic biology approach for the rapid and seamless fusion of DNA fragments. Gene circuits are generated by fusing promoter and reporter sequences encoding yellow fluorescent protein, mCherry, bacterial luciferase, and an anaerobically active flavin-based fluorescent protein. We address a barrier to the implementation of Golden Gate assembly by designing a series of compatible destination vectors that can accommodate the assemblies. We validate the approach by measuring the activity of constitutive bioreporters and mercury and arsenic biosensors in quantitative exposure assays. We also demonstrate anaerobic quantification of mercury and arsenic in biosensors that produce flavin-based fluorescent protein, highlighting the expanding range of redox conditions that can be examined by microbial bioreporters. IMPORTANCEMicrobial bioreporters are versatile genetic tools with wide-ranging applications, particularly in the field of environmental toxicology. For example, biosensors that produce a signal output in the presence of a specific analyte offer less costly alternatives to analytical methods for the detection of environmental toxins such as mercury and arsenic. Biosensors of specific toxins can also be used to test hypotheses regarding mechanisms of uptake, toxicity, and biotransformation. In this study, we develop an assembly platform that uses a synthetic biology technique to streamline construction of novel Escherichia coli bioreporters that produce fluorescent or luminescent signals. We validate the approach by synthesizing and testing an array of bioreporters, including arsenic and mercury biosensors, that produce signal outputs in environments ranging from aerobic to highly reduced anaerobic growth conditions.

molecular biology↗

Common physiological processes control mercury reduction during photosynthesis and fermentation

Mercury (Hg) is a global pollutant and potent neurotoxin that bioaccumulates in food webs as monomethylmercury (MeHg). The production of MeHg is driven by anaerobic and Hg redox cycling pathways such as Hg reduction, which control the availability of Hg to methylators. Anaerobes play an important role in Hg reduction in methylation hotspots, yet their contributions remain underappreciated due to how challenging these pathways are to study in the absence of dedicated genetic targets and low levels of Hg0 in anoxic environments. In this study we used Hg stable isotope fractionation to explore Hg reduction during anoxygenic photosynthesis and fermentation in the model anaerobe Heliobacterium modesticaldum Ice1. We show that cells preferentially reduce lighter Hg isotopes in both metabolisms leading to mass-dependent fractionation, but mass-independent fractionation commonly induced by UV-visible light is absent. We show that isotope fractionation is affected by the interplay between pathways controlling Hg recruitment, accessibility, and availability alongside metabolic redox reactions. The combined contributions of these processes lead to isotopic enrichment during anoxygenic photosynthesis that is in between the values reported for anaerobic respiratory microbial Hg reduction and abiotic photoreduction. Isotope enrichment during fermentation is closer to what has been observed in aerobic bacteria that reduce Hg through dedicated detoxification pathways. These results demonstrate that common controls exist at the atomic level for Hg reduction during photosynthesis and fermentation in H. modesticaldum. Our work suggests that similar controls likely underpin diverse microbe-mediated Hg transformations that affect Hgs fate in oxic and anoxic habitats. IMPORTANCEAnaerobic and photosynthetic bacteria that reduce mercury affect mercury delivery to microbes in methylation sites that drive bioaccumulation in food webs. Anaerobic mercury reduction pathways remain underappreciated in the current view of the global mercury cycle because they are challenging to study, bearing no dedicated genetic targets to establish physiological mechanisms. In this study we used stable isotopes to show that common physiological processes control mercury reduction during photosynthesis and fermentation in the model anaerobe Heliobacterium modesticaldum Ice1. The sensitivity of isotope analyses highlighted the subtle contribution of mercury uptake towards the isotope signature associated with anaerobic mercury reduction. When considered alongside the isotope signatures associated with microbial pathways for which genetic determinants have been identified, our findings underscore the narrow range of isotope enrichment that is characteristic of microbial mercury transformations. This suggests that there exist common atomic-level controls for biological mercury transformations across a broad range of geochemical conditions.

microbiology↗