Microbial Dormancy Research Articles

Microbial Dormancy Supports Multi-Species Coexistence Under Resource Fluctuations.

The ability for microbes to enter dormant states is adaptive under resource fluctuations and has been linked to the maintenance of diversity. Nevertheless, the mechanism by which microbial dormancy gives rise to the density-dependent feedbacks required for stable coexistence under resource fluctuations is not well understood. Via analysis of consumer-resource models, we show that the stable coexistence of dormancy and non-dormancy strategists is a consequence of the former benefiting more from resource fluctuations while simultaneously reducing overall resource variability, which sets up the requisite negative frequency dependence. Moreover, we find that dormants can coexist alongside gleaner and opportunist strategies in a competitive-exclusion-defying case of three species coexistence on a single resource. This multi-species coexistence is typically characterised by non-simple assembly rules that cannot be predicted from pairwise competition outcomes. The diversity maintained via this three-way trade-off represents a novel phenomenon that is ripe for further theoretical and empirical inquiry.

The neglected roles of adjacent natural ecosystems in maintaining bacterial diversity in agroecosystems.

A central aim of community ecology is to understand how local species diversity is shaped. Agricultural activities are reshaping and filtering soil biodiversity and communities; however, ecological processes that structure agricultural communities have often overlooked the role of the regional species pool, mainly owing to the lack of large datasets across several regions. Here, we conducted a soil survey of 941 plots of agricultural and adjacent natural ecosystems (e.g., forest, wetland, grassland, and desert) in 38 regions across diverse climatic and soil gradients to evaluate whether the regional species pool of soil microbes from adjacent natural ecosystems is important in shaping agricultural soil microbial diversity and completeness. Using a framework of multiscales community assembly, we revealed that the regional species pool was an important predictor of agricultural bacterial diversity and explained a unique variation that cannot be predicted by historical legacy, large-scale environmental factors, and local community assembly processes. Moreover, the species pool effects were associated with microbial dormancy potential, where taxa with higher dormancy potential exhibited stronger species pool effects. Bacterial diversity in regions with higher agricultural intensity was more influenced by species pool effects than that in regions with low intensity, indicating that the maintenance of agricultural biodiversity in high-intensity regions strongly depends on species present in the surrounding landscape. Models for community completeness indicated the positive effect of regional species pool, further implying the community unsaturation and increased potential in bacterial diversity of agricultural ecosystems. Overall, our study reveals the indubitable role of regional species pool from adjacent natural ecosystems in predicting bacterial diversity, which has useful implication for biodiversity management and conservation in agricultural systems.

Ribosomal RNA gene operon copy number, a functional trait indicating the hydrocarbon degradation level of bacterial communities

The lack of universal indicators for predicting microbial biodegradation potential and assessing remediation effects limits the generalization of bioremediation. The community-level ribosomal RNA gene operon (rrn) copy number, an important functional trait, has the potential to serve as a key indicator of the bioremediation of organic pollutants. A meta-analysis based on 1275 samples from 26 hydrocarbon-related studies revealed a positive relationship between the microbial hydrocarbon biodegradation level and the community-level rrn copy number in soil, seawater and culture. Subsequently, a microcosm experiment was performed to decipher the community-level rrn copy number response mechanism during total petroleum hydrocarbon (TPH) biodegradation. The treatment combining straw with resuscitation-promoting factor (Rpf) exhibited the highest community-level rrn copy number and the most effective biodegradation compared with other treatments, and the initial TPH content (20,000 mg kg-1) was reduced by 67.67% after 77 days of incubation. TPH biodegradation rate was positively correlated with the average community-level rrn copy number (p = 0.001, R2 = 0.5781). Both meta and community analyses showed that rrn copy number may reflect the potential of hydrocarbon degradation and microbial dormancy. Our findings provide insight into the applicability of the community-level rrn copy number to assess bacterial biodegradation for pollution remediation.

Extended lag phase indicates the dormancy of biphenyl degrading Rhodococcus biphenylivorans TG9 under heat stress

Microbial remediation is a green and sustainable technology, but harsh environmental conditions could lead to microbial dormancy, such as entering a viable but non-culturable (VBNC) state. However, the evidence of VBNC is controversial and limited. In this study, heat stress (60 °C), one of the leading challenges for mesophilic degrading bacteria, was mimicked to investigate the physiological response of Rhodococcus biphenylivorans TG9. After 2 h of heat stress, the culturable TG9 cell count decreased from 108 cells/mL to undetectable while the viable cell count was still 105 cells/mL. The biphenyl degradation efficiency of stressed TG9 dropped by 50% compared to that of cells at logarithmic phase. During heat stress, the respiratory activity of TG9 declined dramatically while the intracellular ATP level initially increased and then decreased. Notably, the corresponding indicators recovered when restored to 30 °C. These characteristics were in consistent with bacteria entering into VBNC state. Furthermore, fluorescence activated cell sorting together with single cell as seed culture detection verified the unculturability and viability of VBNC state of TG9 cells. Also, we found that single cells in VBNC state could resuscitate and regrowth with significantly extended lag phase (LP). Our results highlight the potential of TG9 for microbial remediation and hint LP duration as an indicator for survival state of bacteria.

Effects of Desiccation and Freezing on Microbial Ionizing Radiation Survivability: Considerations for Mars Sample Return.

Increasingly, national space agencies are expanding their goals to include Mars exploration with sample return. To better protect Earth and its biosphere from potential extraterrestrial sources of contamination, as set forth in the Outer Space Treaty of 1967, international efforts to develop planetary protection measures strive to understand the danger of cross-contamination processes in Mars sample return missions. We aim to better understand the impact of the martian surface on microbial dormancy and survivability. Radiation resistance of microbes is a key parameter in considering survivability of microbes over geologic times on the frigid, arid surface of Mars that is bombarded by solar and galactic cosmic radiation. We tested the influence of desiccation and freezing on the ionizing radiation survival of six model microorganisms: vegetative cells of two bacteria (Deinococcus radiodurans, Escherichia coli) and a strain of budding yeast (Saccharomyces cerevisiae); and vegetative cells and endospores of three Bacillus bacteria (B. subtilis, B. megaterium, B. thuringiensis). Desiccation and freezing greatly increased radiation survival of vegetative polyploid microorganisms when applied separately, and when combined, desiccation and freezing increased radiation survival even more so. Thus, the radiation survival threshold of polyploid D. radiodurans cells can be extended from the already high value of 25 kGy in liquid culture to an astonishing 140 kGy when the cells are both desiccated and frozen. However, such synergistic radioprotective effects of desiccation and freezing were not observed in monogenomic or digenomic Bacillus cells and endospores, which are generally sterilized by 12 kGy. This difference is associated with a critical requirement for survivability under radiation, that is, repair of genome damage caused by radiation. Deinococcus radiodurans and S. cerevisiae accumulate similarly high levels of the Mn antioxidants that are required for extreme radiation resistance, as do endospores, though they greatly exceed spores in radioresistance because they contain multiple identical genome copies, which in D. radiodurans are joined by persistent Holliday junctions. We estimate ionizing radiation survival limits of polyploid DNA-based life-forms to be hundreds of millions of years of background radiation while buried in the martian subsurface. Our findings imply that forward contamination of Mars will essentially be permanent, and backward contamination is a possibility if life ever existed on Mars.

Farm-scale differentiation of active microbial colonizers

Microbial movement is important for replenishing lost soil microbial biodiversity and driving plant root colonization, particularly in managed agricultural soils, where microbial diversity and composition can be disrupted. Despite abundant survey-type microbiome data in soils, which are obscured by legacy DNA and microbial dormancy, we do not know how active microbial pools are shaped by local soil properties, agricultural management, and at differing spatial scales. To determine how active microbial colonizers are shaped by spatial scale and environmental conditions, we deployed microbial traps (i.e. sterile soil enclosed by small pore membranes) containing two distinct soil types (forest; agricultural), in three neighboring locations, assessing colonization through 16S rRNA gene and fungal ITS amplicon sequencing. Location had a greater impact on fungal colonizers (R2 = 0.31 vs. 0.26), while the soil type within the microbial traps influenced bacterial colonizers more (R2 = 0.09 vs. 0.02). Bacterial colonizers showed greater colonization consistency (within-group similarity) among replicate communities. Relative to bacterial colonizers, fungal colonizers shared a greater compositional overlap to sequences from the surrounding local bulk soil (R2 = 0.08 vs. 0.29), suggesting that these groups respond to distinct environmental constraints and that their in-field management may differ. Understanding how environmental constraints and spatial scales impact microbial recolonization dynamics and community assembly are essential for identifying how soil management can be used to shape agricultural microbiomes.

Uncertainty quantification of the soil moisture response functions for microbial dormancy and resuscitation

Abiotic factors influence ecosystem functioning mainly through the biological mechanisms such as the shifts in microbial community structure and/or physiology. Sigmoidal shape soil moisture response functions (SMRFs) have been widely used to modify the dormancy and resuscitation rates in microbially explicit soil biogeochemical models. However, these SMRFs and their effects on microbial dormancy have not been well quantified with experimental data. Using observations from a three-year field experiment in subtropical forests, we conducted a comprehensive parameterization of typical sigmoidal SMRFs. We quantified uncertainties in the three parameters of these SMRFs: (i) the critical soil water potential (SWP) for microbes entering dormancy, φA2D=−0.46±0.03MPa (mean ± standard deviation); (ii) the ratio of the critical SWP for resuscitation to φA2D, τ=0.39±0.16; and (iii) the exponent in the SMRFs, ω=3.38±0.54. Our results show much higher uncertainty in the SMRF for microbial resuscitation than microbial dormancy, due to the parameter τ being included in microbial reactivation with larger uncertainty than that of φA2D and ω. In addition, the active microbes, rather than the total living microbes, were overwhelmingly controlled by soil moisture, leading to the stronger dependency of microbial respiration on soil moisture in the dry season than in the wet season. The SMRFs and their parameter values/uncertainties derived in this study could be directly incorporated into or used as a priori in ecosystem and Earth system models that account for microbial dormancy and resuscitation.

Resuscitation of the microbial seed bank alters plant-soil interactions.

While microorganisms are recognized for driving belowground processes that influence the productivity and fitness of plant populations, the vast majority of bacteria and fungi in soil belong to a seed bank consisting of dormant individuals. However, plant performance may be affected by microbial dormancy through its effects on the activity, abundance, and diversity of soil microorganisms. To test how microbial seed banks influence plant-soil interactions, we purified recombinant resuscitation promoting factor (Rpf), a bacterial protein that terminates dormancy. In a factorially designed experiment, we then applied the Rpf to soil containing field mustard (Brassica rapa), an agronomically important plant species. Plant biomass was ~33% lower in the Rpf treatment compared to plants grown with an unmanipulated microbial seed bank. In addition, Rpf reduced soil respiration, decreased bacterial abundance, and increased fungal abundance. These effects of Rpf on plant performance were accompanied by shifts in bacterial community composition, which may have diluted mutualists or resuscitated pathogens. Our findings suggest that changes in microbial seed banks may influence the magnitude and direction of plant-soil feedbacks in ways that affect above- and belowground biodiversity and function.

Microbial dormancy and its impacts on northern temperate and boreal terrestrial ecosystem carbon budget

Abstract. A large amount of soil carbon in northern temperate and boreal regions could be emitted as greenhouse gases in a warming future. However, lacking detailed microbial processes such as microbial dormancy in current biogeochemistry models might have biased the quantification of the regional carbon dynamics. Here the effect of microbial dormancy was incorporated into a biogeochemistry model to improve the quantification for the last century and this century. Compared with the previous model without considering the microbial dormancy, the new model estimated the regional soils stored 75.9 Pg more C in the terrestrial ecosystems during the last century and will store 50.4 and 125.2 Pg more C under the RCP8.5 and RCP2.6 scenarios, respectively, in this century. This study highlights the importance of the representation of microbial dormancy in earth system models to adequately quantify the carbon dynamics in the northern temperate and boreal natural terrestrial ecosystems.

Invasion and fixation of microbial dormancy traits under competitive pressure

Microbial dormancy is an evolutionary trait that has emerged independently at various positions across the tree of life. It describes the ability of a microorganism to switch to a metabolically inactive state that can withstand unfavourable conditions. However, maintaining such a trait requires additional resources that could otherwise be used to increase e.g. reproductive rates. In this paper, we aim for gaining a basic understanding under which conditions maintaining a seed bank of dormant individuals provides a “fitness advantage” when facing resource limitations and competition for resources among individuals (in an otherwise stable environment). In particular, we wish to understand when an individual with a “dormancy trait” can invade a resident population lacking this trait despite having a lower reproduction rate than the residents. To this end, we follow a stochastic individual-based approach employing birth-and-death processes, where dormancy is triggered by competitive pressure for resources. In the large-population limit, we identify a necessary and sufficient condition under which a complete invasion of mutants has a positive probability. Further, we explicitly determine the limiting probability of invasion and the asymptotic time to fixation of mutants in the case of a successful invasion. In the proofs, we observe the three classical phases of invasion dynamics in the guise of Coron et al. (2017, 2019).

Mimicking climate warming effects on Alaskan soil microbial communities via gradual temperature increase

Climate change can trigger shifts in community structure and may therefore pose a severe threat to soil microbial communities, especially in high northern latitudes such as the Arctic. Arctic soils are covered by snow and ice throughout most of the year. This insulation shields them from high temperature variability and low surface temperatures. If this protective layer thaws, these soils are predicted to warm up at 1.5x to 4x the rate of other terrestrial biomes. In this study, we sampled arctic soils from sites with different elevations in Alaska, incubated them for 5 months with a simulated, gradual or abrupt temperature increase of +5 °C, and compared bacterial and fungal community compositions after the incubation. We hypothesized that the microbial communities would not significantly change with a gradual temperature treatment, whereas an abrupt temperature increase would decrease microbial diversity and shift community composition. The only differences in community composition that we observed were, however, related to the two elevations. The abrupt and gradual temperature increase treatments did not change the microbial community composition as compared to the control indicating resistance of the microbial community to changes in temperature. This points to the potential importance of microbial dormancy and resting stages in the formation of a “buffer” against elevated temperatures. Microbial resting stages might heavily contribute to microbial biomass and thus drive the responsiveness of arctic ecosystems to climate change.

Modeling the processes of soil moisture in regulating microbial and carbon-nitrogen cycling

Soil carbon (C) and nitrogen (N) cycles and their complex responses to hydro-climatic forcing have gained increasing attention. While the temperature effects have been intensively studied, soil moisture response functions (SMRFs) are not well documented for various microbial and enzymatic processes due to the difficulties in directly measuring and differentiating the moisture effects on various processes. Here we extended our C-only Microbial-ENzyme Decomposition (MEND) model to the C-N coupled MEND model with flexible element stoichiometry. Our model calibration showed good agreement between simulated and observed C:N ratios in soil organic matter and microbial biomass, as well as the ammonium and nitrate concentrations. We show that the selection of SMRFs for specific biogeochemical processes could result in significant differences in model simulated microbial and C-N processes. In particular, it is essential to account for the soil moisture effects on microbial dormancy and resuscitation, as the changes in microbial physiology under favorable or stressful conditions will exert strong controls on soil C and N dynamics. We also advocate the utilization of dynamic (time-variant) data (e.g., litter input, N deposition, soil temperature and moisture), instead of time-invariant data, to drive model simulations and analyses. Dynamic forcing data (particularly dynamic soil moisture) better represent the real-world climate and environmental conditions, which could facilitate more realistic modeling and understanding of soil C and nutrient cycling in a changing world.

The seed bank coalescent with simultaneous switching

We introduce a new Wright-Fisher type model for seed banks incorporating “simultaneous switching”, which is motivated by recent work on microbial dormancy ([21], [28]). We show that the simultaneous switching mechanism leads to a new jump-diffusion limit for the scaled frequency processes, extending the classical Wright-Fisher and seed bank diffusion limits. We further establish a new dual coalescent structure with multiple activation and deactivation events of lineages. While this seems reminiscent of multiple merger events in general exchangeable coalescents, it actually leads to an entirely new class of coalescent processes with unique qualitative and quantitative behaviour. To illustrate this, we provide a novel kind of condition for coming down from infinity for these coalescents, applying a recent approach of Griffiths [12].

Microbial dormancy improves predictability of soil respiration at the seasonal time scale

Global warming, in combination with altered precipitation patterns, is accelerating global soil respiration, which could in turn accelerate climate change. The biological mechanisms through which soil carbon (C) responds to climate are not well understood, limiting our ability to predict future global soil respiration rates. As part of a climate manipulation experiment, we tested whether differences in soil heterotrophic respiration (RH) driven by season or climate treatment are linked to (1) relative abundances of microbes in active and dormant metabolic states, (2) net changes in microbial biomass and/or (3) changes in the relative abundances of microbial groups with different C-use strategies. We used a flow-cytometric single-cell metabolic assay to quantify the abundance of active and dormant microbes, and the phospholipid fatty acid method to determine microbial biomass and ratios of fungi:bacteria and Gram-positive:Gram-negative bacteria. RH did not respond to climate treatments but was greater in the warm and dry summer than in the cool and less-dry fall. These dynamics were better explained when microbial data were taken into account compared to when only physical data (temperature and moisture) were used. Overall, our results suggest that RH responses to temperature are stronger when soil contains more active microbes, and that seasonal patterns of RH can be better explained by shifts in microbial activity than by shifts in the relative abundances of fungi and Gram-positive and Gram-negative bacteria. These findings contribute to our understanding of how and under which conditions microbes influence soil C responses to climate.

Microbial dormancy in the marine subsurface: Global endospore abundance and response to burial.

Marine sediments host an unexpectedly large microbial biosphere, suggesting unique microbial mechanisms for surviving burial and slow metabolic turnover. Although dormancy is generally considered an important survival strategy, its specific role in subsurface sediments remains unclear. We quantified dormant bacterial endospores in 331 marine sediment samples from diverse depositional types and geographical origins. The abundance of endospores relative to vegetative cells increased with burial depth and endospores became dominant below 25 m, with an estimated population of 2.5 × 1028 to 1.9 × 1029 endospores in the uppermost kilometer of sediment and a corresponding biomass carbon of 4.6 to 35 Pg surpassing that of vegetative cells. Our data further identify distinct endospore subgroups with divergent resistance to burial and aging. Endospores may shape the deep biosphere by providing a core population for colonization of new habitats and/or through low-frequency germination to sustain slow growth in this environment.

Survival of the fewest: Microbial dormancy and maintenance in marine sediments through deep time.

Microorganisms buried in marine sediments are known to endure starvation over geologic timescales. However, the mechanisms of how these microorganisms cope with prolonged energy limitation is unknown and therefore yet to be captured in a quantitative framework. Here, we present a novel mathematical model that considers (a) the physiological transitions between the active and dormant states of microorganisms, (b) the varying requirement for maintenance power between these phases, and (c) flexibility in the provenance (i.e., source) of energy from exogenous and endogenous catabolism. The model is applied to sediments underlying the oligotrophic South Pacific Gyre where microorganisms endure ultra‐low fluxes of energy for tens of millions of years. Good fits between model simulations and measurements of cellular carbon and organic carbon concentrations are obtained and are interpreted as follows: (a) the unfavourable microbial habitat in South Pacific Gyre sediments triggers rapid mortality and a transition to dormancy; (b) there is minimal biomass growth, and organic carbon consumption is dominated by catabolism to support maintenance activities rather than new biomass synthesis; (c) the amount of organic carbon that microorganisms consume for maintenance activities is equivalent to approximately 2% of their carbon biomass per year; and (d) microorganisms must rely solely on exogenous rather than endogenous catabolism to persist in South Pacific Gyre sediments over long timescales. This leads us to the conclusion that under oligotrophic conditions, the fitness of an organism is determined by its ability to simply stay alive, rather than to grow. This modelling framework is designed to be flexible for application to other sites and habitats, and thus serves as a new quantitative tool for determining the habitability of and an ultimate limit for life in any environment.

The emergence of metabolic heterogeneity and diverse growth responses in isogenic bacterial cells.

Microorganisms adapt to frequent environmental changes through population diversification. Previous studies demonstrated phenotypic diversity in a clonal population and its important effects on microbial ecology. However, the dynamic changes of phenotypic composition have rarely been characterized. Also, cellular variations and environmental factors responsible for phenotypic diversity remain poorly understood. Here, we studied phenotypic diversity driven by metabolic heterogeneity. We characterized metabolic activities and growth kinetics of starved Escherichia coli cells subject to nutrient upshift at single-cell resolution. We observed three subpopulations with distinct metabolic activities and growth phenotypes. One subpopulation was metabolically active and immediately grew upon nutrient upshift. One subpopulation was metabolically inactive and non-viable. The other subpopulation was metabolically partially active, and did not grow upon nutrient upshift. The ratio of these subpopulations changed dynamically during starvation. A long-term observation of cells with partial metabolic activities indicated that their metabolism was later spontaneously restored, leading to growth recovery. Further investigations showed that oxidative stress can induce the emergence of a subpopulation with partial metabolic activities. Our findings reveal the emergence of metabolic heterogeneity and associated dynamic changes in phenotypic composition. In addition, the results shed new light on microbial dormancy, which has important implications in microbial ecology and biomedicine.

Evolution with a seed bank: The population genetic consequences of microbial dormancy.

Dormancy is a bet‐hedging strategy that allows organisms to persist through conditions that are suboptimal for growth and reproduction by entering a reversible state of reduced metabolic activity. Dormancy allows a population to maintain a reservoir of genetic and phenotypic diversity (i.e., a seed bank) that can contribute to the long‐term survival of a population. This strategy can be potentially adaptive and has long been of interest to ecologists and evolutionary biologists. However, comparatively little is known about how dormancy influences the fundamental evolutionary forces of genetic drift, mutation, selection, recombination, and gene flow. Here, we investigate how seed banks affect the processes underpinning evolution by reviewing existing theory, implementing novel simulations, and determining how and when dormancy can influence evolution as a population genetic process. We extend our analysis to examine how seed banks can alter macroevolutionary processes, including rates of speciation and extinction. Through the lens of population genetic theory, we can understand the extent that seed banks influence the evolutionary dynamics of microorganisms as well as other taxa.

Microbial dormancy promotes microbial biomass and respiration across pulses of drying-wetting stress

Recent work suggests that metabolic activation and deactivation of microbes in soil strongly influences soil carbon (C) dynamics and climate feedbacks. However, few soil C models consider these transitions. We hypothesized that microbes’ capacity to enter and exit dormancy in response to unfavorable and favorable environmental conditions decreases the sensitivity of microbial biomass and cumulative respiration to environmental stress. To test this hypothesis, we collected data from a rewetting experiment and used it to design and parameterize dormancy in an existing microbe-based soil C model. Then we compared predictions of microbial biomass and soil heterotrophic respiration (RH) under simulated cycles of stressful (dryness) and favorable (wet pulses) conditions. Because the influence of moisture on microbial processes in soil generally depends on temperature, we collected data and tested predictions at different temperatures. When dormancy was not taken into account, simulated microbial biomass and cumulative microbial respiration over five years were lower and decreased faster under lengthening drying-wetting cycles. Differences due to dormancy increased with temperature and with the length of the dry periods between wetting events. We conclude that ignoring both the capacity of microbes to enter and exit dormancy in response to the environment and the consequences of these metabolic responses for soil C cycling results in predictions of unrealistically low RH under warming and drying-wetting cycles.

Microscale Insight into Microbial Seed Banks.

Microbial dormancy leads to the emergence of seed banks in environmental, engineered, and host-associated ecosystems. These seed banks act as reservoirs of diversity that allow microbes to persist under adverse conditions, including extreme limitation of resources. While microbial seed banks may be influenced by macroscale factors, such as the supply of resources, the importance of microscale encounters between organisms and resource particles is often overlooked. We hypothesized that dimensions of spatial, trophic, and resource complexity determine rates of encounter, which in turn, drive the abundance, productivity, and size of seed banks. We tested this using >10,000 stochastic individual based models (IBMs) that simulated energetic, physiological, and ecological processes across combinations of resource, spatial, and trophic complexity. These IBMs allowed realistic dynamics and the emergence of seed banks from ecological selection on random variation in species traits. Macroscale factors like the supply and concentration of resources had little effect on resource encounter rates. In contrast, encounter rates were strongly influenced by interactions between dispersal mode and spatial structure, and also by the recalcitrance of resources. In turn, encounter rates drove abundance, productivity, and seed bank dynamics. Time series revealed that energetically costly traits can lead to large seed banks and that recalcitrant resources can lead to greater stability through the formation of seed banks and the slow consumption of resources. Our findings suggest that microbial seed banks emerge from microscale dimensions of ecological complexity and their influence on resource limitation and energetic costs.