Biology subjects

Rodriguez-Verdugo, A.

Publications and source records attributed to Rodriguez-Verdugo, A..

7 recordsLinked to original sources

bioRxiv2026

Microbial cross-feeding interactions reshape evolutionary trajectories of consumers by preserving motility.

Cross-feeding interactions, in which a producer species release by-products that serve as resources for a consumer species, play an important role in shaping microbial community diversity. Producers create opportunities for consumers by supplying high-energy resources that are often scarce in the environment. However, they also exert strong top-down effects by releasing metabolites in pulses and generating spatial gradients of resource availability. How these spatiotemporal constraints shape consumer evolution remains poorly understood. To address this question, we used a two-species cross-feeding system in which Acinetobacter johnsonii excretes benzoate (a by-product of benzyl alcohol oxidation) into the external environment where it is consumed by Pseudomonas putida. To assess how the origin of benzoate (externally supplied or produced by cross-feeding) shapes consumer evolution, we evolved P. putida for 200 generations in monoculture or in co-culture with A. johnsonii. Populations evolved in monoculture exhibited improved growth relative to the ancestor, whereas populations evolved under cross-feeding showed little to no growth improvement. Whole-genome sequencing revealed pervasive loss-of-function mutations in flagellar genes among populations evolved in monoculture, but not under cross-feeding conditions. High-throughput imaging assays showed that populations evolved under cross-feeding not only maintained but also enhanced functional motility. Competition experiments with single mutants revealed context-dependent fitness effects: loss-of-function mutations were highly beneficial when benzoate was externally supplied but deleterious when benzoate was supplied by A. johnsonii, highlighting the importance of motility in cross-feeding interactions. Together, our results show that resource origin fundamentally reshapes selective pressures and alters evolutionary outcomes in microbial communities. Significance StatementCross-feeding interactions are pervasive in microbial communities, often arising from metabolite leakage into the environment. Our study focused on understanding how cross-feeding interactions shape the evolution of populations that cross-feed on these resources. Using a highly trackable cross-feeding system and linking genotypic to phenotypic changes, we showed that cross-feeding interactions reshape the evolutionary trajectories of consumer species. Notably, selection acts on flagellar genes, but their effects on function depend on the origin of resources. When resources are externally supplied, selection favors loss of flagellar motility, whereas when resources are generated through cross-feeding, selection not only maintains functional motility but also enhance it. These findings highlight that navigating the environment is essential for exploiting high-energy resources generated through cross-feeding.

microbiology↗

bioRxiv2026

Unilateral cross-feeding constrains adaptive evolution, even in the producer without direct fitness effects.

Cross-feeding interactions are pervasive in microbial communities and profoundly shape community structure, stability, and function. While previous studies have explored how cross-feeding affects evolvability, this work has predominantly focused on bidirectional mutualistic interactions in engineered auxotrophic systems where both partners reciprocally exchange essential metabolites. However, most metabolic interactions in natural microbial communities are unidirectional, with organisms feeding on the metabolic waste products of other species. Our study addresses this gap by examining how a unidirectional cross-feeding interaction affects the evolutionary dynamics of both the producer (Acinetobacter johnsonii) and consumer (Pseudomonas putida) over 800 generations of experimental evolution. We found that co-culture constrained adaptive evolution in both species. Co-cultures exhibited lower {pi}N/{pi}S ratios (0.75 for P. putida; 1.04 for A. johnsonii) than monocultures (1.44 and 2.02, respectively) indicating stronger purifying selection against nonsynonymous mutations in the community context. Lineage tracking through whole genome sequencing of populations and clones revealed greater lineage diversity and complexity in monocultures, with more mutations showing significant parallelism across replicate populations. Additionally, P. putida evolved increased dependence on its partner; co-culture-evolved P. putida grew significantly worse than its ancestor when A. johnsonii was removed. These findings demonstrate that ecological interactions fundamentally reshape fitness landscapes and constrain adaptive evolution even when fitness benefits are unidirectional, with implications for understanding microbial community stability and predicting evolutionary dynamics in complex communities.

evolutionary biology↗

bioRxiv2024

Cross-feeding maintains diversification in an experimentally evolved microbial community

Microbial communities are incredibly diverse. Yet, the eco-evolutionary processes originating and maintaining this diversity remain understudied. Here, we investigate the patterns of diversification for Pseudomonas putida evolving in isolation and with Acinetobacter johnsonii leaking resources used by P. putida. We experimentally evolved four experimental replicates in monoculture and co-culture for 200 generations. We observed that P. putida diversified into two distinct morphotypes that differed from their ancestor by single-point mutations. One of the most prominent mutations hit the fleQ gene encoding the master regulator of flagella and biofilm formation. We experimentally confirmed that fleQ mutants were unable to swim and formed less biofilm than their ancestor, but they also produced higher yields. Interestingly, the fleQ genotype and other mutations swept to fixation in monocultures but not in co-cultures. In co-cultures, the two lineages stably coexisted for approximately 150 generations. We hypothesized that A. johnsonii modulates the coexistence of the two lineages through frequency-dependent selection. However, invasion experiments with two genotypes in monoculture and co-culture did not support this hypothesis. Instead, we found that, at the population level, the two morphotypes coexisted at similar relative abundances in the presence of A. johnsonii whereas, in its absence, one of the morphotypes was overrepresented in the population. Overall, our study suggests that interspecies interactions play an important role in shaping patterns of diversification in microbial communities. ImportanceIn nature, bacteria live in microbial communities and interact with other species, for example, through the exchange of resources leaked into the external environment (i.e., cross-feeding interactions). The role that these cross-feeding interactions play in shaping patterns of diversification remains understudied. Using a simple bacterial system in which one species cross-feeds resources to a second species (commensal species), we showed that the commensal species diversified into two subpopulations that persisted only when the cross-feeder partner was present. We further observed loss-of-function mutations in flagellar genes that were fixed in monocultures but not in co-cultures. Our findings suggest that cross-feeding species influence patterns of diversification of other species. Given that nutrient leakage is pervasive in microbial communities, the findings from this study have the potential to extend beyond our specific bacterial system. Importantly, our study has contributed to answering the larger question of whether species evolved differently in isolation versus when interacting with other species.

evolutionary biology↗

bioRxiv2024

Evolution Under Thermal Stress Affects Escherichia colis Resistance to Antibiotics

Exposure to both antibiotics and temperature changes can induce similar physiological responses in bacteria. Thus, changes in growth temperature may affect antibiotic resistance. Previous studies have found that evolution under antibiotic stress causes shifts in the optimal growth temperature of bacteria. However, little is known about how evolution under thermal stress affects antibiotic resistance. We examined 100+ heat-evolved strains of Escherichia coli that evolved under thermal stress. We asked whether evolution under thermal stress affects optimal growth temperature, if there are any correlations between evolving in high temperatures and antibiotic resistance, and if these strains antibiotic efficacy changes depending on the local environments temperature. We found that: (1) surprisingly, most of the heat-evolved strains displayed a decrease in optimal growth temperature and overall growth relative to the ancestor strain, (2) there were complex patterns of changes in antibiotic resistance when comparing the heat-evolved strains to the ancestor strain, and (3) there were few significant correlations among changes in antibiotic resistance, optimal growth temperature, and overall growth. ImportanceEscherichia coli, a bacteria species often found within the intestinal tract of warm-blooded organisms, can be harmful to humans. Like all species of bacteria, E. coli can evolve, particularly in the presence of stressful conditions such as extreme temperatures or antibiotic treatments. Recent evidence suggests that when encountering one source of stress, an organisms ability to deal with a different source of stress is also affected. With global climate change and the continued evolution of antibiotic-resistant bacteria, the need to further investigate how temperature and antibiotics interact is clear. The significance of our research is in identifying possible correlations between temperature and antibiotic stress, broadening our understanding of how stressors affect organisms, and allowing for insights into possible future evolutionary pathways.

evolutionary biology↗

bioRxiv2023

Substrate complexity buffers negative interactions in a synthetic microbial community of leaf litter degraders.

Microbes associated with leaf litter, the top layer of soil, collectively decompose organic matter such as plant polysaccharides, and respire carbon dioxide, regulating the land-atmosphere fluxes of carbon. Therefore, it is crucial to understand the processes limiting biopolymer degradation and their influences on soil community properties. For example, it is still unclear how substrate complexity - defined as the structure of the saccharide and the amount of external processing by extracellular enzymes - influences species interactions and species coexistence. Here, we tested the hypothesis that growth on monosaccharides (i.e., xylose) promotes negative interactions through resource competition, and growth on polysaccharides (i.e., xylan) promotes neutral or positive interactions through resource partitioning or synergism among extracellular enzymes. We assembled a three-species community of leaf litter-degrading bacteria isolated from a grassland site in Southern California. In the polysaccharide xylan, pairs of species stably coexisted and grew equally in co-culture and in monoculture. Conversely, in the monosaccharide xylose, competitive exclusion and negative interactions prevailed. These pairwise dynamics remained consistent in a three-species community: all three species coexisted in xylan, while only two species coexisted in xylose. A mathematical model parameterized from single-species growth behaviors showed that in xylose these dynamics could be explained by resource competition. Instead, the resource competition model could not predict the coexistence patterns in xylan. Overall, our study shows that substrate complexity influences species interactions and patterns of coexistence in a synthetic microbial community of leaf litter degraders that can serve as a model for studying carbon cycling and climate change. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/573222v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@abf5aeorg.highwire.dtl.DTLVardef@18e99d5org.highwire.dtl.DTLVardef@1158650org.highwire.dtl.DTLVardef@252623_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2020

Antibiotics shift the temperature response curve of Escherichia coli growth

Temperature variation--through time and across climatic gradients--affects individuals, populations, and communities. Yet how the thermal response of biological systems is altered by environmental stressors is poorly understood. Here we quantify two key features--optimal temperature and temperature breadth--to investigate how temperature responses vary in the presence of antibiotics. We use high-throughput screening to measure growth of Escherichia coli under single and pairwise combinations of 12 antibiotics across seven temperatures that range from 22{degrees}C to 46{degrees}C. We find that antibiotic stress often results in considerable changes in the optimal temperature for growth and a narrower temperature breadth. The direction of the optimal temperature shifts can be explained by the similarities between antibiotic-induced and temperature-induced damage to the physiology of the bacterium. We also find that the effects of pairs of stressors in the temperature response can often be explained by just one antibiotic out of the pair. Our study has implications for a general understanding of how ecological systems adapt and evolve to environmental changes.

ecology↗

bioRxiv2016

Adaptive mutations in RNA polymerase and the transcriptional terminator Rho have similar effects on Escherichia coli gene expression

Modifications to transcriptional regulators play a major role in adaptation. Here we compared the effects of multiple beneficial mutations within and between Escherichia coli rpoB, the gene encoding the RNA polymerase {beta} subunit, and rho, which encodes a transcriptional terminator. These two genes have harbored adaptive mutations in numerous E. coli evolution experiments but particularly in our previous large-scale thermal stress experiment, where the two genes characterized two alternative adaptive pathways. To compare the effects of beneficial mutations, we engineered four advantageous mutations into each of the two genes and measured their effects on fitness, growth, gene expression and transcriptional termination at 42.2{degrees}C. Among the eight mutations, two rho mutations had no detectable effect on relative fitness, suggesting they were beneficial only in the context of epistatic interactions. The remaining six mutations had an average relative fitness benefit of [~]20%. The rpoB mutations altered the expression of [~]1700 genes; rho mutations altered the expression of fewer genes, most of which were a subset of the genes altered by rpoB. Across our eight mutants, relative fitness correlated with the degree to which a mutation restored gene expression back to the unstressed, 37.0{degrees}C state. The rho mutations do not enhance transcriptional termination in known rho-terminated regions, but the genome-wide effects of mutations in both genes was to enhance termination. Although beneficial mutations in the two genes did not have identical effects on fitness, growth or gene expression, they acted predominantly through parallel phenotypic effects on gene expression and transcriptional termination.

evolutionary biology↗