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Ecological drivers of intragenomic conflict resolution

Lead Research Organisation: University of Sheffield
Department Name: Animal and Plant Sciences

Abstract

The classical view of evolution is as a process of gradual accumulation of small changes by mutations passed on to descendants. However, genome sequencing has revealed that in bacteria genes are frequently exchanged between cells and species by a process called horizontal gene transfer, allowing evolution to make big, fast jumps. Genes often pass from one bacterial cell to another on circular pieces of DNA called plasmids. However, acquiring a plasmid can be costly to a bacterial cell because plasmids use energy and introduce new genes that disrupt the normal working of the bacterium. In evolutionary theory these costs are called intragenomic conflicts because what is good for the plasmid is bad for the bacterial chromosome, and vice versa. We have recently shown that evolution can resolve these conflicts by a process of 'compensatory evolution', whereby either the bacterial chromosome or the plasmid gains mutations that lessen the cost of carrying the plasmid. By increasing plasmid survival, compensatory evolution is likely to increase the chance of new genes jumping onto plasmids from bacterial chromosomes, allowing these to be shared with other species in the community. In this proposal we will discover how the ecology of microbial communities and the environments they live in shape the processes of compensatory evolution, and whether compensatory evolution itself speeds up the sharing of genes between species.

Planned Impact

Who will benefit from this research and how?

This is basic blue-skies research that will advance fundamental understanding of evolutionary processes and dynamics in bacterial communities. Nevertheless, bacterial evolution has a broad range of important impacts upon society, for example through the effects of rapid evolutionary change on the prognosis of clinical infections, the evolutionary emergence of antibiotic resistance, and evolutionary responses of microbial communities underpinning the functioning of ecosystems to environmental change. Despite the widespread and fundamental impact of rapid microbial evolution in general and horizontal gene transfer (HGT) in particular upon society, these evolutionary processes remain very poorly understood by the general public and policy-makers. The key benefits deriving from this research will therefore be increased knowledge and understanding of bacterial evolution among the following groups:

Secondary school age children: Teaching of evolution in Key Stages 2 and 3 of the National Curriculum is mainly theoretical and lacking in engaging practical classes. We will take experimental evolution into the school classroom allowing pupils to experience evolution in action themselves in real time, generating excitement about microbes and evolution and offering deeper experiential learning.

General public: Bacterial evolution is high on the news agenda due to the crisis in antimicrobial resistance (AMR), however few non-scientists realise that this societal problem is exacerbated by HGT-mediated evolution. Public engagement activities will enhance public understanding of HGT and put this into the context of AMR to show what we can all do to reduce the risks of AMR.

Policy makers in healthcare and agri-food sectors: HGT impacts the evolutionary emergence of AMR in the clinic and the spread of functional traits in soil bacterial communities. Designing policies and interventions that aim to e.g. limit the spread of AMR or conserve the functional diversity of soil bacterial communities, requires sharing knowledge and understanding of the dynamics of HGT and how these are shaped by the ecology of microbial communities and their environments arising from this research with stakeholders and policymakers in these sectors. We will engage with healthcare stakeholders via an established clinical network (PARC; PI Brockhurst is a member) and agri-food stakeholders via the N8 AgriFood Partnership facilitated by the N8 AgriFood Knowledge Exchange Fellows.

Publications

10 25 50

Related Projects

Project Reference Relationship Related To Start End Award Value
NE/R008825/1 01/02/2018 30/05/2020 £487,227
NE/R008825/2 Transfer NE/R008825/1 31/05/2020 30/10/2021 £149,022
 
Description Bacteria rapidly adapt to new environments increasing their fitness. We show that this process of environmental adaptation is prevented by the presence of other bacterial species in simple communities in soil microcosms. Published in Evolution Letters.
Acquisition of plasmids causes costs for the host cell which can be ameliorated by mutations elsewhere in the genome (compensatory mutations). Using mathematical models we show that plasmid survival is enhanced when these compensatory mutations are encoded on the plasmid itself, rather than on the bacterial chromosome. Published in mSystems. Using RNA sequencing to compare gene expression in bacteria with or without plasmids we show that the costs of plasmid acquisition are caused by genetic conflicts between genes on the chromosome and plasmid, causing deleterious expression of other mobile genetic elements in the bacterial chromosome. These effects are fixed by compensatory mutations, restoring normal gene expression. Published in PLOS Biology. Bacteria can rapidly adapt to the costs of acquiring plasmids by gaining compensatory mutations to overcome intragenomic conflict. Published in Microbiology. Moreover, a single chromosomal compensatory mutation can ameliorate the cost of carrying multiple plasmids in the same cell. Published in mBio.
The dynamics of plasmids in bacterial populations and communities are shaped by spatio-temporal variation selection for the traits carried by plasmids and also by the structure of the microbial community and the abilities of the constituent species to act as hosts for the plasmid(s). Published in Proceedings of the Royal Society B, FEMS Microbiology Ecology, and Microbiology.
We also published a major review on the ecology and evolution of pangenomes focusing on the importance of intragenomic conflicts and horizontal gene transfer. Published in Current Biology.
Exploitation Route These findings advance understanding of how species evolve in complex communities, which is relevant to understanding how species will adapt to anthropogenic environmental change. These findings advance understanding of how plasmids survive and spread in bacterial populations/communities, which is relevant to understanding the spread of antibiotic resistance genes. These findings reveal how bacterial genomes evolve through the horizontal acquisition of new genetic material from other cells and species.
Sectors Environment

Healthcare

 
Description The findings from this project have been delivered to the public through outreach activities and helped increase understanding of the role for microbes in health (of soils, plants and animals). The findings have fed into a Parliamentary Office of Science and Technology POSTnote. The finding have formed the basis for a public engagement comic book "Luna and Simon: Bizarre Bacteria and Peculiar Plasmids" about plasmids written by Edward Ross for the general public https://www.andthemicrobes.org
First Year Of Impact 2019
Sector Agriculture, Food and Drink,Creative Economy,Digital/Communication/Information Technologies (including Software),Environment,Culture, Heritage, Museums and Collections
Impact Types Cultural

Societal

Policy & public services

 
Title Analysis scripts for "Competitive species interactions constrain abiotic adaptation in a bacterial soil community" 
Description This record contains sample R analysis scripts associated with 'Competitive species interactions constrain abiotic adaptation in a bacterial soil community' by J. P. J. Hall et al. (2018). The data used in these analyses is presented in the Supplementary Tables published as an Excel workbook in the manuscript. For ease of analysis, .csv versions of these tables are provided in another ORDA record (doi: 10.15131/shef.data.7088906). Full details of the experiments and their interpretation can be found in the open access paper, published in Evolution Letters (see References below). Briefly, Pseudomonas fluorescens SBW25 carrying the mercury resistance plasmid pQBR57 were cultured in soil microcosms in the presence or absence of a competitor, Pseudomonas putida KT2440, and with or without mercury. After ~440 generations, clones were re-isolated and their fitness relative to the ancestral strain was measured. Clones from single-species treatments had increased relative fitness compared with those from co-culture. Investigation of the resequenced genomes showed parallel mutations occurring to the actP gene, but only in clones which had evolved without P. putida. Subsequent experiments show that actP disruption is associated with increased growth, but only where P. putida is absent. 1_CompetitionsAnalysis.R: example code describing analyses presented in Figure 1, showing relative fitness of evolved clones. 2_MutationsAnalysis.R: example code describing analyses presented in Figure 2, showing parallel mutations in evolved clones. 4_GrowthAnalysis.R: example code describing analyses presented in Figure 4, showing growth of clones with mutated and non-mutated actP in the presence and absence of P. putida. S3_PopulationsAnalysis.R: example code describing analyses presented in Figure S3, showing population densities of P. fluorescens during evolution experiment. S4_AcetateGrowth.R: example code describing analyses presented in Figure S4, showing growth of P. fluorescens in minimal media with glycerol or acetate as carbon sources. S5A_DiscDiffusion.R: example code describing analyses presented in Figure S5A, showing inhibition of evolved P. fluorescens by tellurite. S5B_TellMIC.R: example code describing analyses presented in Figure S5B, showing inhibition of evolved P. fluorescens by tellurite. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
URL https://figshare.shef.ac.uk/articles/dataset/Analysis_scripts_for_Competitive_species_interactions_c...
 
Title Analysis scripts for "Competitive species interactions constrain abiotic adaptation in a bacterial soil community" 
Description This record contains sample R analysis scripts associated with 'Competitive species interactions constrain abiotic adaptation in a bacterial soil community' by J. P. J. Hall et al. (2018). The data used in these analyses is presented in the Supplementary Tables published as an Excel workbook in the manuscript. For ease of analysis, .csv versions of these tables are provided in another ORDA record (doi: 10.15131/shef.data.7088906). Full details of the experiments and their interpretation can be found in the open access paper, published in Evolution Letters (see References below). Briefly, Pseudomonas fluorescens SBW25 carrying the mercury resistance plasmid pQBR57 were cultured in soil microcosms in the presence or absence of a competitor, Pseudomonas putida KT2440, and with or without mercury. After ~440 generations, clones were re-isolated and their fitness relative to the ancestral strain was measured. Clones from single-species treatments had increased relative fitness compared with those from co-culture. Investigation of the resequenced genomes showed parallel mutations occurring to the actP gene, but only in clones which had evolved without P. putida. Subsequent experiments show that actP disruption is associated with increased growth, but only where P. putida is absent. 1_CompetitionsAnalysis.R: example code describing analyses presented in Figure 1, showing relative fitness of evolved clones. 2_MutationsAnalysis.R: example code describing analyses presented in Figure 2, showing parallel mutations in evolved clones. 4_GrowthAnalysis.R: example code describing analyses presented in Figure 4, showing growth of clones with mutated and non-mutated actP in the presence and absence of P. putida. S3_PopulationsAnalysis.R: example code describing analyses presented in Figure S3, showing population densities of P. fluorescens during evolution experiment. S4_AcetateGrowth.R: example code describing analyses presented in Figure S4, showing growth of P. fluorescens in minimal media with glycerol or acetate as carbon sources. S5A_DiscDiffusion.R: example code describing analyses presented in Figure S5A, showing inhibition of evolved P. fluorescens by tellurite. S5B_TellMIC.R: example code describing analyses presented in Figure S5B, showing inhibition of evolved P. fluorescens by tellurite. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
URL https://figshare.shef.ac.uk/articles/dataset/Analysis_scripts_for_Competitive_species_interactions_c...
 
Title Data for "Competitive species interactions constrain abiotic adaptation in a bacterial soil community" 
Description This record contains .csv-formatted Supplementary Data tables associated with 'Competitive species interactions constrain abiotic adaptation in a bacterial soil community' by J. P. J. Hall et al. (2018). Sample analysis scripts for this data are provided in another ORDA record, doi:10.15131/shef.data.7043279. SuppTab3.csv: .csv version of Supplementary Table 3. Fitness of evolved pQBR57-bearing Pseudomonas fluorescens SBW25 clones relative to their ancestor in soil microcosms. Six or seven clones were tested from populations evolved with or without mercury, and with or without Pseudomonas putida KT2440 competitors. Fitness was measured both with and without mercury. evo_rep = evolutionary replicate; cln_rep = clone reference; mrk = marker; cul = single-species (ss) or co-culture (co); evo_mer = mercury treatment in evolution experiment; test_mer = mercury in test conditions; fitness = relative fitness compared with ancestor (per day); fitness (generation) = relative fitness compared with ancestor (per generation). SuppTab4.csv: .csv version of Supplementary Table 4. Mutations in actP detected in evolved Pseudomonas fluorescens SBW25 clones. sample_number = sample number in sequencing run; cln_name = code referring to that clone; evo_rep = evolutionary replicate; cln_rep = clone reference; pla = P if clone has pQBR57; evo_mer = mercury treatment in evolution experiment; pla_trt = plasmid treatment; mrk = marker; cul = cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); mutation_position = bp affected by mutation; effect_position = codon affected by position SuppTab5.csv: .csv version of Supplementary Table 5. Growth of sequenced evolved Pseudomonas fluorescens SBW25 clones in soil microcosms with and without Pseudomonas putida KT2440 competitors. sample_number = sample number in sequencing run; cln_name = code referring to that clone; evo_rep = evolutionary replicate; cln_rep = clone reference; evo_mer = mercury treatment in evolution experiment; pla_trt = plasmid treatment; mrk = marker; cul = cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); mutation_position = bp affected by mutation; effect_position = codon affected by position; tech_rep = replicate experiment in which this clone was tested; m.with_putida = growth rate in co-culture (per day); m.alone = growth rate in single-species culture (per day); w = m.with_putida/m.alone SuppTab6.csv: .csv version of Supplementary Table 6. Population density at transfer of Pseudomonas fluorescens SBW25 from the evolution experiment. rep = evolutionary replicate; tra = transfer; spc = species; cul = single-species (ss) or co-culture (co); pla_trt = plasmid treatment; mer = mercury treatment; mrk = marker; tot = cfu/g at transfer SuppTab7.csv: .csv version of Supplementary Table 7. Relative growth of evolved Pseudomonas fluorescens SBW25 clones in acetate minimal media relative to glycerol minimal media. sample_number = sample number in sequencing run; cln_name = code referring to that clone; actp.status = disrupted or wild-type; actp.mutation = impact of the mutation on actP; technical.rep = replicate experiment in which this clone was tested; ratio = m_acetate/m_glycerol; difference = m_acetate - m_glycerol. SuppTab8.csv: .csv version of Supplementary Table 8. Zones of inhibition produced by tellurite-impregnated discs for evolved Pseudomonas fluorescens SBW25 clones. sample_number = sample number in sequencing run; cln_name = code referring to that clone; tech_rep = replicate experiment in which this clone was tested; evo_mer = mercury treatment in evolution experiment; plasmid_trt = plasmid treatment; cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); actp_status = disrupted or wild-type; area = area of zone of inhibition (mm^2) SuppTab9.csv: .csv version of Supplementary Table 9. Growth of evolved Pseudomonas fluorescens SBW25 clones in various concentrations of tellurite. sample_number = sample number in sequencing run; cln_name = code referring to that clone; tech_rep = replicate experiment in which this clone was tested; tell_conc = tellurite concentration (µg/ml); OD600_scaled = OD600 scaled between 0 and 1 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
URL https://figshare.shef.ac.uk/articles/dataset/Data_for_Competitive_species_interactions_constrain_abi...
 
Title Data for "Competitive species interactions constrain abiotic adaptation in a bacterial soil community" 
Description This record contains .csv-formatted Supplementary Data tables associated with 'Competitive species interactions constrain abiotic adaptation in a bacterial soil community' by J. P. J. Hall et al. (2018). Sample analysis scripts for this data are provided in another ORDA record, doi:10.15131/shef.data.7043279. SuppTab3.csv: .csv version of Supplementary Table 3. Fitness of evolved pQBR57-bearing Pseudomonas fluorescens SBW25 clones relative to their ancestor in soil microcosms. Six or seven clones were tested from populations evolved with or without mercury, and with or without Pseudomonas putida KT2440 competitors. Fitness was measured both with and without mercury. evo_rep = evolutionary replicate; cln_rep = clone reference; mrk = marker; cul = single-species (ss) or co-culture (co); evo_mer = mercury treatment in evolution experiment; test_mer = mercury in test conditions; fitness = relative fitness compared with ancestor (per day); fitness (generation) = relative fitness compared with ancestor (per generation). SuppTab4.csv: .csv version of Supplementary Table 4. Mutations in actP detected in evolved Pseudomonas fluorescens SBW25 clones. sample_number = sample number in sequencing run; cln_name = code referring to that clone; evo_rep = evolutionary replicate; cln_rep = clone reference; pla = P if clone has pQBR57; evo_mer = mercury treatment in evolution experiment; pla_trt = plasmid treatment; mrk = marker; cul = cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); mutation_position = bp affected by mutation; effect_position = codon affected by position SuppTab5.csv: .csv version of Supplementary Table 5. Growth of sequenced evolved Pseudomonas fluorescens SBW25 clones in soil microcosms with and without Pseudomonas putida KT2440 competitors. sample_number = sample number in sequencing run; cln_name = code referring to that clone; evo_rep = evolutionary replicate; cln_rep = clone reference; evo_mer = mercury treatment in evolution experiment; pla_trt = plasmid treatment; mrk = marker; cul = cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); mutation_position = bp affected by mutation; effect_position = codon affected by position; tech_rep = replicate experiment in which this clone was tested; m.with_putida = growth rate in co-culture (per day); m.alone = growth rate in single-species culture (per day); w = m.with_putida/m.alone SuppTab6.csv: .csv version of Supplementary Table 6. Population density at transfer of Pseudomonas fluorescens SBW25 from the evolution experiment. rep = evolutionary replicate; tra = transfer; spc = species; cul = single-species (ss) or co-culture (co); pla_trt = plasmid treatment; mer = mercury treatment; mrk = marker; tot = cfu/g at transfer SuppTab7.csv: .csv version of Supplementary Table 7. Relative growth of evolved Pseudomonas fluorescens SBW25 clones in acetate minimal media relative to glycerol minimal media. sample_number = sample number in sequencing run; cln_name = code referring to that clone; actp.status = disrupted or wild-type; actp.mutation = impact of the mutation on actP; technical.rep = replicate experiment in which this clone was tested; ratio = m_acetate/m_glycerol; difference = m_acetate - m_glycerol. SuppTab8.csv: .csv version of Supplementary Table 8. Zones of inhibition produced by tellurite-impregnated discs for evolved Pseudomonas fluorescens SBW25 clones. sample_number = sample number in sequencing run; cln_name = code referring to that clone; tech_rep = replicate experiment in which this clone was tested; evo_mer = mercury treatment in evolution experiment; plasmid_trt = plasmid treatment; cul = single-species (ss) or co-culture (co); mutation = actP mutation; effect = effect of mutation; impact = impact of mutation (defined by SNPeff); actp_status = disrupted or wild-type; area = area of zone of inhibition (mm^2) SuppTab9.csv: .csv version of Supplementary Table 9. Growth of evolved Pseudomonas fluorescens SBW25 clones in various concentrations of tellurite. sample_number = sample number in sequencing run; cln_name = code referring to that clone; tech_rep = replicate experiment in which this clone was tested; tell_conc = tellurite concentration (µg/ml); OD600_scaled = OD600 scaled between 0 and 1 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
URL https://figshare.shef.ac.uk/articles/dataset/Data_for_Competitive_species_interactions_constrain_abi...
 
Description British Science Week 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact The andthemicrobes.org microbiology comics were featured in an interactive online learning workshop resource called Biodiscovery that was part of British Science week. The site received 100s of visitors.
Year(s) Of Engagement Activity 2022
URL http://www.bit.ly/biodiscovery
 
Description Community Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Our andthemicrobes.org "Lune and Simon" microbiology comics were used at a major science outreach festival at Manchester Library organised by the University of Manchester.
Year(s) Of Engagement Activity 2022
URL https://www.mub.eps.manchester.ac.uk/sciencex/
 
Description Interactive microbiome exhibit (Great Yorkshire Show) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 630 people visited our interactive exhibit about the important role that microbiomes play in health (in general encompassing soils, plants and animals) as part of the Discovery Zone. This was aimed at secondary school children but attended by a wider range of ages including adults. The exhibit sparked questions and discussion.
Year(s) Of Engagement Activity 2019
 
Description Public engagement comic book 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Comic book "Luna and Simon: Bizarre Bacteria and Peculiar Plasmids" written by Edward Ross based on our research and accompanying website. Printed and online. Production featured workshops for 20 school children. Over 500 printed copies have been distributed and far more downloaded from the website.
Year(s) Of Engagement Activity 2020
URL https://www.andthemicrobes.org
 
Description School Visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Talk and discussion with ~50 pupils at Bramhall High School in Stockport about phages and phage therapy using the andthemicrobes.org comics as lesson tools
Year(s) Of Engagement Activity 2023
URL https://www.andthemicrobes.org
 
Description Schools using outreach resources for lessons 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Our microbiology outreach comics were used as learning resources in both art and science lessons at Plymouth Grove Primary School
Year(s) Of Engagement Activity 2022
URL https://www.andthemicrobes.org
 
Description Sustaining the soil microbiome POSTnote 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Parliamentary Office of Science and Technology Note (POSTnote) entitled "Susutaining the soil microbiome" authored by Cagla Stevenson (PhD student) in my group
Year(s) Of Engagement Activity 2019
URL https://researchbriefings.parliament.uk/ResearchBriefing/Summary/POST-PN-0601
 
Description Workshops for a public engagement comic book 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Workshop for a group of school age children to develop a comic book about microbes and their mobile genetic elements. Involved creative tasks and discussion around scientific topics depicted in the draft comic book. Sparked enthusiastic discussion and questions.

Comic book to be published in 2021.
Year(s) Of Engagement Activity 2019,2020