title: High instability of a nematicidal Cry toxin plasmid in Bacillus thuringiensis.
abstract: In bacterial pathogens, virulence factors are often carried on plasmids and other mobile genetic elements, and as such, plasmid evolution is central in understanding pathogenicity. Bacillus thuringiensis is an invertebrate pathogen that uses plasmid-encoded crystal (Cry) toxins to establish infections inside the host. Our study aimed to quantify stability of two Cry toxin-encoding plasmids, BTI_23p and BTI_16p, under standard laboratory culturing conditions. These two plasmids are part of the genome of the B. thuringiensis strain MYBT18679, which is of particular interest because of its high pathogenicity towards nematodes. One of the plasmids, BTI_23p, was found to be highly unstable, with substantial loss occurring within a single growth cycle. Nevertheless, longer term experimental evolution in the absence of a host revealed maintenance of the plasmid at low levels in the bacterial populations. BTI_23p encodes two nematicidal Cry toxins, Cry21Aa2 and Cry14Aa1. Consistent with previous findings, loss of the plasmid abolished pathogenicity towards the nematode Caenorhabditis elegans, which could be rescued by addition of Cry21Aa2-expressing Escherichia coli. These results implicate BTI_23p as a plasmid that is required for successful infection, yet unstable when present at high frequency in the population, consistent with the role of Cry toxins as public goods.
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany. Electronic address: anna.sheppard@ndm.ox.ac.uk.
authors.forename: Anna E
authors.initials: AE
authors.lastname: Sheppard
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: Rania
authors.initials: R
authors.lastname: Nakad
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: Manja
authors.initials: M
authors.lastname: Saebelfeld
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: Anna C
authors.initials: AC
authors.lastname: Masche
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Dierking
authors.affiliation: Department of Evolutionary Ecology and Genetics, Christian-Albrechts University of Kiel, Kiel, Germany.
authors.forename: Hinrich
authors.initials: H
authors.lastname: Schulenburg
mesh_terms: D000818:Animals; D000969:Antinematodal Agents / Q000737:chemistry / Q000494:pharmacology; D001413:Bacillus thuringiensis / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000382:microbiology*; D004731:Endotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D006460:Hemolysin Proteins / Q000235:genetics*; D054884:Host-Pathogen Interactions; D010957:Plasmids / Q000235:genetics* / Q000502:physiology; D014774:Virulence / Q000235:genetics
chemical_list: D000969:Antinematodal Agents; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: Bacillus thuringiensis; Caenorhabditis elegans; Crystal toxin; Experimental evolution; Pathogenicity; Plasmid instability