High instability of a nematicidal Cry toxin plasmid in Bacillus thuringiensis

Anna E. Sheppard, Rania Nakad, Manja Saebelfeld, Anna Cäcilia Masche, Katja Dierking, Hinrich Schulenburg

Research output: Contribution to journalJournal articleResearchpeer-review


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.
Original languageEnglish
JournalJournal of Invertebrate Pathology
Pages (from-to)34-40
Number of pages7
Publication statusPublished - 2016
Externally publishedYes


  • Bacillus thuringiensis
  • Crystal toxin
  • Pathogenicity
  • Plasmid instability
  • Experimental evolution
  • Caenorhabditis elegans


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