Elucidation of the Photorhabdus temperata Genome and Generation of a Transposon Mutant Library To Identify Motility Mutants Altered in Pathogenesis
The entomopathogenic nematode Heterorhabditis bacteriophora forms a specific mutualistic association with its bacterial partner Photorhabdus temperata. The microbial symbiont is required for nematode growth and development, and symbiont recognition is strain specific. The aim of this study was to sequence the genome of P. temperata and identify genes that plays a role in the pathogenesis of the Photorhabdus-Heterorhabditis symbiosis. A draft genome sequence of P. temperata strain NC19 was generated. The 5.2-Mb genome was organized into 17 scaffolds and contained 4,808 coding sequences (CDS). A genetic approach was also pursued to identify mutants with altered motility. A bank of 10,000 P. temperata transposon mutants was generated and screened for altered motility patterns. Five classes of motility mutants were identified: (i) nonmotile mutants, (ii) mutants with defective or aberrant swimming motility, (iii) mutant swimmers that do not require NaCl or KCl, (iv) hyperswimmer mutants that swim at an accelerated rate, and (v) hyperswarmer mutants that are able to swarm on the surface of 1.25% agar. The transposon insertion sites for these mutants were identified and used to investigate other physiological properties, including insect pathogenesis. The motility-defective mutant P13-7 had an insertion in the RNase II gene and showed reduced virulence and production of extracellular factors. Genetic complementation of this mutant restored wild-type activity. These results demonstrate a role for RNA turnover in insect pathogenesis and other physiological functions.
Journal of Bacteriology
American Society for Microbiology
Digital Object Identifier (DOI)
Scientific Contribution Number
Sheldon Hurst IV, Holli Rowedder, Brandye Michaels, Hannah Bullock, Ryan Jackobeck, Feseha Abebe-Akele, Umjia Durakovic, Jon Gately, Erik Janicki and Louis S. Tisa. Elucidation of the Photorhabdus temperata Genome and Generation of a Transposon Mutant Library To Identify Motility Mutants Altered in Pathogenesis. J. Bacteriol. July 2015, vol. 197 no. 13 2201-2216. https://dx.doi.org/10.1128/JB.00197-15
Copyright © 2015, American Society for Microbiology. All Rights Reserved.
This is an article published by American Society for Microbiology in Journal of Bacteriology in 2015, available online: https://dx.doi.org/10.1128/JB.00197-15