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Xenorhabdus bovienii T228 phase variation and virulence are independent of RecA function

Microbiology and Immunology, Department of Molecular BioSciences, Adelaide University, Adelaide, South Australia 5005¹

Author for correspondence: Connor J. Thomas. Tel: +61 8 8303 5396. Fax: +61 8 8303 4362. e-mail: connor.thomas{at}adelaide.edu.au

Colony pleomorphism, or phase variation, expressed by entomopathogenic bacteria belonging to the genus Xenorhabdus, is an important factor which determines the association of the bacteria with their nematode symbiont and the outcome of infection of susceptible insect larvae by the bacterium– nematode parasitic complex. The mechanism underlying phase variation is unknown. To determine whether RecA-mediated processes are linked to phase variation, the recA gene of Xenorhabdus bovienii was cloned and sequenced. When expressed in a recA-deleted strain of Escherichia coli, the X. bovienii recA clone was able to complement the loss of RecA function. X. bovienii chromosomal recA insertion mutants showed increased sensitivity to UV. Phase 1 forms did not show altered ability to convert to phase 2 and no significant differences in expression of other phase-dependent characteristics, including phospholipase C, haemolysin, protease, antibiotic activity and Congo Red binding, were noted. Furthermore, the LD₅₀ of the X. bovienii recA insertion mutant for Galleria mellonella larvae was not significantly different from that of wild-type strains. From these data the authors conclude that recA is unlikely to be involved in phase variation, the expression of phase-dependent characteristics, or virulence factors involved in killing of susceptible larvae.

Keywords: entomopathogen, recombination, recA mutation, Galleria, phylogeny

Abbreviations: Ap, ampicillin; Km, kanamycin; NA, nutrient agar; NB, nutrient broth; P1, phase 1; P2, phase 2; Sm, streptomycin; Tc, tetracycline

The GenBank accession number for the sequence reported in this paper is U87924.

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