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Journal of Bacteriology, September 2000, p. 5127-5138, Vol. 182, No. 18
Biocontrol and Biosecurity, Grasslands
Division, AgResearch, Lincoln,1 and
Department of Microbiology, University of Otago,
Dunedin,2 New Zealand
Received 3 March 2000/Accepted 26 June 2000
Serratia entomophila and Serratia
proteamaculans cause amber disease in the grass grub
Costelytra zealandica (Coleoptera: Scarabaeidae), an
important pasture pest in New Zealand. Larval disease symptoms include
cessation of feeding, clearance of the gut, amber coloration, and
eventual death. A 115-kb plasmid, pADAP, identified in S. entomophila is required for disease causation and, when
introduced into Escherichia coli, enables that organism to
cause amber disease. A 23-kb fragment of pADAP that conferred disease-causing ability on E. coli and a pADAP-cured strain
of S. entomophila was isolated. Using insertion
mutagenesis, the pathogenicity determinants were mapped to a 17-kb
region of the clone. Sequence analysis of the 17-kb region showed that
the predicted products of three of the open reading frames
(sepA, sepB, and sepC) showed
significant sequence similarity to components of the insecticidal toxin
produced by the bacterium Photorhabdus luminescens.
Transposon insertions in sepA, sepB, or
sepC completely abolished both gut clearance and cessation
of feeding on the 23-kb clone; when recombined back into pADAP, they
abolished gut clearance but not cessation of feeding. These results
suggest that SepA, SepB, and SepC together are sufficient for amber
disease causation by S. entomophila and that another locus
also able to exert a cessation-of-feeding effect is encoded elsewhere
on pADAP.
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Plasmid-Located Pathogenicity Determinants of Serratia
entomophila, the Causal Agent of Amber Disease of Grass Grub, Show
Similarity to the Insecticidal Toxins of Photorhabdus
luminescens
*
Corresponding author. Mailing address: AgResearch, P.O.
Box 60, Lincoln, New Zealand. Phone: 64 3 983 3985. Fax: 64 3 983 3946. E-mail: HurstM{at}Agresearch.cri.nz.
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