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Applied and Environmental Microbiology, June 2003, p. 3344-3349, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3344-3349.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Interactions of Insecticidal Toxin Gene Products from Xenorhabdus nematophilus PMFI296

Martin Sergeant,1* Paul Jarrett,2 Margaret Ousley,1 and J. Alun W. Morgan1

Department of Plant Pathology and Microbiology,1 Entomological Sciences, Horticulture Research International, Wellesbourne, Warwick CV35 9EF, United Kingdom2

Received 13 December 2002/ Accepted 11 March 2003

Four genes on a genomic fragment from Xenorhabdus nematophilus PMFI296 were shown to be involved in insecticidal activity towards three commercially important insect species. Each gene was expressed individually and in combinations in Escherichia coli, and the insecticidal activity of the lysates was determined. The combined four genes (xptA1, xptA2, xptB1, and xptC1), in E. coli, showed activity towards Pieris brassicae, Pieris rapae, and Heliothis virescens. The genes xptA1, xptB1, and xptC1 were involved in expressing activity towards P. rapae and P. brassicae, while the genes xptA2, xptB1, and xptC1 were needed for activity towards H. virescens. When each of these three genes was expressed individually in E. coli and the cell lysates were used in insect assays or mixed and then used, insecticidal activity was detected at a very low level. If the genes xptB1 and xptC1 were expressed in the same E. coli cell and this cell lysate was mixed with cells expressing xptA1, activity was restored to P. rapae and P. brassicae. Similarly mixing XptB1/C1 lysate with XptA2 lysate restored activity towards H. virescens. Individual gene disruptions in X. nematophilus PMFI296 reduced activity to insects; this activity was restored by complementation with cells expressing either xptA1 or xptA2 for their respective disruptions or E. coli expressing both xptB1 and xptC1 for individual disruptions of either of these genes. The genes xptA2, xptC1, and xptB1 were expressed as an operon in PMFI296 and inactivation of xptA2 or xptC1 resulted in silencing of downstream gene(s), while xptA1 was expressed as a single gene. Therefore, the two three gene product combinations interact with each other to produce good insecticidal activity.

* Corresponding author. Mailing address: Department of Plant Pathology and Microbiology, Horticulture Research International, Wellesbourne, Warwick, United Kingdom. Phone: 44(0) 1789 470382. Fax: 44(0) 1789 470552. E-mail: martin.sergeant{at}hri.ac.uk.

Applied and Environmental Microbiology, June 2003, p. 3344-3349, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3344-3349.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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