Summary: Expression of Bacillus thuringiensis delta‐endotoxins has proven to be a successful strategy for obtaining insect resistance in transgenic plants. Drawbacks of expression of a single resistance gene are the limited target spectrum and the potential for rapid adaptation of the pest. Hybrid toxins with a wider target spectrum in combination with existing toxins may be used as tool to mitigate these problems. In this study, Desiree potato plants were genetically modified to resist attack by insect species belonging to the orders Coleoptera and Lepidoptera, through the insertion of such a hybrid gene, SN19. Transgenic plants were shown to be resistant against Colorado potato beetle larvae and adults, potato tuber moth larvae, and European corn borer larvae. These are the first transgenic plants resistant to pests belonging to two different insect orders. In addition, the target receptor recognition of this hybrid protein is expected to be different from Cry proteins currently in use for these pests. This makes it a useful tool for resistance management strategies.

Cry15Aa protein, produced byBacillus thuringiensisserovar thompsoni HD542 in a crystal together with a 40-kDa accompanying protein, is one of a small group of nontypical, less well-studied members of the Cry family of insecticidal proteins and may provide an alternative for the more commonly used Cry proteins in insect pest management. In this paper, we describe the characterization of the Cry15Aa and 40-kDa protein's biochemical and insecticidal properties and the mode of action. Both proteins were solubilized above pH 10 in vitro. Incubation of solubilized crystal proteins with trypsin or insect midgut extracts rapidly processed the 40-kDa protein to fragments too small to be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whereas the Cry15 protein yielded a stable product of approximately 30 kDa. Protein N-terminal sequencing showed that Cry15 processing occurs exclusively at the C-terminal end. Cry15 protein showed in vitro hemolytic activity, which was greatly enhanced by preincubation with trypsin or insect gut extract. Larvae of the lepidopteran insectsManduca sexta,Cydia pomonella, andPieris rapaewere susceptible to crystals, and presolubilization of the crystals enhanced activity toP. rapae. Activity for all three species was enhanced by preincubation with trypsin. Larvae ofHelicoverpa armigeraandSpodoptera exiguawere relatively insensitive to crystals, and activity against these insects was not enhanced by prior solubilization or trypsin treatment. The 40-kDa crystal protein showed no activity in the insects tested, nor did its addition or coexpression inEscherichia coliincrease the activity of Cry15 in insecticidal and hemolytic assays.