Expressed sequence tags from larval gut of the European corn borer (Ostrinia nubilalis): exploring candidate genes potentially involved in Bacillus thuringiensis toxicity and resistance.

BACKGROUND: Lepidoptera represents more than 160,000 insect species which include some of the most devastating pests of crops, forests, and stored products. However, the genomic information on lepidopteran insects is very limited. Only a few studies have focused on developing expressed sequence tag (EST) libraries from the guts of lepidopteran larvae. Knowledge of the genes that are expressed in the insect gut are crucial for understanding basic physiology of food digestion, their interactions with Bacillus thuringiensis (Bt) toxins, and for discovering new targets for novel toxins for use in pest management. This study analyzed the ESTs generated from the larval gut of the European corn borer (ECB, Ostrinia nubilalis), one of the most destructive pests of corn in North America and the western world. Our goals were to establish an ECB larval gut-specific EST database as a genomic resource for future research and to explore candidate genes potentially involved in insect-Bt interactions and Bt resistance in ECB. RESULTS: We constructed two cDNA libraries from the guts of the fifth-instar larvae of ECB and sequenced a total of 15,000 ESTs from these libraries. A total of 12,519 ESTs (83.4%) appeared to be high quality with an average length of 656 bp. These ESTs represented 2,895 unique sequences, including 1,738 singletons and 1,157 contigs. Among the unique sequences, 62.7% encoded putative proteins that shared significant sequence similarities (E-value

On-plant survival and inheritance of resistance to Cry1Ab toxin from Bacillus thuringiensis in a field-derived strain of European corn borer, Ostrinia nubilalis.

BACKGROUND: The high dose plus refuge is one of the major components of the resistance management plan mandated for transgenic corn expressing Cry toxins from Bacillus thuringiensis Berliner (Bt) that targets the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). This strategy was based on assumptions such as functional recessive inheritance, which has not been previously tested for O. nubilalis. The authors used a field-derived resistant strain of O. nubilalis to define the nature of resistance to Cry1Ab toxin by examining the inheritance and on-plant survival of susceptible and resistant insects and their F(1) progeny. RESULTS: The resistant strain exhibited > 800-fold resistance to Cry1Ab. Resistance was primarily autosomal and controlled by more than one locus or multiple alleles at one locus. The degree of dominance D calculated on the basis of LC(50) values was - 0.45(h' = 0.27), indicating that resistance was incompletely recessive. No survivors were found on vegetative-stage Bt corn, although both resistant larvae and their F(1) progeny were able to survive on reproductive corn 15 days after infestation. CONCLUSIONS: A field derived O. nubilalis strain exhibited high levels of resistance to Cry1Ab and survived on transgenic corn by feeding on tissues with low Cry1Ab expression. The Cry1Ab resistance was primarily autosomal, incompletely recessive and polygenic. Tissue and on-plant survival data indicated that dominance varies depending on plant stage.