Bacillus thuringiensisisolates from the Philippines: habitat distribution, δ-endotoxin diversity, and toxicity to rice stem borers (Lepidoptera: Pyralidae)

Abstract

AbstractBacillus thuringiensisBerliner isolates were detected in 57% of 801 samples of rice grain dust, soil, rice field arthropods, and miscellaneous habitats (rice straw compost and mammal faeces) collected at 100 sites in the Philippines. The collection yielded 3950 isolates ofB. thuringiensis(8.7 isolates/positive sample). Grain dust from rice mills was the richest source (63% of the samples were positive, with 10.2 isolates/positive sample), followed by rice field arthropods, soil, and miscellaneous habitats. Polyclonal antibodies to six δ-endotoxin groups (Cry1A, Cry1B, Cry1C, Cry1D, Cry1E, and Cry3A) were used in enzyme-linked immunosorbent assays (ELISA) to characterize the toxins produced by each isolate. Subsamples of isolates representing the diversity of isolate sources and δ-endotoxin profiles were bioassayed against the yellow stem borer,Scirpophaga incertulas(Walker) and striped stem borer,Chilo suppressalis(Walker). Eighteen isolates highly toxic to both species were selected for characterization of δ-endotoxin genes by polymerase chain reaction (PCR) with primers specific to 14 genes or gene subfamilies, and Western blotting with Cry2A antibodies. At least two novel δ-endotoxin genes, related tocry1Bandcry1F, were detected by DNA sequencing of PCR products.