Early transcription of Bacillus thuringiensis cry genes in strains active on Lepidopteran species and the role of gene content on their expression

Abstract

Six strains of Bacillus thuringiensis previously selected as highly toxic against Manduca sexta and Plutella xylostella were analyzed by PCR screening in order to identify the cry genes active on Lepidoptera. According to their gene content and insecticidal potency, these strains were cultured and aliquots taken at different pre- and post-sporulation times. Total RNA was extracted and used as template in RT-PCR analyses directed to identify mRNAs of the previously identified cry genes. Results showed transcription of genes cry1A, cry1E, cry1I, and cry2 even before the onset of sporulation. However, this early transcription did not lead to an appreciable parasporal protein synthesis until t5-t9, as deduced from SDS-PAGE profiles. As for cry1I gene, the corresponding protein was not detected, as expected, but cry1I mRNAs were present at least until t5. Interestingly, strains expressing four cry genes from the end of the log phase onwards exhibited kinetics characterized by a very long transition phase, whereas the strain expressing only one cry gene showed a very short transition phase. Strains expressing three genes showed an intermediate profile. These results indicate that the transcription of B. thuringiensis cry1 and cry2 genes in natural strains can start several hours before massive crystal synthesis occurs and that this translation is probably competing with transcriptional regulators required for the sporulation onset.