Fig. 2: Regulation of TRI6 expression by TRI10 and vice versa.
a Relative expression levels of TRI10 in 3-day-old LTB cultures of the wild-type strain PH-1 (arbitrarily set to 1) and tri6 mutant were assayed by qRT-PCR with primers amplified the 3’-terminal region of TRI10. b Relative expression levels of TRI6 and antisense-TRI6 in PH-1 (arbitrarily set to 1) and the tri10 mutant were assayed with strand-specific qRT-PCR. Deletion of TRI10 significantly increased the transcription of antisense-TRI6. c Schematic drawing of the TRI6 ORF and its sense or antisense transcripts. In the TRI6AD mutant allele, the 185-bp promoter region (red box) was replaced with the hygromycin phosphotransferase (hph) fusion with thymidine kinase (TK) genes. TSS/TTS, transcription start/termination site. Fragments were amplified with the primer pairs T6Y-1F/ T6Y-2R and T6Y-3F/T6Y-4R for the generation of the TRI6AD strain, and the primer pair T6RTF/T6RTR was used for qRT-PCR assay. d Relative expression levels of TRI6 and antisense-TRI6 in 3-day-old LTB cultures of PH-1 and TRI6AD mutant. e DON production in 7-day-old LTB cultures of PH-1 and the TRI6AD mutants. For a, b, d and e, mean and standard deviation were estimated with data from three (n = 3) independent replicates (marked with black dots on the bars). For DON production, the statistical difference relative to PH-1 is based on the two-tailed unpaired t test (****, p < 0.0001). The exact p-values are shown in the Source Data file.
