PCR products from diagnostic PCRs used to confirm deletion mutations in Burkholderia glumae and N-acyl homoserine lactone (AHL) signal production and toxoflavin production of deletion mutants.

(A) PCR products amplified from primers, TofI(H)F and TofI(H)R, to confirm the tofI deletion in LSUPB145. Template DNA for each lane is as follows: 1, pKKSacBΔtofI; 2, genomic DNA of B. glumae 336gr-1; and 3, genomic DNA of B. glumae LSUPB145. (B) PCR products amplified with primers, TofR(H)F and TofR(H)R, to confirm the tofR deletion in LSUPB169. Template DNA for each lane is as follows: 1, pKKSacBΔtofR; 2, genomic DNA of B. glumae 336gr-1; and 3, genomic DNA of B. glumae LSUPB169. (C) PCR products amplified with primers, TofI(H)F and TofR(H)R, to confirm the tofI-tofR deletion in LSUPB139. Template DNA for each lane is as follows: 1, pKKSacBΔtofIMR; 2, genomic DNA of B. glumae 336gr-1; and 3, genomic DNA of B. glumae LSUPB139. M indicates the 1 kb Plus DNA ladder (Invitrogen, Santa Clara, CA, USA) used as a marker. (D) Violacein production, shown as a purple pigment, by the biosensor, Chromobacterium violaceum CV026, in the presence of the culture extracts of the B. glumae strains, 336gr-1, LSUPB145, LSUPB169, and LSUPB139. Photo was taken 48 h after application of bacterial culture extracts on C. violaceum CV026 inoculated onto a LB agar plate. (E) Toxoflavin production, shown as a yellow pigment, in the LB broth by B. glumae strains, 336gr-1, LSUPB145, LSUPB169, and LSUPB139. Photo was taken after 24 h incubation at 37°C.