Fig. 4.

A mutation in AT00_08765 leads to wrinkled morphology. a In-frame deletion of AT00_08765 was confirmed by PCR using four sets of primers. Lanes 1, 3, 5, and 7 used DNA from the wild-type strain, and lanes 2, 4, 6, and 8 used DNA from the deletion mutant. Lanes 1 and 2 were amplified using the primer pair 08765-SF/−SR, lanes 3 and 4 using 08765-SF/−LR, lanes 5 and 6 using 08765-LF/−SR, and lanes 7 and 8 using 08765-LF/−LR (Table S1). The expected product sizes were 1491, 2993, 2871, and 4373 bp for the wild-type and 382, 1884, 1762, and 3264 bp for Δ08765. b Swimming motility test of the Δ08765 strain versus the wild-type strain. c Mutant Δ08765 showed wrinkled morphology, whereas the wild-type strain showed smooth morphology in SW-LB medium. Complementation of a wild-type AT00_08765 via plasmid pBBR1MCS-08765 (p-08765) restored the colony morphology of Δ08765 and the phenotype of the wrinkled variant to smooth. d Congo red binding assay of the Δ08765 strain compared to the wild-type strain. At least two independent experiments of each strain were tested, and only representative images are shown