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Fig. S1. Genomic organization of rfb genes in H. seropedicae. The arrows indicate the location of transposon (EZ:Tn5<TET-1>) insertion, generating the mutant strains H. seropedicae RAMEB-B (rfbB-) and RAMEB-C (rfbC-). *Orthologue of rfbA. Values expressed in number of base pairs.

Fig. S2. Motility of the H. seropedicae strains in NFbHPN 0.5% agar. A total of 105 cells of H. seropedicae strains were inoculated into the centre of the glass tube. The culture was incubated at 30°C and the movement of the cells down the glass tube and formation of surface biofilm in the surrounding medium were monitored over time. The wild-type (A) and mutant (B) strains showed the same motility pattern.

Fig. S3. A. Herbaspirillum seropedicae wild-type (1), rfbB (2) and rfbC (3) mutant strains grown in NFbHPN medium containing 0.02% calcofluor. B. Same as (A) irradiated by UV (365 nm) light.

Fig. S4. A. LPS profile from H. seropedicae grown in NFbHPN medium in the absence (lane 1) or presence of 5 mM CaCl2 (lane 2). SDS-PAGE was performed with total LPS extracted from wild-type cells. The densitometric analyses of the gel regions delimited by squares are plotted in (B). Red and green traces indicate the profile of H. seropedicae grown in the absence or in the presence of 5 mM CaCl2 respectively; arrows indicate bands containing LPS with higher number of repetitive oligosaccharide chains of the O-antigen which are more abundant when grown in the presence of Ca2+. For the analyses of LPS, H. seropedicae was cultivated for 16 h in liquid medium in the absence or presence of CaCl2 (5 mM). Total LPS was extracted from 107 cells. Equal amounts of sample were then separated by SDS-PAGE (16%) and visualized after silver periodate oxidation staining. The gel images were analysed with the LabWorks Imaging and Analysis Software (UVP, Cambridge, UK). The gel shown is representative of three independent experiments.

Table S1. Sequence similarities between proposed Rfb proteins from H. seropedicae and other dTDP-L-Rha biosynthetic enzymes.

Table S2. Monosaccharide composition of H. seropedicae strains LPS.

Table S3. Attachment of H. seropedicae strains to glass fibre.

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