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emi2867-sup-0001-si.doc5706K

Fig. S1. Biosynthesis of OLs in Agrobacterium tumefaciens. The genes coding for OlsB and OlsA have been first identified in Sinorhizobium meliloti. Here we describe the identification of the gene encoding the OL hydroxylase OlsE introducing a hydroxyl group in the ornithine moiety of OL S1 leading to the formation of OL S2. LOL, lyso-ornithine lipid.

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Fig. S2. Negative ion mode mass spectra of OL S1 (A) and OL S2 (B) purified from A. tumefaciens A208. The major OL [M-H] ion species are labelled with arrows.

emi2867-sup-0001-si.doc5706K

Fig. S3. Ornithine lipid S2 shows a different ninhydrin staining compared with OL S1. Unlabelled lipids were extracted according to Bligh and Dyer from 50 ml cultures of A. tumefaciens A208 (A) and R. tropici CIAT899 (B) grown in LB medium and separated in duplicate by two-dimensional TLC. Lipids were subjected to ninhydrin staining. The phospholipids PE, monomethyl PE (MMPE), and the OLs S1, S2, P1 and P2 are indicated.

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Table S1. Bacterial strains and plasmids used in this study.

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Table S2. Oligonucleotides used in this study. Introduced restriction sites are underlined.

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Table S3. Sequences used for PSSM construction using the program Info-Gibbs. Sequences were from taken from Yuan and colleagues (2006).

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