Supplementary Figure S1 Separation and quantification of N-Formylloline (NFL) by gas chromatography (top) and its characterization by mass spectrometry (bottom). NFL elutes as a single peak at 7.67 minutes, while the phenylmorpholine standard elutes at 8.39 minutes. The peak at 6.85 minutes represents a contaminant on the column that eluted in all samples. While NFL is subject to considerable decomposition during mass spectrometry, the massspectra closely match that of authentic NFL as per Takeda et al. 1991.
Supplementary Figure S2 Separation and quantification of loline by gas chromatography (top) and its characterization by mass spectrometry (bottom). Loline elutes as a single peak at 7.58 minutes, while the phenylmorpholine standard elutes at 8.39 minutes. The peak at 6.85 minutes represents a contaminant on the column that eluted in all samples. While loline is subject to considerable decomposition during mass spectrometry, the massspectra obtained closely match that of authentic loline as per Takeda et al. 1991.
Supplementary Figure S3DNA sequences adjacent to the site of transposon insertion into mutants LCMS1 (top) and LCMS2 (bottom).
Supplementary Figure S4 The culturable tall fescue phyllosphere community before (top) and after (bottom) enrichment with lolines as the sole carbon source. Arrow indicate the distinct colonies types recovered after loline selection.
Supplementary Figure S5 Depletion of various loline species by growth of Burkholderia ambifaria 7R in a loline minimal medium as detected using thin layer chromatography. Shown are samples from an un-inoculated culture 72 hours after adding seed extract (left lane) and from cultures of B. ambifaria immediately after inoculation (center lane) and 72 hours after inoculation (right lane). The presence of loline (LOL), N-formylloline (NFL) and N-acetylloline (NAC) are shown with arrows. Note that the relative abundance of LOL to the other loline species is higher than determined directly by other means due to its preferential detection using drangendorff reagent used in TLC detection.
Supplementary Figure S6Growth of Burkholderia ambifaria7R withloline as a sole carbon source in a minimal medium (triangles) and depletion of this resource with time in the presence of cells (diamonds) or in cell free minimal medium (squares).
SupplementaryFigure S7 Lack of depletion of various loline species by growth of Pseudomonas fluorescens strain A506 in a loline minimal medium as detected using thin layer chromatography. Shown are samples from cultures incubated for various times after inoculation with bacteria from 0 to 144 hours (lanes from left to right). The presence of loline (LOL), N-formylloline (NFL) and N-acetylloline (NAC) are shown with arrows.
Supplementary Figure S8 Lack of depletion of various loline species by growth of Burkholderia ambifaria catabolic mutant LCMS1 in a loline minimal medium as detected using thin layer chromatography. Shown are samples from an un-inoculated culture 72 hours after adding seed extract (left lane) and from cultures of LCMS1 72 hours after inoculation (right lane). The presence of loline (LOL) and N-formylloline (NFL) are shown with arrows.
SupplementaryFigure S9 Concentration of cells of wild type Burkholderia ambifaria strain 7R and the loline catabolic deficient mutant LCMS1in minimal medium lacking a Carbon source (control) or in minimal medium containing glucose (Dextrose) or 200µg/ml N-formylloline (Loline) as a sole carbon source at either the time of inoculation (dark bars) 48 hours after inoculation (striped bars) or 192 hours after inoculation (white bars). The vertical lines represent the standard error of log cell concentration.
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