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Additional file 1: Table S1. Primers used for RT-PCR. Figure S1. The QsuB engineering strategy. Figure S2. Metabolic routes for microbial conversion of fermentable sugars from lignocellulosic hydrolysates into sesquiterpenes (epi-isozizaene and α-bisabolene) and fatty alcohols. Figure S3. Propagation and characterization of WT and QsuB poplar lines. Figure S4. Correlation between lignin content and DHB content in WT and QsuB biomass. Figure S5. Optical density of R. toruloides cultures after cultivation on large-scale poplar hydrolysates.

Additional file 1: Figure S1. Representative pictures showing GUS activities in various tiller sections of switchgrass lines harboring the pShOMT::GUS construct. GUS expression is specifically observed in stem nodes. Scale: White bars = 2 mm, black bar = 400 μ m. N: node; IN: internode, IS: internode transverse section.

Additional file 2: Figure S2. Characterization of a switchgrass line harboring the pZmCesa10::QsuB construct. (A) Representative pictures showing GUS activities in various tiller sections of switchgrass lines harboring the pZmCesa10::GUS construct. GUS expression is mostly observed in internodes, especially in developing vascular bundles (red arrows). Scale: White bars = 2 mm, black bar = 400 μ m. N: node; IN: internode, IS: internode transverse section. (B) Detection of the QsuB gene by PCR in line pZmCesa10::QsuB-5. (C) Detection of QsuB transcripts by RT-qPCR. QsuB expression levels relative to that of PvUBQ6 are shown. Values are means ±SD of two biological replicates (n = 2). (D) Protocatechuate (PCA) content measured in the biomass of the switchgrass line pZmCesa10::QsuB-5. A line containing the pZmCesa10::GUS construct was used as control. Values are means ±SE of three biological replicates (n = 3). Asterisks indicate a significant difference from the control using the unpaired Student’s t-test (*P

Additional file 4: Figure S3. Full length unprocessed images of PCR gels used for Figs. 2a and S2B. Note that the seven transformants obtained with the pZmUbi-1::QsuB construct were all false positives and did not contain the QsuB gene (see purple rectangle on the PCR gel).

Additional file 3: Table S1. Oligonucleotides used in the study.

Additional file 1: Supplemental Figures and Tables.

Additional file 1: Figure S1. Representative liquid chromatography���mass spectrometry (LC���MS) chromatograms obtained from analysis of TFA hydrolysates from CWR of wildtype (orange traces) and pSbUbi:AT10 (blue traces) sorghum stems. Peaks corresponding to pCA-Ara (a) and FA-Ara (b) display major ions at m/z 295 and 325, respectively. Electrospray ionization collision-induced dissociation tandem mass (ESI���CID���MS/MS) spectra of pCA-Ara (c) and FA-Ara (d) are shown. Ions at m/z 265 and 235 correspond to the 0,2A1 and 0,3A1 arabinose cross-ring cleavage ions as previously reported (Qu��m��ner and Ralet 2004). In addition, ions from deprotonated p-coumarate (m/z 163) and ferulate (m/z 193) are observed. Figure S2. Representative LC���MS chromatogram obtained from analysis of alkaline hydrolysates from CWR of wildtype sorghum stems. Seven peaks with ions at m/z 385 corresponding to diferulate isomers are observed. ESI���MS spectra from each peak are shown (bottom panels). Tentative diferulate chemical structures are based on the seven compounds previously identified in alkaline hydrolysates obtained from switchgrass and maize stalk cell walls (Ralph et al. 1994; Marita et al. 2003). Table S1. Growth parameters of wildtype and pSbUbi:AT10 lines. Values in brackets are the SE from five biological replicates (n = 5). Table S2. Characteristics and relative molar abundances (%) of guaiacyl (G) and syringyl (S) lignin-specific pyrolysis products released from CWR of wildtype (WT) and pSbUbi:AT10 lines. Values in brackets are the SE from four biological replicates (n = 4). Table S3. List of primers used in this study. Table S4. List of plasmids used in this study.