Electronic Supplementary Material (Plant Molecular Biology)

Electronic Supplementary Material (Plant Molecular Biology)

Suppression of CCR impacts on metabolite profile and cell wall composition in Pinus radiata tracheary elements

Armin Wagner1, Yuki Tobimatsu 2, GeertGoeminne4,5, LorellePhillips1, Heather Flint1, Diane Steward1, Kirk Torr1, Lloyd Donaldson1, WoutBoerjan4,5, John Ralph2,3

Corresponding author: Armin Wagner, Scion, Private Bag 3020, Rotorua, New Zealand, Phone: +64-7-343-5449, Fax: +64-7-343-5444, e-mail:

Online resource 1Pyrograms (total ion chromatograms) from purified TEs from a wild-type control (A) and CCR-deficient lines pHF1-35 (B), pHF1-40 (C) and pHF1-45 (D). Numbers 1-25 refer to the following pyrolysis products. 1: guaiacol; 2: 4-hydroxy-5,6-dihydro-(2H)-pyran-2-one; 3: 4-methyl-guaiacol; 4: phenol; 5: dimethyl-phenol; 6: eugenol; 7: 4-vinyl-guaiacol; 8: cis-isoeugenol; 9: trans-isoeugenol; 10: unknown; 1-(4-hydroxy-3-methoxyphenyl)propyne?; 11: unknown; 1-(4-hydroxy-3-methoxyphenyl)allene?; 12: vanillin; 13: 1,5-anhydro-arabinofuranose; 14: acetoguaiacone; 15: guaiacyl acetone; 16: catechol; 17: 4-methyl catechol; 18: 1,5-anhydro-β-D-xylofuranose; 19: dihydroconiferyl alcohol; 20: 1,6-anhydro-α-D-galactopyranose; 21: cis-coniferyl alcohol; 22: 1,6-anhydro-β-D-mannopyranose; 23: trans-coniferyl alcohol; 24: 1,6-anhydro-β-D-glucopyranose; 25: unknown. Lignin-derived signals (1, 3, 4, 5, 7, 8, 9, 12, 14, 15, 19, 21 and 23) are diminished and polysaccharide-derived signals (13, 18, 20, 22 and 25) elevated in transgenic lines compared to the wild-type control.

Online resource 2Pyrograms (total ion chromatograms) of purified TEs from CCR-deficient line pHF1-35 without (A) and after treatment with 1 mM (B) and 10 mM NaOH (C). Numbers 1-25 refer to the following pyrolysis products. 1: guaiacol; 2: 4-hydroxy-5,6-dihydro-(2H)-pyran-2-one; 3: 4-methyl-guaiacol; 4: phenol; 5: dimethyl-phenol; 6: eugenol; 7: 4-vinyl-guaiacol; 8: cis-isoeugenol; 9: trans-isoeugenol; 10: unknown; 1-(4-hydroxy-3-methoxyphenyl)propyne?; 11: unknown; 1-(4-hydroxy-3-methoxyphenyl)allene?; 12: vanillin; 13: 1,5-anhydro-arabinofuranose; 14: acetoguaiacone; 15: guaiacyl acetone; 16: catechol; 17: 4-methyl catechol; 18: 1,5-anhydro-β-D-xylofuranose; 19: dihydroconiferyl alcohol; 20: 1,6-anhydro-α-D-galactopyranose; 21: cis-coniferyl alcohol; 22: 1,6-anhydro-β-D-mannopyranose; 23: trans-coniferyl alcohol; 24: 1,6-anhydro-β-D-glucopyranose; 25: unknown. Polysaccharide-derived signals (13, 18, 20, 22 and 25) declined after NaOH treatment of TEs, which resulted in a stronger appearance of lignin-derived signals (1, 3, 4, 5, 7, 8, 9, 12, 14, 15, 19, 21 and 23) in pyrograms.

Online resource 3 Potential metabolic route for the formation of a phenolic compound in CCR-RNAilines that breaks down to vanillyl acetone during pyrolysis. It is speculated that feruloyl-CoA participates in the formation of the unknown phenolic compound based on the position of CCR in the phenylpropanoid pathway and the molecular structure of the pyrolysis product vanillyl acetone. PAL: phenylalanine ammonia-lyase; C4H: cinnamate 4-hydroxylase; 4CL: 4-coumarate-CoA ligase; HCT: p-hydroxycinnamoyl-CoA:shikimatehydroxycinnamoyl transferase; C3H: p-coumarate 3-hydroxylase; CCoAOMT: caffeoyl-CoAO-methyltransferase; CCR: cinnamoyl-CoA reductase; R: unknown residue; ?: unknown enzymatic steps potentially involving a double bond reductase (Kasahara et al., 2006).