SEARCH

SEARCH BY CITATION

Figure S1. Western blots of CYP99A2, and CYP99A3, as indicated, following expression in Sf21 insect cell cultures as 6xHis tagged proteins, and as detected with an anti-His tag probe.

Figure S2. Optimized gene sequence for CYP99A2.

Figure S3. Optimized gene sequence for CYP99A3.

Figure S4. Summary of NMR structural analyses. (a) Carbon numbering for the syn-pimara-7(8),15(16)-dien-ol, al, or oic acid and syn-stemoden-19-ol,al,or oic acid diterpenoids. (b) Observed HMBC NMR spectra signals for syn-pimara-7(8),15(16)-dien-ol and syn-stemoden-19-ol. These are representative of the aldehydes and acids as well, minus the C19 proton correlations since C19 in the aldehydes and acids is quaternary. (c) Important cross-peak signals from NOESY spectra for the alcohols were used to determine which C4 geminal methyl group, C18 or C19, was the oxidized carbon. There was a strong signal between the alcohol protons and the C20 methyl group (known to be (R) beta from the syn configuration) protons for syn-pimara-7(8),15(16)-dien-ol and syn-stemoden-19-ol, indicating that C19 ((R) or beta on C4) is the oxidized carbon.

Figure S5. Kinetic plots for rCYP99A3 with (a) syn-pimaradiene and (b) syn-stemodene.

Figure S6. Detection of syn-pimaradien-19-oic acid, but not syn-stemoden-19-oic acid, from rice plant metabolites (both analyzed as methyl ester derivatives). Extracted GC-MS chromatogram of authentic standard, methyl jasmonate induced fractionated rice leaf extract (72 h time point), and uninduced (control) fractionated rice leaf extract for (a) syn-pimaradien-19-oic acid (m/z = 241), and (b) syn-stemoden-19-oic acid (m/z = 257).

Table S1.1H and 13C NMR assignments for syn-pimara-7,15-dien-19-ol (500 MHz at 25°C in chloroform-d).

Table S2.1H and 13C NMR assignments for syn-pimara-7,15-dien-19-al (700 MHz at 25°C in chloroform-d).

Table S3.1H and 13C NMR assignments for syn-pimara-7,15-dien-19-oic acid (500 MHz at 25°C in chloroform-d).

Table S4.1H and 13C NMR assignments for syn-stemod-13(17)-en-19-ol (500 MHz at 25°C in methanol-d4).

Table S5.1H and 13C NMR assignments for syn- stemod-13(17)-en-19-al (500 MHz at 25°C in methanol-d4).

Table S6.1H and 13C NMR assignments for syn- stemod-13(17)-en-19-oic acid (500 MHz at 25°C in methanol-d4).

As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer-reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.

FilenameFormatSizeDescription
TPJ_4408_sm_Legends.doc42KSupporting info item
TPJ_4408_sm_FigureS1.tif2933KSupporting info item
TPJ_4408_sm_FigureS2.doc34KSupporting info item
TPJ_4408_sm_FigureS3.doc34KSupporting info item
TPJ_4408_sm_FigureS4.eps321KSupporting info item
TPJ_4408_sm_FigureS5.eps1053KSupporting info item
TPJ_4408_sm_FigureS6.eps279KSupporting info item
TPJ_4408_sm_TablesS1-S6.doc150KSupporting info item

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.