Fig. S1 SDS-page of maize FDPS protein. Lane 1, purified FPPS3 in pASK-IBA37plus; lane 2, purified FPPS2 in pASK-IBA37plus; lane S1, unstained protein marker (Fermentas SM0669); lane 3, protein extract of FPPS3 in pASK-IBA37plus; lane 4, protein extract of FPPS2 in pASK-IBA37plus; lane 5, empty vector pASK-IBA37plus; lane S2, unstained protein marker (Thermo Scientific 26614). 12S1345S2 50kDA 20kDA A maize (Zea mays) herbivore-induced farnesyl diphosphate synthase supports the sesquiterpene synthesis in leaves. Annett Richter 1, Irmgard Seidl-Adams 2, Tobias Köllner 3, Joerg Degenhardt 1 1 Institute of Pharmacy, Martin Luther University, Hoher Weg 8, Halle, Germany, 2 Chemical Ecology, The Pennsylvania State University, State College PA 16802, 3 Max Planck Institute for Chemical Ecology, Hans-Knöll-Strasse 8, Jena, Germany

FPPS2+ IDP+ DMADP Farnesol a FPPS3+ IDP+ DMADP Farnesol b Fig. S2 Activity assay of FPPS after heterologous expression in E. coli. FPPS2 (a) and FPPS3 (b) without His-tag and the empty vector control (d) were incubated with the substrates IDP and DMADP for 45 min. The assay in panel c was incubated with both recombinant FPPS3 and TPS23, a maize (E)-ß-caryophyllene synthase that converts the FDP of FPPS3 to (E)-ß- caryophyllene. To the assays in panel a, b and d, hydrochloric acid was added to convert the prenyl phosphate product FDP into the corresponding alcohol farnesol. The volatiles were detected by GC-MS. Retention time (min) empty vector+ IDP+ DMADP Farnesol Relative abundance(TICx 10 7 ) FPPS3+ IDP + DMADP+ TPS (E)-ß-caryophyllene c d

Transcript level relative to APT1 Spod. litt. treated Spod. litt. treated Spod. litt. treated Spod. litt. treated Spod. litt. treated Fig. S3 Transcript abundance of fpps1, fpps2, and fpps3 genes in leaves after attack by Spodoptera littoralis. Transcript levels were determined in leaves of 14 day-old plants of the variety Delprim (a) and B73 (b) that were undamaged, wounded mechanically, or treated with third instar larvae of S. littoralis. for 24h. The expression was calculated relative to the housekeeping gene APT1. Significance was calculated by one-way-ANOVA of three technical repeats of four biological samples. Significant differences among treatments for each gene for P≤ (***). ***

Transcript level relative to APT1 *** Fig. S4 Transcript abundance of fpps1, fpps2, and fpps3 genes in leaves. Transcript levels were determined in leaves of 14 day-old plants of the inbred line B73 that were undamaged, wounded mechanically, and wounded and treated with elicitor (indanone-derivate) for 24h. The expression was calculated relative to the housekeeping gene APT1. Significance was calculated by one-way-ANOVA of three technical repeats of four biological samples. Significant differences among treatments for each gene were indicated for P≤ (***). FPPS2 FPPS1

Transcript level relative to APT1 tps23 Fig. S5 Transcript abundance of tps23 in leaves. Transcript levels were determined in leaves of 14 day-old plants of the variety Delprim that were undamaged, wounded mechanically, and wounded and treated with elicitor (indanone-derivate) for 24h. The expression was calculated relative to the housekeeping gene APT1. Significance was calculated by one-way-ANOVA of three technical repeats of four biological samples. Significant differences were indicated for P≤ (***). ***

GeneNameSequenceApplication GRMZM2G131907APT1_FAGGCGTTCCGTGACACCATCAPT1 QRT fwd APT1_R CTGGCAACTTCTTCGGCTTCCAPT1 QRT rev GRMZM2G147721FPPS3FCCTGGCTAGTTGTGCAAGCTFPPS2 QRT fwd FPPS8RGAAAACAcTCTGGACTGCCTFPPS2 QRT rev FPPS2_RGTCGACGGAGCAGCTTCTTCFPPS2 cloning fwd FPPS2_pASK-IBA33_FATGGTAGGTCTCAAATGGCGACGGTGGAGGTGGTGGT FPPS2 cloning (pASK- IBA33+) fwd FPPS2_pASK-IBA33_RATGGTAGGTCTCAGCGCTCTTGTCCCTCTTGTAGATCTTGTG FPPS2 cloning (pASK- IBA33+) rev FPPS2_pASK-IBA37_FATGGTAGGTCTCAGCGCATGGCGACGGTGGAGGTGGTG FPPS2 cloning (pASK- IBA37+) fwd FPPS2_pASK-IBA37_RATGGTAGGTCTCATATCACTTGTCCCTCTTGTAGATCTTGTG FPPS2 cloning (pASK- IBA37+) rev FPPS2_FCGAGGCCGCAATGGCGACGGTFPPS2 cloning rev GRMZM2G168681FPPS8.1FCATGGCTAGTTGTGCAAGCCFPPS1 QRT fwd FPPS8.1RCAGCACTTTCTGAACCGCAAFPPS1 QRT rev GRMZM2G098569FPPS3FCCTGGCTAGTTGTGCAAGCTFPPS3 QRT fwd FPPS3RGAAAACAGTTTGGACTGCCTFPPS3 QRT rev FPPS3_F CGAGGCCGCAATGGCGACAGCFPPS3 cloning fwd FPPS3_R GTCGATGGAGCAGCTTGTCGFPPS3 cloning rev FPPS3_pASK-IBA33_FATGGTAACCTGCATTAAATGGCGACAGCGGAGGTGGTGGT FPPS3 cloning (pASK- IBA33+) fwd FPPS3_pASK-IBA33_RATGGTAACCTGCATTAGCGCTCTTGTCCCTCTTGTAGATCTTGTG FPPS3 cloning (pASK- IBA33+) rev FPPS3_pASK-IBA37_FATGGTAACCTGCATTAGCGCATGGCGACAGCGGAGGTGGTG FPPS3 cloning (pASK- IBA37+) fwd FPPS3_pASK-IBA37_RATGGTAACCTGCATTATATCACTTGTCCCTCTTGTAGATCTTGTG FPPS3 cloning (pASK- IBA37+) rev Table S1 Primer for QRT-PCR, isolation of open reading frames and cloning into expression vectors of fpps1, fpps2 and fpps3

Plant speciesPrenyldiphosphate synthase Accession number Zea maysZmFPPS3BT ZmFPPS1BT ZmFPPS2LOC ZmGGPPS1EF zmGGPPS2EF ZmGGPPS3EF Oryza sativaOsFPPS1D85317 OsFPPS2LOC_Os05g46580 OsFPPS3LOC_Os04g56230 A.thalianaAtFPS1NM_ AtFPS2LNM_ AtGGPPS2U44876 AtGGPPS4AC Table S2 Accession numbers of the rooted phylogenetic tree of FPPSs and GGPPs of different plant species