Supplementary Table 1 Volatile compounds detected from green hairy roots of D. carota
Peak No. / Name of Compounds / RIa / IDb1 / 6-Methyl-5-hepten-2-one / 987 / MS,RI
2. / β-Myrcene / 991 / MS,RI
3 / Internal standard
4 / Octanal / 1005 / MS,RI
5. / p-Cymene / 1024 / MS,RI
6. / Limonene / 1028 / MS,RI
7. / γ-Terpinene / 1058 / MS,RI
8. / Undecane / 1100 / MS,RI
9. / Nonanal / 1104 / MS,RI
10. / Naphthalene / 1180 / MS,RI
11. / Methylsalicylate / 1193 / MS,RI
12 / Dodecane / 1200 / MS,RI
13. / Decanal / 1205 / MS,RI
14 / Geraniol / 1257 / MS,RI
15 / Geranial / 1270 / MS,RI
16 / Bornyl acetate / 1284 / MS,RI
17. / Tridacane / 1300 / MS,RI
18. / Undecanal / 1307 / MS,RI
19 / Geranyl acetate / 1383 / MS,RI
20 / Tetradecane / 1400 / MS,RI
21 / Dodecanal / 1409 / MS,RI
22 / β-Caryophyllene / 1419 / MS,RI
23 / trans-α- Bergamotene / 1436 / MS,RI
24 / cis-Geranylacetone / 1452 / MS,RI
25 / (E) β -Farnesene / 1457 / MS,RI
26 / β-Acoradiene / 1476 / MS,RI
27 / Pentadecane / 1500 / MS,RI
28 / β -Bisabolene / 1510 / MS,RI
29 / Geranyl isobutyrate / 1512 / MS,RI
30 / (3E,6E) α-Farnesene / 1516 / MS,RI
31 / Unknown sesquiterpene / 1533 / MS,RI
32 / Geranyl isovalerate / 1602 / MS,RI
aRI: Retention index in reference to normal alkanes (C8-C20)
bID: Identification was done by comparing mass spectrum (MS) of the compound with mass spectral library from NIST 05 (National Institute of Standards and Technology, Gaithersburg, USA) and Wiley 8.0 (Wiley, New York, USA); RI, identification by comparison of Retention index with the published literatures (da Silva et al. 1999; Daferera et al. 2003; Asuming et al. 2005)
Supplementary Fig. S1 In-gel SKDH activity of extracted total proteins from green (G) and normal hairy roots of D. carota. Crude protein extracts (150 μg) were separated in 10% native polyacrylamide gel and stained according to Díaz et al. (1997). Hairy root cultures of active growth phase (15-d after subculture) from both green and normal types were used in this study.
Supplementary Fig. S2 Amino acid sequence alignment of putative DXR (query) of D. carota and corresponding sequences from other plants. Genebank accession numbers are as follows: O. sativa(Oryza, AAL37560), Z. mays(Zea,ACG33012),A. thaliana (Arabidopsis, NP_001190600), R.verticillata(Rauvolfia,AAY87151),W. somnifera (Withania,AFI98879), and M.xpiperita (Mentha, AAD24768)
Supplementary Fig. S3PCR cycle normalization during expression analysis of DXR. a cDNA, synthesized from total RNA was amplified by different PCR cycle (20-35 cycle) and was run in agarose gel. b Relative density of bands appeared in gel after PCR amplification with different PCR cycle
Supplementary Fig. S4 GC-MS chromatogram of volatile compounds from green hairy roots of Daucus carota
Supplementary Fig. S5 Comparison of emission of methylsalicylate glyphosate treated and untreated green and normal hairy roots. Aqueous glyphosate (50 μM) solutions were used to inhibit phenolic biosynthesis in green and normal hairy roots. (a) Methylsalicylate content in untreated and treated green hairy roots. (b) Methylsalicylate content in untreated and treated normal hairy roots
Supplementary Fig. S6 Comparison of H2O2 content in green and normal hairy roots of D. carota. H2O2 content was measured spectrophotometrically by following method of Loreto and Velikova (2001). Crude extracts from both types of hairy roots were used to quantify H2O2in presence of KI solutions.