Figure 1.
The location and brief biosynthesis pathway of indoles, iridoids and MIAs.
Figure 2.
Representative PCR and Southern-blot analyses of independent transgenic C. roseus plants.
A: PCR analysis for the presence of ORCA3 in OR and GO plants; +: Positive control (pGO plasmid); W: wild type plant (negative control). B: PCR analysis for the presence of G10H in OR and GO plants; +: Positive control; W: wild type plants (negative control). C: Southern-blot analysis of transgenic C. roseus plants; +: Positive control (GO plasmid digested by SalI); WT: wild type plants.
Figure 3.
ORCA3, G10H, tdc, str and crmyc2 transcripts in OR and GO plants.
Error bars represent the standard deviation of triplicate runs for Real Time PCR. *: significant difference (p<0.05 by ANOVA).
Figure 4.
The average levels of several important MIAs in OR, GO and control lines.
The value of CK is the average of 15 control plants. Different letters indicate significant difference (p<0.05 by ANOVA). The value of OR is the average of pooled samples from 3 OR lines; the value of GO is the average of pooled samples from 5 GO lines.
Figure 5.
1H-NMR investigation and multivariate data analysis of GO and control lines.
A: 1H-NMR spectra of GO (in green) and control (in red) lines. B: Score plot of PLS-DA of GO (Δ) and control (▪) lines. C: Loading plot of PLS-DA. The numbers represent metabolites following table 1.
Figure 6.
Multivariate data analysis of OR and control lines based on 1H-NMR spectra.
A: PCA score plot of OR lines (OR) and control lines (W); B: PCA column plot of PC1. The numbers represent metabolites following table 1.
Table 1.
ANOVA results for selected signals from OR, GO transgenic and wild type plants.
Figure 7.
Schematic effects of ORCA3, or with G10H, overexpression on the metabolism of C. roseus plants.
Orange box is for ORCA3 overexpression (the OR lines) and blue box is for G10H and ORCA3 co-overexpression (the GO lines). The up arrow in the box represents the increase of metabolite content. The down arrow in the box represents the decrease of metabolite content. Arrows with star in the box represent significant difference (p<0.05 by ANOVA) of metabolite content compared with the controls.