Assessing the limitations to terpenoid indole alkaloid biosynthesis in Catharanthus roseus hairy root cultures through gene expression profiling and precursor feeding
Assessing the limitations to terpenoid indole alkaloid biosynthesis in Catharanthus roseus hairy root cultures through gene expression profiling and precursor feeding
- Biotechnology Progress, 25(5), p.1289-1296, 2009 .
The production of pharmaceutically important terpenoid indole alkaloids (TIAs)from Catharanthus roseus is partly regulated at the transcriptional level. In this study, limitations in TIA biosynthesis from C. roseus hairy root cultures were assessed through gene expression profiling and precursor feeding. The transcript levels of key TIA pathway genes (G10h, Tdc, Str, and Sgd)and metabolite levels associated with the TIA pathway (tryptamine, loganin, secologanin, strictosidine, ajmalicine, serpentine, and tabersonine)were monitored using quantitative RT-PCR and HPLC, respectively. In cultures elicited with methyl jasmonate (250 M MeJA on day 21), G10h, Tdc, Str, and Sgd expression increased by 9.1, 3.1, 6.7, and 8.3-fold, respectively, after 24 h. Up-regulation of gene expression was followed by a 160, 440, and 420
CATHARANTHUS ROSEUS
HAIRY ROOT CULTURES
GENE EXPRESSION
METHYL JASMONATE
QUANTITATIVE RT-PCR
ALKALOID
PRECURSOR FEEDING
The production of pharmaceutically important terpenoid indole alkaloids (TIAs)from Catharanthus roseus is partly regulated at the transcriptional level. In this study, limitations in TIA biosynthesis from C. roseus hairy root cultures were assessed through gene expression profiling and precursor feeding. The transcript levels of key TIA pathway genes (G10h, Tdc, Str, and Sgd)and metabolite levels associated with the TIA pathway (tryptamine, loganin, secologanin, strictosidine, ajmalicine, serpentine, and tabersonine)were monitored using quantitative RT-PCR and HPLC, respectively. In cultures elicited with methyl jasmonate (250 M MeJA on day 21), G10h, Tdc, Str, and Sgd expression increased by 9.1, 3.1, 6.7, and 8.3-fold, respectively, after 24 h. Up-regulation of gene expression was followed by a 160, 440, and 420
CATHARANTHUS ROSEUS
HAIRY ROOT CULTURES
GENE EXPRESSION
METHYL JASMONATE
QUANTITATIVE RT-PCR
ALKALOID
PRECURSOR FEEDING