Format

Send to

Choose Destination
Phytochemistry. 2014 Mar;99:73-85. doi: 10.1016/j.phytochem.2013.12.007. Epub 2014 Jan 25.

Metabolic flux phenotype of tobacco hairy roots engineered for increased geraniol production.

Author information

1
Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK.
2
VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT (Espoo), Finland.
3
Laboratory of Plant Physiology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
4
Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK. Electronic address: lee.sweetlove@plants.ox.ac.uk.

Abstract

The goal of this study was to characterise the metabolic flux phenotype of transgenic tobacco (Nicotiana tabacum) hairy roots engineered for increased biosynthesis of geraniol, an intermediate of the terpenoid indole alkaloid pathway. Steady state, stable isotope labelling was used to determine flux maps of central carbon metabolism for transgenic lines over-expressing (i) plastid-targeted geraniol synthase (pGES) from Valeriana officinalis, and (ii) pGES in combination with plastid-targeted geranyl pyrophosphate synthase from Arabidopsis thaliana (pGES+pGPPS), as well as for wild type and control-vector-transformed roots. Fluxes were constrained by the redistribution of label from [1-¹³C]-, [2-¹³C]- or [¹³C6]glucose into amino acids, sugars and organic acids at isotopic steady state, and by biomass output fluxes determined from the fractionation of [U-¹⁴C]glucose into insoluble polymers. No significant differences in growth and biomass composition were observed between the lines. The pGES line accumulated significant amounts of geraniol/geraniol glycosides (151±24 ng/mg dry weight) and the de novo synthesis of geraniol in pGES was confirmed by ¹³C labelling analysis. The pGES+pGPPS also accumulated geraniol and geraniol glycosides, but to lower levels than the pGES line. Although there was a distinct impact of the transgenes at the level of geraniol synthesis, other network fluxes were unaffected, reflecting the capacity of central metabolism to meet the relatively modest demand for increased precursors in the transgenic lines. It is concluded that re-engineering of the terpenoid indole alkaloid pathway will only require simultaneous manipulation of the steps producing the pathway precursors that originate in central metabolism in tissues engineered to produce at least an order of magnitude more geraniol than has been achieved so far.

KEYWORDS:

Flux analysis; Geraniol; Metabolic engineering; Terpenoid indole alkaloids

PMID:
24472392
DOI:
10.1016/j.phytochem.2013.12.007
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center