(+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways

K. Cankar, E.J. Jongedijk, M. Klompmaker, T. Majdic, R. Mumm, H.J. Bouwmeester, H.J. Bosch, M.J. Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

31 Citations (Scopus)


Plant sesquiterpenes, such as (+)-valencene, artemisinin, and farnesene are valuable chemicals for use as aromatics, pharmaceuticals, and biofuels. Plant-based production systems for terpenoids critically depend on the availability of farnesyl diphosphate (FPP). Currently, these systems show insufficient yields, due to the competition for FPP of newly introduced pathways with endogenous ones. In this study, for the first time an RNAi strategy aiming at silencing of endogenous pathways for increased (+)-valencene production was employed. Firstly, a transient production system for (+)-valencene in Nicotiana benthamiana was set up using agroinfiltration. Secondly, silencing of the endogenous 5-epi-aristolochene synthase (EAS) and squalene synthase (SQS) that compete for the FPP pool was deployed. This resulted in a N. benthamiana plant that produces (+)-valencene as a prevalent volatile with a 2.8-fold increased yield. Finally, the size of the FPP pool was increased by overexpression of enzymes that are rate-limiting in FPP biosynthesis. Combined with silencing of EAS and SQS, no further increase of (+)-valencene production was observed, but emission of farnesol. Formation of farnesol, which is a breakdown product of FPP, indicates that overproducing sesquiterpenes is no longer limited by FPP availability in the cytosol. This study shows that metabolic engineering of plants can effectively be used for increased production of desired products in plants. Keywords: 5-Epi-aristolochene synthase · Metabolic engineering · RNAi · Squalene synthase
Original languageEnglish
Pages (from-to)180-189
JournalBiotechnology Journal
Issue number1
Publication statusPublished - 2015


  • plant transformation
  • biosynthetic-pathway
  • terpenoid metabolism
  • squalene synthase
  • tobacco
  • expression
  • artemisinin
  • arabidopsis
  • reductase
  • precursors


Dive into the research topics of '(+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways'. Together they form a unique fingerprint.

Cite this