Abstract
Stevia (Stevia rebaudiana Bertoni) produces steviol glycosides (SG), a group of diterpenoid secondary metabolites widely used as natural sweeteners. SGs share the same precursor with gibberellins (GAs), ent-kaurenoic acid. In this study, we attempted to increase the SG content in stevia propagated in Temporary Immersion Bioreactors (TIBs) by blocking ent-kaurenoic acid entry to the GA synthesis pathway using Daminozide (0, 10, 20 ppm), a known GA inhibitor. The maximum increase in biomass weight and SG content (up to 40% for stevioside and rebaudioside A) was observed following 10 ppm Daminozide treatment. The treatment also increased transcript accumulation for genes involved in SG biosynthesis, such as SrKA13H, SrUGT85C2, and SrUGT76G1, indicating that the SG pathway became more active due to GA pathway inhibition. Furthermore, GA inhibition following inhibitor exposure was also indicated by the upregulated expression of a GA biosynthesis gene (GA3ox) due to feedback regulation, and the downregulated expression of a GA catabolism gene (GA2ox2) as a result of feed-forward regulation.
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