Abstract

Rutin, one of the metabolites of the flavonoid pathway, shows great potential in industrial applications as a key component in pharmaceutical medicines and biological pesticides. Although the genetic manipulation of transcription factors (TFs) could increase rutin levels in plants, the accompanying accumulation of structurally similar chemicals complicates industrial rutin extraction. In this study, we demonstrated remarkably elevated rutin content (3.5-4.4-fold relative to controls) in transgenic tobacco plants by overexpressing NtFLS2. The levels of other intermediates in the branch pathway did not change much except for a moderate increase of kaempferol-3-O-rutinoside. Furthermore, the transcript levels of pathway genes in transgenic lines were comparable with controls, indicating genetic engineering did not significantly alter the branch pathway. Additionally, the transgenic tobacco plants appeared normal except for a flower color change from light red to white suggesting that it could be a valuable material for industrial extraction of rutin.

Highlights

  • Rutin, one of the metabolites of the flavonoid pathway, shows great potential in industrial applications as a key component in pharmaceutical medicines and biological pesticides

  • The overexpression of flavonol-specific transcription factors has been demonstrated to enhance the rutin accumulation in the tobacco leaf and leads to significant changes in flavonoid profiles.10,11,15) This may result in the production of structurally similar metabolites and complicate the purification process due to the structural similarity of other byproducts with rutin

  • It was reported that the silencing of FLS leads to fruits with arrested seed set and reduced quercetin content.16) no studies have reported on the influence of FLS on rutin biosynthesis

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Summary

Introduction

One of the metabolites of the flavonoid pathway, shows great potential in industrial applications as a key component in pharmaceutical medicines and biological pesticides. The genetic manipulation of transcription factors (TFs) could increase rutin levels in plants, the accompanying accumulation of structurally similar chemicals complicates industrial rutin extraction. We demonstrated remarkably elevated rutin content (3.5-4.4-fold relative to controls) in transgenic tobacco plants by overexpressing NtFLS2. A glycosylated flavonoid, is derived from dihydroquercetin by sequential reactions catalyzed by flavonol synthase (FLS), glucosyltransferase (GT), and rhamnosyltransferase (RT) (Supplementary Fig. 1). Tomato and tobacco plants with enhanced rutin content have been achieved via the heterologous expression of transcription factor AtMYB12 from Arabidopsis.10,11) A subsequent study on calluses from AtMYB12-expressing tobacco showed higher rutin levels than controls.12) In addition, overexpression of AtMYB11, a homolog of AtMYB12 significantly increased the rutin content in leaf of tobacco and tomato.). Transgenic plants overexpressing NtFLS2 were generated to investigate its function in rutin biosynthesis

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