Resveratrol Synthase Transgene Expression and Accumulation of Resveratrol Glycoside in Rehmannia glutinosa

Abstract

Rehmannia glutinosa L. is an important medicinal crop in Asian countries and contains trace amount of resveratrol compounds. To increase production of the compounds, we attempted ectopic expression of peanut resveratrol synthase gene (AhRS3) in R. glutinosa. The AhRS3 sequence that encompassed the open reading frame, including a 312 bp-long intron present between the 59th and 60th codon, was driven by the CaMV35S promoter and introduced into R. glutinosa via Agrobacterium-mediated transformation of leaf explants. The transgenic plants with one to three copies of AhRS3 transgene showed normal growth and development. The transgene was expressed constitutively in the leaf, root and flower at similar levels. Transgene expression in the leaf resulted in the production of new compounds identified as resveratrol and 3′-H-resveratrol-3-O-β-d-glucoside (R-gluc) through nuclear magnetic resonance spectroscopy and mass spectrometry. R-gluc accumulated predominantly and its content in the leaf of the 11 transgenic lines ranged from 22 to 116μg/gFW. The contents of resveratrol compounds in the transgenic plants were further increased by cold, UV, ethylene, and paraquat treatments, and were positively associated with the levels of AhRS3 mRNA levels. The R-gluc isolated from the transgenic plants exhibited antioxidant activity equivalent to one-third of resveratrol. Transgenic plants were highly resistant to Fusarium oxysporum infection. The results indicate that the ectopic expression of AhRS3 in R. glutinosa results in the production of R-gluc and resveratrol at hundreds of times higher levels than in peanut seed. The increased production of resveratrol compounds from R. glutinosa, which show diverse benefits for human and plant health, could provide a new opportunity for the improvement of R. glutinosa products.