Metabolic compensation of steroidal glycoalkaloid biosynthesis in transgenic potato tubers: using reverse genetics to confirm the in vivo enzyme function of a steroidal alkaloid galactosyltransferase

摘要

Steroidal glycoalkaloids (SGAs) are secondary metabolites of Solanaceous plants. Two predominant glycoalkaloids, alpha-chaconine and alpha-solanine are produced in potatoes. An antisense transgene was constructed to down-regulate glycoalkaloid biosynthesis using a potato cDNA encoding a sterol alkaloid glycosyltransferase (Sgt1). Introduction of this construct into potatoes resulted in some lines with an almost complete inhibition of alpha-solanine accumulation. This inhibition was compensated by elevated levels of alpha-chaconine and resulted in wild type total SGA levels in the transgenic lines. In vitro assays with the recombinant SGT1 isolated from yeast demonstrated that Sgt1 encodes an enzyme capable of both glucosyltransferase and galactosyltransferase activity with a preference for UDP-galactose as the sugar donor. Together this data confirms SGT1's role in vivo as the solanidine:UDP-galactose galactosyltransferase. (C) 2004 Elsevier Ireland Ltd. All rights reserved.