title: Conversion of exogenous cholesterol into glycoalkaloids in potato shoots, using two methods for sterol solubilisation.
abstract: Steroidal glycoalkaloids (SGA) are toxic secondary metabolites naturally occurring in the potato, as well as in certain other Solanaceous plant species, such as tomato, eggplant and pepper. To investigate the steroidal origin of SGA biosynthesis, cut potato shoots were fed cholesterol labelled with deuterium (D) in the sterol ring structure (D5- or D6-labelled), or side chain (D7-labelled), and analysed after three or five weeks. The labelled cholesterol and presence of D-labelled SGA were analysed by GC-MS and LC-MS/MS, respectively. When feeding D-labelled cholesterol solubilised in Tween-80, labelled cholesterol in free form became present in both leaves and stems, although the major part was recovered as steryl esters. Minor amounts of D-labelled SGA (α-solanine and α-chaconine) were identified in cholesterol-treated shoots, but not in blank controls, or in shoots fed D6-27-hydroxycholesterol. Solubilising the labelled cholesterol in methyl-β-cyclodextrin instead of Tween-80 increased the levels of labelled SGA up to 100-fold, and about 1 mole% of the labelled cholesterol was recovered as labelled SGA in potato leaves. Both side chain and ring structure D labels were retained in SGA, showing that the entire cholesterol molecule is converted to SGA. However, feeding side chain D7-labelled cholesterol resulted in D5-labelled SGA, indicating that two hydrogen atoms were released during formation of the SGA nitrogen-containing ring system. Feeding with D7-sitosterol did not produce any labelled SGA, indicating that cholesterol is a specific SGA precursor. In conclusion, we have demonstrated a superior performance of methyl-β-cyclodextrin for delivery of cholesterol in plant tissue feeding experiments, and given firm evidence for cholesterol as a specific sterol precursor of SGA in potato.
authors.affiliation: Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Centre for Plant Biology, Uppsala, Sweden.
authors.forename: Erik V
authors.initials: EV
authors.lastname: Petersson
authors.affiliation: Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Centre for Plant Biology, Uppsala, Sweden.
authors.forename: Nurun
authors.initials: N
authors.lastname: Nahar
authors.affiliation: Department of Ecology, Environment and Plant Sciences, Stockholm University, Stockholm, Sweden.
authors.forename: Paul
authors.initials: P
authors.lastname: Dahlin
authors.affiliation: Department of Chemistry, Uppsala BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Broberg
authors.affiliation: Chemistry Division 1, Science Department, National Food Agency, Uppsala, Sweden.
authors.forename: Rikard
authors.initials: R
authors.lastname: Tröger
authors.affiliation: Department of Food Science, Uppsala BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Paresh C
authors.initials: PC
authors.lastname: Dutta
authors.affiliation: Department of Ecology, Environment and Plant Sciences, Stockholm University, Stockholm, Sweden.
authors.forename: Lisbeth
authors.initials: L
authors.lastname: Jonsson
authors.affiliation: Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Centre for Plant Biology, Uppsala, Sweden.
authors.forename: Folke
authors.initials: F
authors.lastname: Sitbon
mesh_terms: D000470:Alkaloids / Q000096:biosynthesis*; D002784:Cholesterol / Q000378:metabolism / Q000494:pharmacology; D003903:Deuterium; D007553:Isotope Labeling / Q000379:methods; D018520:Plant Shoots / Q000378:metabolism*; D035281:Plant Tubers / Q000378:metabolism; D011198:Solanum tuberosum / Q000378:metabolism*
chemical_list: D000470:Alkaloids; D002784:Cholesterol; D003903:Deuterium