title: Synthesis of Glutathione in Leaves of Transgenic Poplar Overexpressing [gamma]-Glutamylcysteine Synthetase.
abstract: Internode stem fragments of the poplar hybrid Populus tremula x Populus alba were transformed with a bacterial gene (gshl) for [gamma]-glutamylcysteine synthetase ([gamma]-ECS) targeted to the cytosol. Lines overexpressing [gamma]-ECS were identified by northern analysis, and the transformant with the highest enzyme activity was used to investigate the control of glutathione synthesis. Whereas foliar [gamma]-ECS activity was below the limit of detection in untransformed plants, activities of up to 8.7 nmol mg-1 protein min-1 were found in the transformant, in which the foliar contents of [gamma]-glutamylcysteine ([gamma]-EC) and glutathione were increased approximately 10- and 3-fold, respectively, without affecting either the reduction state of the glutathione pool or the foliar cysteine content. A supply of exogenous cysteine to leaf discs increased the glutathione content from both transformed and untransformed poplars, and caused the [gamma]-EC content of the transformant discs to increase still further. The following conclusions are drawn: (a) the native [gamma]-ECS of untransformed poplars exists in quantities that are limiting for foliar glutathione synthesis; (b) foliar glutathione synthesis in untransformed poplars is limited by cysteine availability; (c) in the transformant interactions between glutathione synthesis and cysteine synthesis operate to sustain the increased formation of [gamma]-EC and glutathione; and (d) the foliar glutathione content of the transformant is restricted by cysteine availability and by the activity of glutathione synthetase.
authors.affiliation: Laboratoire du Metabolisme (G.N.) and Laboratoire de Biologie Cellulaire (L.J.), Institut National de la Recherche Agronomique, Route de Saint-Cyr, 78026 Versailles Cedex, France.
authors.forename: G.
authors.initials: G
authors.lastname: Noctor
authors.forename: M.
authors.initials: M
authors.lastname: Strohm
authors.forename: L.
authors.initials: L
authors.lastname: Jouanin
authors.forename: K. J.
authors.initials: KJ
authors.lastname: Kunert
authors.forename: C. H.
authors.initials: CH
authors.lastname: Foyer
authors.forename: H.
authors.initials: H
authors.lastname: Rennenberg