title: The isolation and culture of protoplasts from an embryogenic cell suspension culture of Picea glauca (moench) voss.
abstract: Conditions were standardized for the isolation and culture of protoplasts from an embryogenic cell suspension culture of Picea glauca. A combination of 0.5% Cellulase R-10, 0.25% Macerozyme, 0.25% Driselase, 0.25% Rhozyme HP-150 with 0.5M mannitol and 5 mM CaCl2.2H2O produced an average of 4.5 × 10(6) protoplasts per gram fresh weight of cells. Of the several protoplast culture media tested, von Arnold and Eriksson and Kao and Michayluk (KM8P) media best supported mitotic divisions of protoplasts. A density of 10(5) protoplasts per ml and the addition of 5 mM glutamine to the culture medium was necessary to induce sustained divisions and microcallus formation. Microcalli grew into subculturable callus using a nurse culture technique.
authors.affiliation: Plant Biotechnology Institute, National Research Council, 110 Gymnasium Road, S7N 0W9, Saskatoon, SK, Canada.
authors.forename: F
authors.initials: F
authors.lastname: Bekkaoui
authors.forename: P K
authors.initials: PK
authors.lastname: Saxena
authors.forename: S M
authors.initials: SM
authors.lastname: Attree
authors.forename: L C
authors.initials: LC
authors.lastname: Fowke
authors.forename: D I
authors.initials: DI
authors.lastname: Dunstan