title: Cryopreservation of the australian species Macropidia fuliginosa (Haemodoraceae) by vitrification.
abstract: Somatic embryos were used to develop a cryopreservation protocol for Macropidia fuliginosa, a commercially-important species endemic to the south-west of Western Australia. Somatic embryos were allowed to develop from embryogenic callus for three weeks on an kinetin medium prior to processing. These were transferred and cultured on a agar solidified basal medium supplemented with 0 to 0.6 M sorbitol for 2 d prior to incubation in Plant Vitrification Solution Two (PVS2). Following this, embryos were then washed in 1 M sucrose solution (treated controls) or cooled in liquid nitrogen (LN). Cooled embryos were then warmed and washed in sucrose solution. Highest survival for cooled treatments (67.3%) was achieved by preculture with 0.4 M sorbitol, then incubation in PVS2. Further experimentation varying pre-culture duration (2 or 3 d) and incubation on either glycerol (0.8 M) or sorbitol (0.4 M) indicated that very high survival (90.6%) of embryos was achievable by adopting a 2 d preculture period on 0.8 M glycerol. The phenotype and growth rates of plants obtained using this protocol were similar to those of parent plants. This optimised procedure was then applied to tissue culture-derived shoot apices of the same clone also resulting in a high survival rate (84.4%).
authors.affiliation: Kings Park and Botanic Garden, West Perth, WA, 6005, Australia. Curtin University of Technology, Bentley, WA, 6102, Australia. turnersr@pop.ses.curtin.edu.au
authors.forename: S R
authors.initials: SR
authors.lastname: Turner
authors.forename: B
authors.initials: B
authors.lastname: Tan
authors.forename: T
authors.initials: T
authors.lastname: Senaratna
authors.forename: E
authors.initials: E
authors.lastname: Bunn
authors.forename: K W
authors.initials: KW
authors.lastname: Dixon
authors.forename: D H
authors.initials: DH
authors.lastname: Touchell