title: Cryopreservation of in vitro-cultured multiple bud clusters of asparagus (Asparagus officinalis L. cv Hiroshimagreen (2n=30) by the techniques of vitrification.
abstract: A culture line of asparagus forming green bulbous structures consisting of numerous multiple bud clusters designated "bud clusters" was induced from a meristem culture of asparagus (Asparagus officinalis L.cv. Hiroshimagreen, 2n=30). Small cubic segments (2 mm3) cut from bud clusters were cryopreserved using three different cryogenic protocols. Only vitrification produced very high levels of shoot formation after cooling to -196°C. Segments were treated with a vitrification solution (PVS2) at 25°C for 45 min or at 0°C for 120 min prior to a direct plunge into liquid nitrogen. After rapid warming, the segments were expelled into Murashige and Skoog medium containing 1.2 M sucrose for 10 min and then plated on agar shoot outgrowth medium. The average rate of shoot formation of vitrified segments producing normal shoots was near 90% without any preculture and/or cold-acclimation treatment. Revived segments resumed growth within 3 days and developed about three shoots per segment. In vitro-cultured bud clusters appear promising as material for cryopreserving asparagus germplasm.
authors.affiliation: Institute of Biotechnology, Hiroshima Prefectural Agricultural Research Center, Hara, Hachihonmatsu, 739-01, Higashi-hiroshima, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kohmura
authors.forename: A
authors.initials: A
authors.lastname: Sakai
authors.forename: S
authors.initials: S
authors.lastname: Chokyu
authors.forename: T
authors.initials: T
authors.lastname: Yakuwa