Chen GuangyuConner, AJChristey, MCFautrier, AGField, RJ2008-05-141997-09Chen, G. Y., Conner, A. J., Christey, M. C., Fautrier, A. G., & Field, R. J. (1997). Protoplast isolation from shoots of asparagus cultures. International Journal of Plant Sciences, 158(5), 537-542.1058-5893YC995 (isidoc)https://hdl.handle.net/10182/477Experiments to maximize the isolation and purification of viable protoplasts from shoot cultures of asparagus (Asparagus officinalis L.) were conducted. Important factors for high yield of viable protoplasts included: the use of in vitro etiolated shoots as source material; 0.6 M glucose as an osmoticum in a modified KM medium; a combination of pectinase, cellulase, and hemicellulase, each at 1% (w/v) for enzymatic digestion of cell walls; and physical factors such as the volume of enzyme solution and speed of gyratory shaking. Protoplasts were purified by suspending digested etiolated shoot tissue in 0.6 M sucrose, overlaid with KMG medium and centrifugation at 650 g. The asparagus genotype had a marked influence on protoplast yield, with some genotypes yielding up to 18.4 x 10⁶ protoplasts/g fresh etiolated shoot tissue with 90% viability.pp.537-542enCopyright © 1997 The University of Chicago Presstissue culturein-vitro-cultureisolationprotoplastsshootsasparagusculturesJournal ArticleMarsden::270100 Biochemistry and Cell BiologyMarsden::270800 Biotechnology10.1086/2974641537-5315ANZSRC::3103 EcologyANZSRC::3104 Evolutionary biologyANZSRC::3108 Plant biology