Plant regeneration from callus-derived protoplasts of asparagus
1989
Elmer, W.H. | Ball, T. | Volokita, M. | Stephens, C.T. | Sink, K.C.
Donor callus cells for protoplasts were initiated from mature plants of four selected crowns of asparagus (Asparagus officinalis L.) by placing spear slices on solidified Murashige and Skoog salts and vitamins medium (MS) with 3% sucrose + (in mg.liter-1): 1.0 NAA + 1.2 2,4-D + 0.9 BA or 1.0 kinetin + 2.5 2,4-D. Callus derived from these explants was further subcultured on the same medium. Optimum protoplast yields were enzymatically obtained from such calluses 10 to 20 days after subculture. Of the isolated protoplasts 65% to 75% were viable, and when plated in modified Kao and Michayluk medium at 5 X 10(4) or 10(5)/ml densities, had 6.5% and 7.3% plating efficiencies, respectively. Protoplast isolations had 0.81% to 1.4% cells present that were not observed subsequently to undergo division. Only the cells or protoplasts of J̀ersey Giant crown No. 8' divided during 8 weeks to form microcalluses. After transfer and culture for an additional 4 to 5 weeks on solidified MS + (in mg.liter-1): 0.1 NAA + 1.0 kinetin, shoots regenerated at 28% efficiency. Shoots were rooted at 50% efficiency on solidified MS + (in mg.liter-1): 0.3 NAA + 0.7 kinetin + 2.1 ancymidol + 4% sucrose. The rooted plants were readily transferred to the greenhouse. Chemical names used: 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid 2,4-D), N-(phenylmethyl)- 1H-purin-6-amine (BA), N-(2-furanylmethyl)-1H-purin-6-amine (kinetin), alpha-cyclopropyl-alpha-(4-methoxy-phenyl)-5-pyrimidine methanol (ancymidol).
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