Shoot regeneration from protoplasts of Ulmus × ‘Pioneer’
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Protoplasts were isolated from leaf-derived calli of Ulmus × 'Pioneer' with a yield of 0.2 × 103-2.0 × 105 protoplasts/gram of callus. Protoplasts were cultured in KM medium 8p supplemented with 1.0 μM 2,4-Dichlorophenoxyacetic acid (2,4-D), 2.5 μM naphthaleneacetic acid (NAA), 1 μM zeatin, 0.38 M glucose and 0.7 mM sucrose. Cultures were incubated in darkness at 25°C. Cell wall regeneration followed by cell division was observed in at most 20% of the protoplasts 10 days after culture. However, colonies (0.2-0.3 mm in diameter) developed from less than 7% of the total cells 14 days after culture. A dense green callus formed 6 weeks after colonies were transferred to Murashige and Skoog (MS) medium containing 200 mg/l casein hydrolysate, 3% (w/v) sucrose, and different concentrations of benzyladenine (BA) or kinetin (5, 10, 20, 30 μM). Shoot bud primordia were initiated on MS medium containing 10-30 μM BA. Buds developed into shoots three weeks after they were transferred to MS medium containing 2 μM BA. Shoots were rooted and adapted to the green-house environment. © 1986.
author list (cited authors)
Sticklen, M. B., Domir, S. C., & Lineberger, R. D.