Callus formation from protoplasts derived from cell suspension cultures of Vicia faba L. [broad beans]
1981
Roeper, W. (Hamburg Univ. (Germany, F.R.). Inst. fuer Allgemeine Botanik)
High yields of protoplasts were obtained by enzymatic treatment of an established cell suspension culture of Vicia faba L. The release of protoplasts was achieved in an enzyme solution containing 4 percent cellulase Onozuka R-10, 0,5 percent Pectinase Sigma and 0,08 percent Rohament P, salts, mannitol and sorbitol 0,45 osmolal within 6 hours at 25 degrees Celsius. The protoplasts were washed by sedimentation in 60 percent seawater. The modified culture medium (Kao and Michayluk, 1975) contained 0,5 mg/l indoleacetic acid, 0,2 mg/l 2,4-dichlorphenoxyacetic acid amd 0,5 mg/l 6-benzylaminopurine. The first cell divisions were observed 3 or 4 days after isolation. When transferred to semi solid media the protoplasts formed cell colonies that gave rise to proliferating calli
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