Abstract
An extraction solution that is optimal for protoplast release in Eragrostis tef (Zucc.) Trotter and Sorghum bicolor (L.) Moench consists of 1% Cellulysin, 0.5 % Macerozyme and 2 % Cellulase Onozuka, 1 mM CaCl2, 5 mM 2, (n-morpholino) ethane sulfonic acid, and 0.4 M mannitol. A culture medium consisting of two parts of Gamborg et al. medium, three parts of Murashige and Skoog medium, 1 % Schenk and Hildebrandt powder vitamin, 1 mg/l matin, 10 mM glutamine, 10 mg/l of 2,4-dichlorophenoxyacetic acid, and 10 mg/l naphthaleneacetic acid, was optimal for the survival and division of a small fraction of cultured protoplasts and protoplast fusion products. Electroporation and heat shock treatments stimulated cell wall formation and cell division in both sorghum and tef. Electrical parameters and conditions for fusing tef and sorgum were determined. Field pulse conditions for eventual electrogene transfer in tef and sorghum consisted of 700 and 400 V/cm, respectively, at 20°C and 40μs field pulse duration.
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