Establishment of Embryogenic Soybean Suspension Cultures

Abstract

Immature cotyledons of soybean were placed on the medium containing MS (salts), B₅ (vitamins) and 10, 20, 30 and 40 mg/l 2, 4-D. High percentage of embryogenesis was observed on the medium which contained 20 mg/l 2, 4-D. The medium which contained 3% sucrose gave higher percentage of embryogenesis than the medium containing 6% sucrose. Embryos and embryogenic callus produced on the medium containing 20 and 30 mg/l 2, 4-D were placed into suspension culture medium containing MS (salts), B₅ (vitamins), 15 mM glutamine, 5 and 10 mg/l 2, 4-D and 6% sucrose. Embryogenic suspension cultures proliferated as clumps of globular embryos and were maintained by subculturing every four weeks for over 6 months. Embryo development was observed when larger clumps of globular embryos were transferred to the MM (Maturation Medium) medium for embryo maturation.