When the callus produced globular-stage embryos on MS solid
medium with 2,4-D and 3% sucrose, the globular embryos were
removed and transferred to 500 mL-Erlenmeyer flasks containing
200 mL of liquid MS medium supplemented with 2,4-D and 3%
sucrose for further growth. The liquid cultures were agitated at
100 rpm on a gyratory shaker in the dark. After 1 mo of culture, the
proliferated globular embryos in flasks were transferred to indi-
vidual petri dishes containing solid MS medium with gibberellic
acid (GA 3 ) and 3% sucrose for maturation and germination of
embryos.