Induction of Hormone Autonomous Suspension Culture derived from Leaf Explants of Sugar Beets

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Hormone autonomous callus was initiated reproducibly on MS medium with 0.25mg/l BA as a sole plant hormone (AI medium) from 4-week-old leaf explants of sugar beet, whereas 2,4-D requiring callus were initiated on AI medium to which 0.1mg/l 2,4-D was added. On the other hand, when leaf explants were inoculated into liquid AI medium and cultured on a reciprocal shaker, single cells began to be released from the edge of explants at 6 days after inoculation of explants, followed by active release. When single cells obtained were transferred to fresh liquid MS medium without plant hormone, they could olivide and grow autonomously, giving rise to hormone autonomous suspension cultures. Whereas single cells released in MS medium without BA seldom formed cell cluster in MS medium with or without BA. Accordingly only single cells released from the leaf explants treated with BA could form cell clusters, but in this case, the formation of clusters from single cells proceeded regardless of the presence and alsence of BA. On the other hand, when single cells were cultured in the conditioning medium of hormone autonomous suspension cultured cells, cell clusters, differed from autonomous cell clusters were formed and adventive root from them was observed to be formed.