A rapid and efficient alternative procedure for the regeneration of plants from hypocotyl protoplasts of Brassica napus

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Abstract
Protoplasts of several spring and winter varieties of Brassica napus were isolated from hypocotyl tissue. Protoplasts divided and formed cell colonies at high frequency, without browning when cultured in modified Shepards' medium. This high efficiency of proliferation was sustained through to plant regeneration with all varieties cultured. This has been attributed to the incorporation of a reservoir medium, the presence of 2,4-D in the proliferation medium, and the presence of kinetin in conjunction with lowering of the sucrose concentration in the regeneration medium.