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用酶法从结球甘蓝(Brassica oleracea L.var.capitata)F_1代杂种‘报春’的第一真叶分离原生质体,培养在修改的DPD液体培养基(CaCl_(2)·2H_(2)O800毫克/升,补加2,4-D0.5毫克/升、激动素KT1毫克/升、甘露醇0.3M、蔗糖20克/升)中,获得了持续的细胞分裂和体细胞胚胎发生。细胞团转至修改的MS固体培养基上诱导出愈伤组织。愈伤组织转至MS加KT 3毫克/升、赤霉酸GA_(3)0.1毫克/升的分化培养基上后,分化出了大量丛生芽。分化的芽在MS加吲哚乙酸(IAA)、KT、 GA_(3)各0.1毫克/升、并补加N.Z.amine 500毫克/升的培养基上,形成了植株。
Protoplasts were isolated from the first true leaves of Brassica oleracea L.var.capitata F_1 hybrid ’Primula’ by enzymatic method and cultured on modified DPD liquid medium (CaCl 2 2H 2) O800 mg / L, supplemented with 2,4-D0.5 mg / L, KT1 mg / L kinetin, 0.3 M mannitol, 20 g / L sucrose), sustained cell division and somatic embryogenesis were obtained. Transfer the cell pellet to a modified MS medium to induce callus. Callus was transferred to MS plus KT 3 mg / L, gibberellic acid GA_ (3) 0.1 mg / L differentiation medium, the differentiation of a large number of clusters of buds. The differentiated shoots formed on the medium of MS plus indole acetic acid (IAA), KT, GA_ (3) each 0.1 mg / L supplemented with N.Z.amine 500 mg / l to form the plant.