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叶用芥菜品种‘冲辣菜’(Brassica juncea var.longeptiolatus cv,‘Chonglacai’)试管苗叶片原生质体用液体浅层培养。培养后48—72h观察到第一次分裂。培养7—10天后每隔一周添加甘露醇浓度逐渐降低(6％—4％—0％)的新鲜培养基,一共加3次。培养过程中不时地慢速振荡有利于细胞的持续分裂,从而促进细胞团和微愈伤组织的形成。6周后形成直径达0.5—1.0mm的微愈伤组织。植板率(微愈伤组织数占培养原生质体数的百分率)为0.2％。微愈伤组织转至附加萘乙酸(NAA)0.2、6-苄氨基嘌呤(BAP)2.0mg/L 的MS固体培养基上,10天后分化出芽。当这些芽长成 2—3 cm高的新梢后转至含NAA 1.0mg/L和赤霉素(GA_3)0.2mg/L或只附加吲哚乙酸(IAA)1.0mg/L的MS培养基上分化出根,形成了完整的小植株。
Leaf protoplasts from leaves of Brassica juncea var.longeptiolatus cv (’Chonglacai’), a mustard variety, were cultured in shallow liquid. The first division was observed 48-72 h after culture. After 7-10 days of culture, fresh medium with a gradually decreasing mannitol concentration (6% -4% -0%) was added every other week for a total of 3 times. Slow oscillations from time to time during culture favor sustained division of the cells, thereby promoting the formation of cell clumps and micro-callus. Micro-callus, 0.5-1.0 mm in diameter, formed after 6 weeks. The plating rate (the percentage of micro-callus to the number of cultured protoplasts) was 0.2%. Micro-callus was transferred to MS solid medium supplemented with NAA 0.2 mg / L 0.2 mg / L NAA, and budding occurred after 10 days. When these shoots grew 2-3 cm high shoots, they were transferred to MS medium containing 1.0 mg / L NAA and 0.2 mg / L gibberellic acid (GA 3) or only 1.0 mg / L indole acetic acid (IAA) On the differentiation of the root, forming a complete plantlets.