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作 者 ：李 贵1,2,李必元2,王五宏2,岳智臣2,钟新民2*,侯喜林1

期 刊 ：西北植物学报 2012年 12期 页码：2438~2443

关键词：结球甘蓝；下胚轴；原生质体培养；植株再生；

Keywords:cabbage (Brassica oleracea L.var capitata), hypocotyl, protoplast culture, plant regeneration,

摘 要 ：以结球甘蓝‘新夏50’的无菌苗下胚轴为材料，对影响原生质体分离、纯化与培养的主要因素进行研究，建立适合结球甘蓝原生质体游离、纯化、收集、培养以至再生出完整植株的实用技术体系，为其非对称细胞融合及品种改良与创新等研究奠定基础。结果表明：2.5%纤维素酶R-10+0.05%果胶酶Y-23+9CPW+5 mmol/L MES的混合酶液，从4 d苗龄的下胚轴上分离出高产率的原生质体。在改良B₅+0.5 mg/L 2,4-D+0.2 mg/L 6-BA+0.2 mg/L NAA的液体培养基上，原生质体分裂旺盛。形成愈伤组织后经芽诱导和生根培养，获得了再生植株。倍性检测结果表明，不同原生质体所获得的24株再生植株中，19株为正常二倍体，4株为嵌合体，1株为四倍体。

Abstract:The main factors affect cabbage hypocotyl protoplast isolation,purification and cultivation were studied,in order to establish a practical technology system of isolation,purification,collection,culture and eventually a complete plant regeneration for cabbage protoplast.Then establish a basis for latter research such as asymmetric cell fusion and finally provide the conditions for cabbage variety improvement and innovation.The results are as follows：Protoplasts were isolated by enzyme digestion from hypocotyl of 4 day-old seedling of head cabbage.The optimum enzyme combination was attained with 2.5% cellulase R-10+0.05% pectolase Y-23+9CPW+5 mmol/L MES.In the basic medium of improved B₅ supplemented with 0.5 mg/L 2,4-D,0.2 mg/L 6-BA,0.2 mg/L NAA,the cells divided luxuriantly.Regenerated plants were formed from callus after bud induction and root initiation.In 24 regenerated plants,19 were normal diploid,4 were diploid/tetraploid chimera and 1 was tetraploid.

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