结球甘蓝下胚轴原生质体培养再生植株体系的优化研究
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浙江省重大科技专项(2009C2006-1);浙江省蔬菜产业科技创新团队项目(2009R50026)


Plant Regeneration from Hypocotyl Protoplast Culture of Cabbage (Brassica oleracea L.var capitata)
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    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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李 贵,李必元,王五宏,等.结球甘蓝下胚轴原生质体培养再生植株体系的优化研究[J].西北植物学报,2012,32(12):2438-2443

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  • 在线发布日期: 2013-01-06
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