| 石斛兰叶片原生质体分离条件研究 |
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| 引用本文： | 罗丽华,严虹羽,王业华,沈通达.石斛兰叶片原生质体分离条件研究[J].安徽农业科学,2010,38(12):6144-6146. |
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| 作者姓名： | 罗丽华 严虹羽 王业华 沈通达 |
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| 作者单位： | 海南大学热带作物种质资源保护与开发利用教育部重点实验室,海南海口,570228;海南大学热带作物种质资源保护与开发利用教育部重点实验室,海南海口,570228;海南大学热带作物种质资源保护与开发利用教育部重点实验室,海南海口,570228;海南大学热带作物种质资源保护与开发利用教育部重点实验室,海南海口,570228 |
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| 摘 要： | 目的]探讨了石斛兰原生质体分离过程中的影响因素,建立最佳的原生质体制备体系。方法]以石斛兰叶片为材料,采用正交设计研究纤维素酶浓度、酶解时间、温度、pH值以及甘露醇浓度对石斛兰原生质体分离的影响。结果]以3%纤维素酶R-10＋0.6%果胶酶Y-23为混合酶液,0.5mol/L甘露醇为渗透压稳定剂,在26℃的黑暗条件下,pH值为5.4的酶液组合中,黑暗条件下振荡,酶解15h,获得的原生质体产量最高。结论]该制备体系为后续原生质体培养和体细胞杂交奠定良好的基础。
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| 关 键 词： | 石斛兰 原生质体 分离 |
Isolation Conditions of Protoplast in Dendrobium Leaves |
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| Institution: | LUO Li-hua et al(Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources(Hainan University),Ministry of Education,Haikou,Hainan 570228) |
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| Abstract: | Objective] To investigate the isolation conditions of protoplast in Dendrobium and establish a best protoplast preparation system.Method] Dendrobium leaves were used as research materials.The effects of cellulase concentration,hydrolysis time,temperature,pH value,and mannitol concentration on the separation of the protoplasts in Dendrobium were studied using orthogonal design.Result] The highest yield of protoplast was obtained in the conditions including the mixed enzyme solution of 3.0% Onozuka R-10+0.6% pectinase Y-23,the osmotic stabilizer of 0.50 mol/L mannitol,in dark and pH 5.4 for 15 h at 26 ℃.Conclusion] The preparation system lays a good foundation for subsequent protoplast culture and somatic cell hybridization. |
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| Keywords: | Dendrobium Protoplast Isolation |
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