石斛兰叶片原生质体分离条件研究  

Isolation Conditions of Protoplast in Dendrobium Leaves

作　　者：罗丽华[1] 严虹羽[1] 王业华[1] 沈通达[1] 

机构地区：[1]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

出　　处：《安徽农业科学》2010年第12期6144-6146,共3页Journal of Anhui Agricultural Sciences

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。[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.

关 键 词：石斛兰 原生质体 分离 

分 类 号：S682.31[农业科学—观赏园艺]