霍山石斛原生质体制备条件初探

Study on Protoplast Preparation of Dendrobioum huoshanense

本实验采用机械法和酶解法对霍山石斛原生质体的分离条件进行研究。结果表明,机械法制备霍山石斛原生质体,得率很低。正交试验法优选出的酶解法最佳组合条件为:纤维素酶1.5%+果胶酶1.5%+离析酶1.0%的酶液组合(溶于0.6mol/L的甘露醇),pH值5.4,28℃温育18 h。

Mechanical method and enzymic method are employed for the preparation of protoplasts of Dendrobium huoshanense in this study. The results show that few protoplasts may be gained from the leaf cells by mechanical method. The results of orthogohal test for protoplasts preparation by enzymic method suggest the best combination is as follows：Onozuka R-10 1.5%＋ Pectolase 1. 5% ＋ Macerozyme R-10 1.0% （ dissolved in 0. 6 mol/L mannitol）, p H 5. 8,28℃, incubating for 18 hours.