聚乙二醇诱导金钗石斛原生质体融合条件的初步探索

The priminary study of screening the optimum condition during the protoplast fusion in Dendrobium nobile with polyethylene glycol

目的在原生质体融合实验中筛选PEG分子量、工作浓度及孵育时间的最佳水平,为系统建立起金钗石斛原生质体融合技术体系奠定实验基础。方法研究采用药用品质优良的贵州赤水金钗石斛与茎粗生长快的云南瑞丽金钗石斛原生质体为材料,采取单因素分析法,研究不同的聚乙二醇(PEG)分子量、浓度及孵育时间对原生质体融合率的影响。结果 PEG的分子量对原生质体融合率有显著的影响。当分子量为3 350 Da,原生质体融合率最高,为18.32%;PEG浓度为40%时,原生质体融合率最高,为18.81%;孵育时间为25 min时,原生质体融合率最高,为18.62%。结论采用分子量为3 350Da的PEG在浓度为40%的条件下孵育25 min时,获得的原生质体融合率最高,为18.81%。

Objective To systemically construct the protocol of protoplast fusion in Dendrobium nobile Lindl,the optimum levels both of molecular weight and concentration of polyethylene glycol（ PEG） and of incubation time were screened during the fusion process. Methods In the current study,two D. Nobile landraces were applied as the material. One was originated from Chishui County of Guizhou Province with higher medical quality; another was from Ruili City of Yunnan Province with faster growing and larger stem diameter. The single factor analysis method was adopted as the experimental design and the protoplast fusion rate was selected as the criterion to screen the optimum levels of PEG molecular weight,PEG concentration and incubation time during the fusion experiment. Results The molecular weight of PEG significantly affected the protoplast fusion rate. The rate got the highest value as 18. 32%,18. 81% 18. 62%,when the molecular weight of PEG was 3 350 Da,PEG concentration was 40% and the incubation time was 25 min,individually. Conclusion The protoplast fusion rate could reach the highest value of 18. 81% under the conditions of the PEG molecular weight was 3 350 Da,the concentration was 40% and the incubation time was 25 min.