安徽农业科学2012，Vol.40Issue(3)：1374-1376,3.
金耳原生质体制备与再生研究
Research on Protoplast Preparation and Regeneration Conditions of Tremella aurantialba
贺婷 1郭成金1
作者信息
- 1. 天津师范大学生命科学学院,天津300387
- 折叠
摘要
Abstract
[Objective] This study aimed to explore the protoplast preparation and regeneration conditions of Tremella aurantialba. [Method] Optimal combination of five factors affecting the protoplast preparation and regeneration of Tremella aurantialba was selected by using orthogonal experiment, including enzyme system, culture age, enzymolysis temperature, enzymolysis duration and osmotic stabilizer. [Result] The results showed that there were three release modes of Tremella aurantialba protoplasts: release from top in the early period of enzymolysis, release from the side and release in situ. The optimal protoplast preparation condition was selecting mycelium cultured in liquid medium for 3 d, for enzymolysis in mixed enzyme solution containing 1% of cellulase + 1% of snailase + 1% of lywallzyme at 36 ℃for 4 h, with 0.6 mol/L sucrose as osmotic stabilizer, and the obtained preparation rate had reached 1.76 x 107 protoplasts/ ml. The optimal regeneration condition was using mycelium cultured in liquid medium for 5 d, for enzymolysis in enzyme solution containing 1.5% of lywallzyme at 33 ℃for 3 h, with 0. 6 mol / L sucrose as osmotic stabilizer, and the regeneration rate had reached 0. 22% . [Conclusion] This study provided valuable support for protoplast fusion, ultraviolet mutation breeding and genetic engineering research.关键词
金耳/原生质体/制备/再生/正交设计Key words
Tremella aurantialba/ protoplast/ preparation/ regeneration/ orthogonal experiment design分类
农业科技引用本文复制引用
贺婷,郭成金..金耳原生质体制备与再生研究[J].安徽农业科学,2012,40(3):1374-1376,3.
