亚热带植物科学 ›› 2014, Vol. 43 ›› Issue (03): 212-215.DOI: 10.3969/j.issn.1009-7791.2014.03.006

• 植物生理与生化 • 上一篇    下一篇

大花蕙兰原生质体分离纯化

许传俊,陈益辉,李爱贞,黄珺梅,曾碧玉,黄雯   

  1. (1.福建省亚热带植物研究所,福建省亚热带植物生理生化重点实验室,福建 厦门 361006;2.集美大学 食品与生物工程学院,福建 厦门 361021)
  • 收稿日期:2014-05-13 修回日期:2014-05-13 出版日期:2014-09-10 发布日期:2014-09-10
  • 通讯作者: 许传俊
  • 作者简介:许传俊,博士,副研究员,从事植物细胞工程和细胞分子生物学研究。
  • 基金资助:

    福建省省属公益类研究院所基本科研专项(2009R10005-1)

Optimization Conditions of Cymbidium hybridum Protoplast Isolation

XU Chuan-jun,CHEN Yi-hui,LI Ai-zhen,HUANG Jun-mei,ZENG Bi-yu,HUANG Wen   

  1. (1.Fujian Key Laboratory of Subtropical Plant Physiology and Biochemistry, Fujian Institute of Subtropical Botany, Xiamen 361006, Fujian China; 2.College of Food and Biological Engineering, Jimei University, Xiamen 361021, Fujian China)
  • Received:2014-05-13 Revised:2014-05-13 Online:2014-09-10 Published:2014-09-10
  • Contact: XU Chuan-jun

摘要: 以大花蕙兰的叶片、根为试材,分析不同酶液组合、摇床转速、酶解温度、材料酶液比、酶解时间等因素对其原生质体分离的影响。大花蕙兰叶片、根经过90 min质壁分离预处理后,在温度28 ℃,于摇床上50 r·min-1振荡酶解6 h,叶、根分别在酶解液为纤维素酶2.0%+果胶酶0.1%+离析酶0.2%+崩溃酶0.2%和纤维素酶2.0%+果胶酶0.3%+离析酶0.1%+崩溃酶0.1%中得到最高产量的原生质体。

关键词: 大花蕙兰, 原生质体, 分离

Abstract: Taking Cymbidium hybridum leaves and roots as material, the effects of enzyme combination, shaking speed, enzymolysis time, enzymolysis temperature, and the ratio of materials enzyme on protoplast isolation were studied. The results showed that:after 90 min pretreatment for plasmolysis, protoplast yield from leaves was the highest using mixed enzyme (2.0% cellulose + 0.1% pectinase + 0.2% macerozyme + 0.2% driselase) with shaking speed 50 r·min-1 at 28 ℃ for 6 h. And protoplast yield from roots was the highest using mixed enzyme (2.0% cellulose + 0.3% pectinase + 0.1% macerozyme + 0.1% driselase) under the same conditions.

Key words: Cymbidium hybridum, protoplasts, isolation

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