生物技术通报 ›› 2013, Vol. 0 ›› Issue (4): 96-100.

• 研究报告 • 上一篇    下一篇

尖孢镰刀菌辣椒专化型原生质体制备条件优化

弭宝彬1, 张吉祥2, 杨宇红2, 茆振川2   

  1. 1. 中南大学研究生院隆平分院,长沙 410125 ;2. 中国农业科学院蔬菜花卉研究所,北京 100081
  • 收稿日期:2012-10-24 修回日期:2013-04-22 出版日期:2013-04-22 发布日期:2013-04-22
  • 作者简介:弭宝彬,男,硕士研究生,研究方向:蔬菜学;E-mail:mibaobin@126.com
  • 基金资助:
    农业公益性行业科研专项(200903049)

Optimizing Method of Preparation of Protoplast in Fusedum oxysporum f. sp. capsicum

Mi Baobin1, Zhang Jixiang2, Yang Yuhong2, Mao Zhenchuan2   

  1. 1. Longping Branch of Graduate School,Central South University,Changsha 410125 ;2. Institute of Vegetable and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081
  • Received:2012-10-24 Revised:2013-04-22 Published:2013-04-22 Online:2013-04-22

摘要: 以尖孢镰刀菌辣椒专化型(Fusedum oxysporum f. sp. capsicum)为供试菌株,研究了其原生质体制备的最佳条件, 明确了菌龄、酶的种类、酶解时间、酶解温度及转速等因素对原生质体制备的影响。结果表明,当利用孢子培养的14 h 的菌丝体 以15 mg/mL 崩溃酶、0.7 mol/L 的NaCl 作为渗透压稳定剂,140 r/min,32℃酶解4 h时,可最大限度的收集原生质体,其在再生固 体培养基SR 上再生率为32.3%。

关键词: 尖孢镰刀菌辣椒专化型, 原生质体制备, 再生

Abstract: Several conditions in preparing the protoplast of Fusedum oxysporum f. sp. capsicum were optimized. Effects of the incubation time, enzyme system, the enzymolysis time, temperature and rotate speed were discussed. It was found that when keeping other parameters as : spore cultured for 14 hours, 15 mg/mL of Drislase, 0.7 mol/L NaCl and 140 r/min, 32℃ for 4 h, the minimum consummation of protoplasts can be obtained and the regeneration rate of the protoplast was 32.3% on regeneration medium SR.

Key words: Fusedum oxysporum f. sp. capsicum, Preparation of protoplast, Regeneration