生物技术通报 ›› 2013, Vol. 0 ›› Issue (12): 178-183.

• 研究报告 • 上一篇    下一篇

亚硝化细菌原生质体化学诱变育种研究

董玉玮1, 2 张雁秋1 涂宝军1, 2 孙玲1, 2 曹文平1, 2   

  1. 1.中国矿业大学,徐州 221116
    2.徐州工程学院,徐州 221116
  • 收稿日期:2013-06-09 出版日期:2013-12-20 发布日期:2013-12-20
  • 作者简介:董玉玮,男,博士研究生,研究方向:环境微生物学;E-mail:dongyuwei66@163.com
  • 基金资助:
    江苏省自然科学基金项目(BK2011201),徐州工程学院科技计划项目(XKY2012216)

Chemical Mutagenesis Breeding of Protoplast of Ammonia-oxidizing Bacteria

Dong Yuwei1, 2, Zhang Yanqiu1, Tu Baojun1, 2, Sun Ling1, 2, Cao Wenping1, 2   

  1. 1. China University of Mining and Technology ,Xuzhou 221116
    2. Xuzhou Institute of Technology ,Xuzhou 221116
  • Received:2013-06-09 Published:2013-12-20 Online:2013-12-20

摘要: 以活性污泥中分离出的亚硝化细菌为研究对象,采用原生质体诱变技术,以化学诱变剂氯化锂、溴化乙锭诱变亚硝化细菌原生质体,选育亚硝酸盐氮富集能力高的菌株,同时研究了诱变剂浓度与脱氮能力之间的关系。结果表明,亚硝化细菌原生质体再生菌落的致死率随诱变剂浓度的增加而逐渐增大。采用1 L模拟污水培养基扩大培养后,经2.5 mg/mL氯化锂诱变的菌株LC002脱氮率为86.89%;经2 μg/mL溴化乙锭诱变的菌株EB003脱氮率为85.67%。两株诱变菌株亚硝态氮富集能力与未诱变菌株相比有明显的提高。原生质体诱变技术是一种选育优良亚硝化细菌的有效方式。

关键词: 亚硝化细菌, 原生质体, 诱变, 氯化锂, 溴化乙锭

Abstract: Strain with strong capacity of ammoxidation screened by repeated enrichment and separation methods from activated sludge, was collected from sewage treatment plant in China university of mining and technology. Lithium chloride and ethidium bromide mutagenesis methods of ammonia-oxidizing bacteria protoplast were used to breed mutant strain with strong capacity of ammoxidation. The relationship between concentrations of chemical mutagens and capacity of ammoxidation were researched. The results showed that regenerated colony number of protoplast decreased with increased lithium chloride or ethidium bromide mutagenic concentration. After culvation of 1 L simulation sewage medium, the mutagenic strain LC002, mutated by 2.5 mg/mL lithium chloride, still had better capacity of ammoxidation with 86.89% removal rate of ammonia nitrogen. The mutagenic strain EB003, mutated by 2 μg/mL ethidium bromide, also had better capacity of ammoxidation with 85.67% removal rate of ammonia nitrogen. Protoplast mutagenisis was an effective breeding method to breed excellent ammonia-oxidizing bacteria for further application.

Key words: Ammonia-oxidizing bacteria, Protoplast, Mutagenesis, Lithium chloride, Ethidium bromide