湖北农业科学 ›› 2020, Vol. 59 ›› Issue (10): 77-80.doi: 10.14088/j.cnki.issn0439-8114.2020.10.015

• 植物保护 • 上一篇    下一篇

贝莱斯芽孢杆菌CY30菌株的常压室温等离子体诱变育种研究

刘芳, 廖先清, 周荣华, 饶犇, 陈伟, 黄大野, 万中义   

  1. 湖北省生物农药工程研究中心,武汉 430064
  • 收稿日期:2020-02-18 出版日期:2020-05-25 发布日期:2020-08-03
  • 通讯作者: 黄大野,副研究员,博士,主要从事生物杀菌剂方面的研究,(电子信箱)xiaohuangdaye@126.com;万中义,研究员,博士,主要从事微生物天然产物方面的研究,(电子信箱)wanzhongyi1964@126.com。
  • 作者简介:刘 芳(1978-),女,湖北天门人,助理研究员,主要从事微生物农药发酵工艺研究,(电话)18086025680(电子信箱)24807004@qq.com。
  • 基金资助:
    国家重点研发计划项目(2017YFD0201107);湖北省技术创新专项(2017ABA160);湖北省农业科技创新中心创新团队项目(2019-620-000-001-27)

Studies on mutagenesis of Bacillus velezensis CY30 with atmospheric and room temperature plasma

LIU Fang, LIAO Xian-qing, ZHOU Rong-hua, RAO Ben, CHEN Wei, HUANG Da-ye, WAN Zhong-yi   

  1. Hubei Biopesticide Engineering Research Centre,Wuhan 430064,China
  • Received:2020-02-18 Online:2020-05-25 Published:2020-08-03

摘要: 采用常压室温等离子体诱变贝莱斯芽孢杆菌(Bacillus velezensis)CY30菌株。菌株的致死率强度结果表明,在电源功率为100 W,处理距离2 mm,工作气流量10 L/min时,等离子体对CY30菌株诱变最优处理时间为50 s。通过显微镜检初筛诱变菌株,并利用摇瓶发酵结合芽孢数测定和杀菌活性评价进行复筛,建立完整的诱变筛选体系。通过斜面镜检初筛、摇瓶三轮复筛,获得摇瓶液芽孢数122.3亿/mL,杀菌活性75.58%,且遗传稳定的突变菌株CY30-314。突变株发酵水平较出发菌株芽孢数提高25.3%,杀菌活性提高15.6%,发酵周期缩短了2 h。

关键词: 常压室温等离子(ARTP), 贝莱斯芽孢杆菌(Bacillus velezensis), 芽孢数, 杀菌活性

Abstract: Atmospheric and room temperature plasma(ARTP) was used to mutate Bacillus velezensis CY30 for improving the production of spores and fungicidal activity. The most optimal condition for the ARTP mutagenesis was working flow rate at 10 L/min, treating distance of 2 mm and electrical power of 100 W. The mutants were selected based on microscopic morphology of the colonies, the spore counts and antifungal activity of the cultures from shake-flask fermentation. A mutant with code of CY30-314 was obtained through 3 rounds of microscopic checking of the slants and shake-flask fermentation. The spore counts of the mutant in shake-flask fermentation was 12.23 billion spores/mL, and the corresponding cultures inhibited the growth of fungus for 75.58%. The mutant also showed genetic stability. The mutant produced 25.3% more spores in shake-flask fermentation which resulted in the increase of antifungal activity at 15.6%. The duration of the mutant was shorted for 2 h.

Key words: Atmospheric and room temperature plasma(ARTP), Bacillus velezensis, spore count, antifungal activity

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