湖北农业科学 ›› 2022, Vol. 61 ›› Issue (18): 78-83.doi: 10.14088/j.cnki.issn0439-8114.2022.18.012

• 植物保护 • 上一篇    下一篇

微生物源生防杀菌剂的筛选方法研究

张志刚, 王开梅, 吴兆圆, 万中义, 方伟   

  1. 湖北省生物农药工程研究中心/湖北省农业科技创新中心生物农药分中心,武汉 430064
  • 收稿日期:2022-07-01 出版日期:2022-09-25 发布日期:2022-10-21
  • 通讯作者: 王开梅(1969-),男,湖北武汉人,研究员,硕士,主要从事微生物农药研究,(电话)18627065137(电子信箱)wang_kaimei@163.com。
  • 作者简介:张志刚(1971-),男,湖北天门人,副研究员,主要从事微生物农药研究,(电话)13638694161(电子信箱)yilu98@163.com。
  • 基金资助:
    国家重点研发计划重点专项(2017YFD0201301-4); 湖北省农业科技创新中心创新团队项目(2019-620-000-001-27)

Study on screening methods of bio-fungicide from microorganism

ZHANG Zhi-gang, WANG Kai-mei, WU Zhao-yuan, WAN Zhong-yi, FANG Wei   

  1. Hubei Biopesticide Engineering Research Center/The Branch Center of Biopesticide, Hubei Agricultural Scientific and Technological Innovation Center, Wuhan 430064,China
  • Received:2022-07-01 Online:2022-09-25 Published:2022-10-21

摘要: 以胶孢炭疽菌(Colletotrichum gloeobosporioides)、番茄枯萎病菌(Fusarium oxysporum)、灰霉病菌(Botrytis cinerea)、西瓜猝倒病菌(Pythium aphanidematum)作为模式靶标,使用半固体培养基法(PDB培养基加入适量琼脂)及平板对峙法(PDA培养基),对真菌及放线菌杀菌剂标准菌株的发酵液、上清液和菌丝进行了杀菌活性比较。结果表明,样品的活性与筛选方法和培养时间有直接关联。半固体培养基法中发酵液及菌丝的活性分辨度差,而对真菌及放线菌标准菌株的上清液的活性反应清晰;平板对峙法中,随着培养时间的增加,发酵液及菌丝的活性增强,而真菌及放线菌杀菌剂标准菌株的上清液的活性减弱。半固体培养基法适用于针对不含活菌的上清液的筛选,平板对峙法适用于针对含有活菌的发酵液、菌丝和上清液的筛选,根据样品的类型选择适当的筛选方法和培养时间可以减少假阴性结果。

关键词: 模式靶标, 生物杀菌剂, 半固体培养基法, 平板对峙法

Abstract: Using Colletotrichum gloeobosporioides, Fusarium oxysporum, Botrytis cinerea, Pythium aphanidematum as the model targets, six fungi standard strains and six actino standard strains were tested by semi-solid medium method (PDB medium with the appropriate amount of agar) and disk diffusion method (PDA medium). The activities of fermentation broth, supernatant and mycelium of fungi and actino were compared by semi-solid medium method and agar disk diffusion method. The results showed that the activity of the sample was directly related to the screening method and incubation time. The activity resolution of fermentation broth and mycelium in semi-solid medium method was blurring, and the activity response to the supernatant of standard strains of fungi and actino was clear; in the agar disk diffusion method, with the increase of culture time, the activity of fermentation broth and mycelium increased, while the activity of the supernatant of standard strains of fungi and actinomycetes fungicides decreased. The semi-solid medium method was suitable for the screening of supernatants without living cells, and the agar disk diffusion method was suitable for the screening of fermentation broth and mycelium with living cells. Selecting the appropriate screening method and culture time according to the type of sample could reduce the false negative results.

Key words: mode target, bio-fungicide, semi-solid medium method, disk diffusion method

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