浙江农业学报 ›› 2019, Vol. 31 ›› Issue (10): 1671-1680.DOI: 10.3969/j.issn.1004-1524.2019.10.13

• 植物保护 • 上一篇    下一篇

一株Fusarium oxysporum f. sp. niveum拮抗菌的筛选、鉴定及其抑菌特性

王恒煦1, 徐伟慧1,*, 杨友财2, 王志刚1, 刘泽平1, 王可昕1   

  1. 1.齐齐哈尔大学 生命科学与农林学院,黑龙江 齐齐哈尔 161006;
    2.黑龙江理工生物科技有限公司,黑龙江 哈尔滨 150000
  • 收稿日期:2019-03-04 出版日期:2019-10-25 发布日期:2019-10-30
  • 通讯作者: *,徐伟慧,E-mail: xwh800206@163.com
  • 作者简介:王恒煦(1995—),女,山东临沂人,硕士研究生,研究方向为土传病原菌与生防菌互作。E-mail: 1124081728@qq.com
  • 基金资助:
    国家自然科学基金青年科学基金(31700428); 黑龙江省自然科学基金联合引导项目(LH2019C068); 黑龙江省省属高等学校基本科研业务费科研项目(135209266)

Screening, identification, and characteristics of an antagonistic strain against Fusarium oxysporum f.sp. niveum

WANG Hengxu1, XU Weihui1,*, YANG Youcai2, WANG Zhigang1, LIU Zeping1, WANG Kexin1   

  1. 1. School of Life Science and Agriculture and Forestry, Qiqihar University, Qiqihar 161006, China;
    2. Heilongjiang Science-Engineering and Biotechnology Corporation, Harbin 150000, China
  • Received:2019-03-04 Online:2019-10-25 Published:2019-10-30

摘要: 为筛选西瓜枯萎病拮抗菌株并探究其抑菌特性,以齐齐哈尔市青昕蔬菜基地根际土为材料,采用平板对峙法筛选到1株能高效抑制西瓜专化型尖孢镰刀菌(Fusarium oxysporum f.sp. niveum,Fon)的拮抗菌株,通过形态特征、16S rDNA、gyrAgyrB序列分析鉴定其菌种;通过盆栽试验验证拮抗菌株对西瓜枯萎病的抑制作用;采用生长速率法研究拮抗菌株不同生长时期的无菌发酵滤液及其不同贮存条件下的抑菌能力;利用扫描电子显微镜(SEM)和共聚焦激光扫描显微镜(CLSM)观察并分析无菌发酵滤液对Fon孢子形态和膜完整性的影响。结果表明,拮抗菌株WD为Bacillus amyloliquefaciens,能有效抑制西瓜枯萎病的发生,抑制率达到57.57%,其无菌发酵滤液在衰亡期有较高的抑菌能力,对Fon菌丝生长抑制率为54.32%;无菌发酵滤液对温度、pH具有较高的耐受性,30 ℃、pH 7时抑菌活性最强,抑菌率可达58.36%;在4 ℃贮存至少45 d,且紫外光照120 min内对其抑菌活性没有影响;Fon孢子暴露在WD无菌发酵滤液下,孢子表面皱缩凹陷,膜完整性被破坏。综上所述,Bacillus amyloliquefaciens WD是1株高效的拮抗菌株,为西瓜枯萎病生防菌剂的研发与利用奠定了基础。

关键词: 西瓜专化型尖孢镰刀菌, 拮抗菌, 解淀粉芽孢杆菌, 无菌发酵滤液, 抑菌能力

Abstract: In order to screen an antagonistic strain against Fusarium oxysporum f.sp. niveum (Fon), the rhizosphere soil from vegetable crops on Qingxin farm in Qiqihar City was collected and used in this study. The antagonistic strain was screened out using plate confrontation method, and identified by morphological, 16S rDNA, gyrA and gyrB sequence analysis. Pot experiment was conducted to verify the control effect of antagonistic strains on Fusarium wilt of watermelon. Growth rate method was used to study antifungal activities of sterile fermentation filtrate from different growth stages of antagonistic strain and the stability of sterile fermentation filtrate. Fon conidia morphology and cell membrane integrity were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), respectively. The results showed that the antagonistic strain WD was Bacillus amyloliquefaciens, which could effectively inhibit the occurrence of watermelon wilt, and the inhibition rate reached 57.57%. The sterile fermentation filtrate had higher antifungal ability in the decline period, and the growth inhibition rate of Fon mycelium was 54.32%. Sterile fermentation filtrate from B. amyloliquefaciens WD had high tolerance to temperature and pH. The antifungal activity was the strongest at 30 °C and pH 7, the inhibition rate reached 58.36%; The sterile fermentation filtrate could store at 4 °C for at least 45 days, and UV light for 120 min had no effect on its antifungal activity. The Fon conidia were exposed to the sterile fermentation filtrate from B. amyloliquefaciens WD, the surface of the conidia collapsed and the membrane integrity was destroyed. In conclusion, B. amyloliquefaciens WD had high-efficiency antifungal activity on Fon, the results laid a foundation for development and utilization of biocontrol agent to Fusarium wilt of watermelon.

Key words: Fusarium oxysporum f.sp. niveum, Bacillus amyloliquefaciens, sterile fermentation filtrate, antifungal ability

中图分类号: