生物技术通报 ›› 2023, Vol. 39 ›› Issue (12): 229-236.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0152

• 研究报告 • 上一篇    下一篇

贝莱斯芽孢杆菌ZHX-7的分离鉴定及抑菌促生效果

李莹(), 宋新颖, 何康, 郭志青, 于静, 张霞()   

  1. 山东省花生研究所,青岛 266100
  • 收稿日期:2023-02-22 出版日期:2023-12-26 发布日期:2024-01-11
  • 通讯作者: 张霞,女,博士,副研究员,研究方向:花生病害的综合防治;E-mail: zhangxia2259@126.com
  • 作者简介:李莹,女,博士,助理研究员,研究方向:花生病害的综合防治;E-mail: li1989ying0921@163.com
  • 基金资助:
    国家自然科学基金项目(31901940);国家自然科学基金项目(32202326);国家自然科学基金项目(31972967);山东省自然科学基金项目(ZR2021QD153);山东省自然科学基金项目(ZR2020QC131);青岛市自然科学基金项目(23-2-1-46-zyyd-jch);山东省农业科学院农业科技创新工程(CXGC2021B33);山东省农业科学院农业科技创新工程(CXGC2023A46)

Isolation and Identification of Bacillus velezensis ZHX-7 and Its Antibacterial and Growth-promoting Effects

LI Ying(), SONG Xin-ying, HE Kang, GUO Zhi-qing, YU Jing, ZHANG Xia()   

  1. Shandong Peanut Research Institute, Qingdao 266100
  • Received:2023-02-22 Published:2023-12-26 Online:2024-01-11

摘要:

为获得高效拮抗花生病害的生防微生物,以花生冠腐病菌为指示菌,采用稀释涂布平板法和平板对峙法,从山东莱西采集的花生连作田的病、健株根际土壤样品中筛选到1株高效生防菌株,编号为ZHX-7。结合形态学特征和16S rDNA、gyrB序列分析,确定ZHX-7为贝莱斯芽孢杆菌(Bacillus velezensis)。ZHX-7的菌悬液、无菌发酵液和挥发性气体对花生白绢病菌、基腐病菌、网斑病菌和轮斑病菌菌丝生长的抑制均达到极显著的水平,平板对峙试验表明ZHX-7的菌悬液对花生白绢病菌、基腐病菌、网斑病菌和轮斑病菌的抑菌率分别为64.40%、70.37%、77.43%和65.32%。在以ZHX-7无菌发酵液稀释10倍的PDA培养基上,花生白绢病菌、基腐病菌、网斑病菌和轮斑病菌菌丝的生长抑制率分别为67.81%、52.16%、23.52%和69.93%。双皿对扣试验表明ZHX-7的挥发性气体对花生白绢病菌、基腐病菌、网斑病菌和轮斑病菌的抑菌率分别为86.82%、35.40%、81.70%和44.19%。ZHX-7可产生蛋白酶、纤维素酶和淀粉酶等次级代谢产物;此外,在温室盆栽条件下,ZHX-7有菌发酵液明显促进了花生的生长,同对照相比,鲜质量、干质量分别增加31.24%和26.47%,并且ZHX-7有菌发酵液明显提高了花生对白绢病的抗性,防效达到51.76%。综上所述,本研究筛选获得的一株贝莱斯芽孢杆菌ZHX-7,能够抑制多种花生病原真菌,同时促进花生生长,是一株具有宽广拮抗谱的多功能菌株,具有较好的生防应用潜力。

关键词: 贝莱斯芽孢杆菌, 花生病害, 分离鉴定, 抑菌率, 生物防治, 促生作用, 白绢病菌

Abstract:

In order to obtain highly effective biocontrol microorganism for controlling peanut diseases, using Aspergillus niger as the indicator, a highly effective biocontrol strain ZHX-7 was screened from soil samples by agar disk dilution and dual-culture assay. The soil samples were collected from the rhizosphere of diseased and healthy peanut plants in continuous cropping field in Laixi, Shandong province. The strain ZHX-7 was identified as Bacillus velezensis based on the morphological characteristics, 16S rDNA and gyrB sequence analysis. The bacterial suspension, sterile fermentation broth and volatile gas of ZHX-7 significantly inhibited the growth of Sclerotium rolfsii, Fusarium neocosmosporiellum, Phoma arachidicola and Alternaria alternata. The antagonistic experiment results showed that the inhibition rates of ZHX-7 suspension against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 64.40%, 70.37%, 77.43% and 65.32%, respectively. In the sterile fermentation broth of ZHX-7 diluted 10 times with PDA medium, the inhibition rates of ZHX-7 against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 67.81%, 52.16%, 23.52% and 69.93%, respectively. The double dish test showed that the inhibition rates of ZHX-7 volatile gas against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 86.82%, 35.40%, 81.70% and 44.19%, respectively. ZHX-7 produced secondary metabolites such as protease, cellulose and amylase. In addition, under the condition of pot culture in greenhouse, ZHX-7 fermentation broth significantly promoted the growth of peanut. Compared with the control, the fresh mass and dry mass increased by 31.24% and 26.47%, respectively. ZHX-7 fermentation broth improved the resistance of peanut to S. rolfsii, and the control effect reached 51.76%. In conclusion, a strain of B. velezensis ZHX-7 screened in this study may inhibit a variety of peanut pathogenic fungi and promote peanut growth at the same time. It is a multifunctional strain with broad antagonistic spectrum against peanut diseases, and high potential for biocontrol application.

Key words: Bacillus velezensis, peanut diseases, isolation and identification, antibacterial rate, biological control, growth-promoting effect, Sclerotium rolfsii