Study on screening, identification, culture optimization, and biocontrol effects of antagonistic bacteria against tobacco leaf mildew

doi:10.3969/j.issn.1004-1524.20240815

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (7): 1481-1491.DOI: 10.3969/j.issn.1004-1524.20240815

• Plant Protection • Previous Articles Next Articles

Study on screening, identification, culture optimization, and biocontrol effects of antagonistic bacteria against tobacco leaf mildew

ZHOU Hang¹(), JIA Tao², FANG Jiangping³, YANG Yongfeng⁴, YU Yangyang¹, QIU Yao¹, CHEN Siyuan¹, FENG Wei¹, ZHANG Jingyuan¹, CHEN Hongli¹^,^*()

1. Tobacco College, Henan Agricultural University, Zhengzhou 450046, China
2. Tianchang International Tobacco Co., Ltd., Xuchang 461000, Henan, China
3. China Tobacco Jiangsu Industrial Co., Ltd., Nanjing 210015, China
4. Technology Center, China Tobacco Henan Industrial Co., Ltd., Zhengzhou 450016, China

Received:2024-09-19 Online:2025-07-25 Published:2025-08-20

Abstract

Abstract:

To identify the pathogenic fungi responsible for moldy stored tobacco leaves in Yunnan tobacco-growing areas and screen antagonistic strains against these pathogens, this study isolated fungi from moldy tobacco leaves. Pathogens were identified through Koch’s postulates verification, cultural characteristics, and molecular biology. Antagonistic strains were screened from healthy tobacco leaves using the plate dilution method, followed by physiological-biochemical characterization and culture condition optimization. The biocontrol efficacy of antagonistic strains against tobacco mold at varying concentrations was validated. The results demonstrated that two pathogenic strains isolated from moldy tobacco leaves in Yunnan were identified as Aspergillus niger and Aspergillus flavus based on morphological and molecular characteristics. An antagonistic strain, Bacillus velezensis ZH-X10, was screened and found to induce mycelial deformation and shrinkage in pathogens, as observed via scanning electron microscopy (SEM). Optimal fermentation conditions for ZH-X10 were pH 7.0, 35 ℃, and NaCl mass fraction of 2%. A concentration of 10⁸ CFU·mL^-1 of ZH-X10 demonstrated effective control against A. niger and A. flavus, representing the threshold inoculation concentration for mold suppression. In conclusion, Bacillus velezensis ZH-X10 exhibited significant antagonistic activity against tobacco mold pathogens and showed potential as a novel biocontrol agent.

Key words: tobacco leaf, storage, fungus, mold, antagonistic bacteria, biological control

CLC Number:

S432.44

ZHOU Hang, JIA Tao, FANG Jiangping, YANG Yongfeng, YU Yangyang, QIU Yao, CHEN Siyuan, FENG Wei, ZHANG Jingyuan, CHEN Hongli. Study on screening, identification, culture optimization, and biocontrol effects of antagonistic bacteria against tobacco leaf mildew[J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1481-1491.

Figures/Tables 12

Fig.1 Cultivation characteristics of ZZ-1 and ZZ-2 colonies

Fig.2 Phylogenetic trees of strains ZZ-1 and ZZ-2

Fig.3 Infection symptoms of tobacco leaves inoculated with Aspergillus niger and Aspergillus flavus CK, Inoculated with sterile water; A, Inoculated with Aspergillus niger; B, Inoculated with Aspergillus flavus.

Fig.4 Initial screening effect of antagonistic strains A, Aspergillus niger; B, Aspergillus flavus.

Fig.5 Rescreening results of antagonistic strain ZH-X10 A, Aspergillus niger control; B, Aspergillus flavus control; C, Aspergillus niger re-screening; D, Aspergillus flavus re-screening.

Fig.6 Effect of antagonistic strain ZH-X10 on hyphae growth of Aspergillus niger and Aspergillus flavus A, Aspergillus niger control; B, Aspergillus niger treated with antagonistic strain ZH-X10; C, Aspergillus flavus control; D, Aspergillus flavus treated with antagonistic strain ZH-X10.

Fig.7 Phylogenetic tree of strain ZH-X10 based on 16S rRNA

Fig.8 Growth curve of the strain ZH-X10

Fig.9 Effects of pH value on the growth of strain ZH-X10

Fig.10 Effect of temperature on the growth of strain ZH-X10

Fig.11 Effect of NaCl mass fraction on the growth of strain ZH-X10

Fig.12 Control effect of ZH-X10 suspension against mold in tobacco leaves CK, Control group, inoculated with sterile water; T1-T4 were inoculated with ZH-X10 bacterial suspension of 106,107,108,109 CFU·mL-1, respectively.

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Study on screening, identification, culture optimization, and biocontrol effects of antagonistic bacteria against tobacco leaf mildew

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