Isolation, identification and characterization of an antagonistic bacterium against Bipolaris papendorfii

ZHANG Xin-Jie1 LIU Yan-Ce1 QIAN Xin-Yu1 XIAO Sheng-Lin1 YU Bo1 DONG Jin-Gao2 HAO Zhi-Min1

(1.College of Life Sciences, Hebei Agricultural University, Baoding, Hebei, China 071001)
(2.College of Plant Protection, Hebei Agricultural University, Baoding, Hebei, China 071001)

【Abstract】 [Background] Bipolaris papendorfii is the main fungal pathogen of Curvularia leaf spot, and harmful to agricultural production. At present, the use of bacteria to control B. papendorfii becomes a research focus in this field. [Objective] To screen the strains with high antagonistic activity against B. papendorfii, and identify the antagonistic mechanism. [Methods] Twenty-two bacterial strains were isolated from the soil under the surface of maize field by the method of plate confrontation. One strain L-14 with high antagonistic activity to B. papendorfii was obtained by repeated screening. [Results] Strain L-14 with the highest antagonism was obtained by using the plate diffusion method. After morphological observation, 16S rRNA gene sequence analysis, physiological and biochemical characteristics detection, the strain was finally identified as Bacillus subtilis. Moreover, the strain is a wide-spectrum antagonistic biocontrol strain with inhibitory effects on Fusarium proliferatum, Fusarium graminearum, Exserohilum turcicum, Bipolaris papendorfii and Botrytis cinerea. Then, we studied the antibacterial mechanism of strain L-14 and obtained that the fermentation crude protein extracts had no effect on the morphology of the aerial hyphae of B. papendorfii but can cause substrate mycelial aberration, and had an inhibitory effect on spore germination. [Conclusion] The antagonistic bacterium B. subtilis obtained in this study has a broad spectrum and high antagonistic activity in the control of plant diseases, and can effectively control Curvularia leaf spot.

【Keywords】 Bipolaris papendorfii; Isolation; Screening; Identification; Bacillus subtilis; Antagonism;


【Funds】 National Key Research and Development Program of China (2018YFD0200607-02) China Agriculture Research System (CARS-02)

Download this article


    [1] Chen XR, Nan ZB, Yang CD, et al. Morphological and biological characteristics of three Bipolaris species isolated from grasses in Huanxian grassland, Gansu province [J]. Acta Prataclturae Sinica, 2003, 12 (6): 86–92 (in Chinese)

    [2] Deng H. Taxonomic studies of the Genus Bipolaris Shoemaker (Hyphomycetes) in China [D]. Tai’an: Master’s Thesis of Shandong Agricultural University, 2002 (in Chinese)

    [3] Guo YJ. Phylogenetic analysis of the Genera Bipolaris and Curvularia in China [D]. Beijing: Master’s Thesis of Chinese Academy of Agricultural Sciences, 2016 (in Chinese)

    [4] Kuan CS, Yew SM, Toh YF, et al. Dissecting the fungal biology of Bipolaris papendorfii: from phylogenetic to comparative genomic analysis [J]. DNA Research, 2015, 22 (3): 219–232

    [5] Sun Y, Liu B. Current status of biological control of corn diseases in China [J]. Henan Agriculture, 2016 (26): 28 (in Chinese)

    [6] Li FH, Ye HZ, Wang YT, et al. The research progress of maize curvularia leaf spot disease [J]. Journal of Maize Sciences, 2004, 12 (2): 97–101, 107 (in Chinese)

    [7] Chang XM, Che ZQ, Gao JY. Research progress on prevent corn major diseases and insect pests using variety resistance [J]. Modern Agricultural Sciences and Technology, 2010 (2): 92–93 (in Chinese)

    [8] Ma BC. Functional analysis of Clg2p, a Ras protein gene of Curvularia lunata [D]. Daqing: Master’s Thesis of Heilongjiang Bayi Agricultural University, 2014 (in Chinese)

    [9] Chen J, Yan HH, Gao ZG, et al. Identification techniques for physiological differentiation of Curvularia lunata in maize [J]. Acta Phytopathologica Sinica, 2003, 33 (2): 121–125 (in Chinese)

    [10] Gu YF, Zhang YF, Zhang XP. Isolation and identification of one anti-Rhizoctonis solaniendophytic bacteria strain from corn and its antagonism and promoting research [J]. Microbiology China, 2008, 35 (8): 1240–1245 (in Chinese)

    [11] Chen MY, Xia YG, Chen WX. Occurrence and control of maize curvularia leaf spot disease [J]. Journal of Henan Agricultural Science, 2000 (7): 19–20 (in Chinese)

    [12] Guo YJ, Niu YC, Deng H. Bipolaris and Curvularia species associated with corn leaf spot in northern China [J]. Plant Protection, 2016, 42 (5): 39–46 (in Chinese)

    [13] Zhao XL, Niu YC, Deng H. A survey of pathogenic fungi on five common gramineous weeds in Henan Province [J]. Plant Protection, 2013, 39 (1): 128–132, 140 (in Chinese)

    [14] Geng RM, Zhang JP, Yu LQ. Isolation, identification and herbicidal activity of Bipolaris sp. and its safety on crops [J]. Acta Agriculturae Zhejiangensis, 2008, 20 (6): 446–450 (in Chinese)

    [15] Zhao XL, Hu ZJ, Song P, et al. Pathogenic fungi on Avena fatua L. in wheat field of Henan Province [J]. Chinese Agricultural Science Bulletin, 2016, 32 (15): 166–170 (in Chinese)

    [16] Wang GH, Raaijmakers JM. Antibiotics production by bacterial agents and its role in biological control [J]. Chinese Journal of Applied Ecology, 2004, 15 (6): 1100–1104 (in Chinese)

    [17] Li JZ, Wen CY. Research progress on bacteria for biological control of plant disease [J]. Journal of Henan Agricultural Sciences, 2015, 44 (10): 1–7 (in Chinese)

    [18] Yang XR, Liu SF, Sun SQ, et al. Research development on preventing soil-borne disease with anti-bacteria [J]. Tianjin Agricultural Sciences, 2008, 14 (4): 38–42 (in Chinese)

    [19] An X. Pesticide-degradation characteristics and degradation mechanism of biocontrol bacteria [D]. Jinan: Master’s Thesis of Shandong Normal University, 2010 (in Chinese)

    [20] Xu YJ, Liu HL, Liu XJ, et al. Advance in research of the bacteria biological control for plant diseases [J]. Soybean Science & Technology, 2011 (5): 18–22, 34 (in Chinese)

    [21] Luo WF. Screening of biocontrol strain to Rhizoctonia solani and preliminary research on the mechanisms [D]. Shenyang: Master’s Thesis of Shenyang Agricultural University, 2018 (in Chinese)

    [22] Qiu FB, Li YJ, Zhang XX, et al. Identification and antibiotic activity of endophytic bacterium strain ge21 from ginseng root [J]. Microbiology China, 2010, 37 (1): 43–47 (in Chinese)

    [23] Dong XZ, Cai MY. Manual of System Identification of Common Bacteria [M]. Beijing: Science Press, 2001 (in Chinese)

    [24] Kim SB, Yoon JH, Kim HG, et al. A phylogenetic analysis of the genus Saccharomonospora conducted with 16S rRNAgene sequences [J]. International Journal of Systematic Bacteriology, 1995, 45 (2): 351–356

    [25] Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees [J]. Molecular Biology and Evolution, 1987, 4 (4): 406–425

    [26] Yao ZS, Chen ZY, Chen ZY, et al. Genetically marking of natural biocontrol bacterium Bacillus subtilis strains with green fluorescent protein gene [J]. Chinese Journal of Biotechnology, 2003, 19 (5): 551–555 (in Chinese)

    [27] Qiu YX. Study on screening, identification and effect of efficient antagonistic bacteria against Botrytis cinerea [D]. Shijiazhuang: Master’s Thesis of Hebei Normal University, 2012 (in Chinese)

    [28] Kong J, Zhao BG, Wang WX, et al. On the inhibitory action of Bacillus subtilis B-903 strain’s antifungal substance on plant pathogenic fungi [J]. Acta Phytopathologica Sinica, 1995, 25 (1): 69–72 (in Chinese)

    [29] Zhai RH, Shang YK, Liu F, et al. Characteristics and inhibitory action of antifungal protein produced by Bacillus subtilis strain G8 [J]. Acta Phytophylacica Sinica, 2007, 34 (6): 592–596 (in Chinese)

    [30] Xing JS, Li R, Zhao L, et al. Purification, characterization and antagonism of an extracellular protease Bacillus subtilis strain T2 [J]. Acta Phytopathologica Sinica, 2008, 37 (4): 377–381 (in Chinese)

    [31] Deleu M, Paquot M, Nylander T. Effect of fengycin, a lipopeptide produced by Bacillus subtilis, on model biomembranes [J]. Biophysical Journal, 2008, 94 (7): 2667–2679

    [32] Feng JT, Han LR, Fan RJ, et al. Effects of cuminic acid on the growth and development of Phytophthora capsici leonian [J]. Scientia Agricultura Sinica, 2012, 45 (13): 2628–2635 (in Chinese)

This Article


CN: 11-1996/Q

Vol 47, No. 08, Pages 2417-2424

August 2020


Article Outline


  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion and conclusion
  • References