Screening and Identification of Antagonistic Bacterium against Banana Fusarium wilt and Its Biocontrol Effects

doi:10.13560/j.cnki.biotech.bull.1985.2025-0891

Abstract

Abstract:

Objective Banana Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (FOC), is a devastating soil-borne disease that seriously impacts banana production in terms of cultivation area, yield, and fruit quality. This study is aimed to screen effective biocontrol strains against FOC to facilitate the green and sustainable management of this disease. Method Antagonistic bacteria were isolated from the rhizosphere soil of banana plants. A potent isolate, GX-13, was selected and identified based on its morphological, physiological and biochemical characteristics, 16S rRNA gene sequence, and whole genome sequence, with subsequent annotation for potential biocontrol-related genes. Additionally, we assessed its plant-growth-promoting traits (phosphate solubilization, ammonia production, indole-3-acetic acid (IAA) synthesis) and antifungal properties (production of siderophores and cell wall-degrading enzymes). The efficacy of its sterile fermentation supernatant, volatile and non-volatile metabolites against FOC were evaluated in vitro, along with its inhibitory spectrum against other plant pathogenic fungi. Finally, the biocontrol efficacy and plant-growth-promoting ability of GX-13 were validated in a pot experiment. Result A bacterial strain, GX-13, showing antagonistic activity against FOC, was identified as Bacillus subtilis. GX-13 presented plant growth-promoting traits, including phosphate solubilization, ammonia production, indole-3-acetic acid synthesis, and antifungal functional characteristics such as secretion of fungal cell wall-degrading enzymes (protease, β-1,3-glucanase, cellulase) and siderophore production. The non-volatile metabolites of GX-13 caused structural damage to FOC hyphae, resulting in an inhibition rate of 76.85% and significantly reducing hyphal invasiveness. In contrast, its volatile metabolites and sterile fermentation supernatant showed lower inhibition rates (20.01% and 16.48%, respectively). GX-13 also demonstrated a broad antifungal spectrum, effectively inhibiting eight other phytopathogenic fungi, including Fusarium oxysporum f. sp. niveum, Fusarium oxysporum f. sp. lycopersici, Magnaporthe oryzae, Colletotrichum acutatum, Aspergillus carbonarius, Colletotrichum musae, Phyllosticta graminicola and Pseudopestalotiopsis theae. In pot experiments, GX-13 achieved control efficacies of 68.42 % against leaf yellowing and 50.00% against corm browning. Furthermore, it significantly promoted plant growth, with a highly significant enhancement in root development (P<0.01). Gene function annotation and secondary metabolite biosynthetic gene cluster analysis revealed that GX-13 harbored numerous genes involved in the synthesis fungal cell wall-degrading enzymes, antimicrobial compounds, and pathways responsible for plant growth promotion and pathogen antagonism. Conclusion The Bacillus subtilis strain GX-13 effectively controls banana Fusarium wilt through a synergistic mechanisms, including direct antifungal activity, broad-spectrum antagonism, and plant growth promotion.

Key words: banana Fusarium wilt, Fusarium oxysporum f. sp. cubense, Bacillus subtilis, antagonism, growth promotion

LIN Bao-mei, LI Shan-shan, LI Hai-ming, HONG Jia-min, ZHANG Shuai, WU Miao-hong, WU Wei-jian, WU Shui-jin. Screening and Identification of Antagonistic Bacterium against Banana Fusarium wilt and Its Biocontrol Effects[J]. Biotechnology Bulletin, 2026, 42(4): 272-286.

Figures/Tables 18

References 40

[1]	de Jesus Rocha A, da Silva Soares JM, dos Santos Nascimento F, et al. Improvements in the resistance of the banana species to Fusarium wilt: a systematic review of methods and perspectives [J]. J Fungi, 2021, 7(4): 249.
[2]	Ruan YN, Nong CH, Jintrawet A, et al. A smooth vetch (Vicia villosa Var.) strain endogenous to the broad-spectrum antagonist Bacillus siamensis JSZ06 alleviates banana wilt disease [J]. Front Plant Sci, 2024, 15: 1410197.
[3]	Li CL, Xiang DD, Yang S, et al. Fusarium wilt of banana latency and onset detection based on visible/near infrared spectral technology [J]. Agronomy, 2024, 14(12): 2994.
[4]	赵阳, 刘爽, 王志彪, 等. 香蕉枯萎病菌候选效应蛋白FoSSP80能抑制植物免疫反应 [J]. 热带生物学报, 2025, 16(1): 87-97.
	Zhao Y, Liu S, Wang ZB, et al. Functional characterization of candidate effector FoSSP80 in Fusarium oxysporum f. sp. cubense [J]. J Trop Biol, 2025, 16(1): 87-97.
[5]	张妙宜, 周登博, 起登凤, 等. 香蕉枯萎病综合防控研究进展 [J]. 中国科学: 生命科学, 2024, 54(10): 1843-1852.
	Zhang MY, Zhou DB, Qi DF, et al. Research progress on the integrated control of Fusarium wilt disease in banana [J]. Sci Sin Vitae, 2024, 54(10): 1843-1852.
[6]	李华平, 李云锋, 聂燕芳. 香蕉枯萎病的发生及防控研究现状 [J]. 华南农业大学学报, 2019, 40(5): 128-136.
	Li HP, Li YF, Nie YF. Research status of occurrence and control of Fusarium wilt of banana [J]. J South China Agric Univ, 2019, 40(5): 128-136.
[7]	Shen NK, Li SY, Li SY, et al. The siderophore-producing bacterium, Bacillus siamensis Gxun-6, has an antifungal activity against Fusarium oxysporum and promotes the growth of banana [J]. Egypt J Biol Pest Control, 2022, 32(1): 34.
[8]	Hasan M, Hossain M, Jiang D. New endophytic strains of Trichoderma promote growth and reduce clubroot severity of rapeseed (Brassica napus) [J]. PLoS One, 2023, 18(10): e0287899.
[9]	De Simone N, Capozzi V, Amodio ML, et al. Microbial-based biocontrol solutions for fruits and vegetables: recent insight, patents, and innovative trends [J]. Recent Pat Food Nutr Agric, 2021, 12(1): 3-18.
[10]	Bubici G, Kaushal M, Prigigallo MI, et al. Corrigendum: biological control agents against Fusarium wilt of banana [J]. Front Microbiol, 2019, 10: 1290.
[11]	Yin C, Casa Vargas JM, Schlatter DC, et al. Rhizosphere community selection reveals bacteria associated with reduced root disease [J]. Microbiome, 2021, 9(1): 86.
[12]	Guo YQ, Tian L, Zhu XY, et al. Antagonism of Bacillus velezensis ZGE166 against the pathogenic fungi causing corm rot disease in saffron (Crocus sativus L.) [J]. Microb Ecol, 2025, 88(1): 40.
[13]	Vibha R, Granada DL, Skariyachan S, et al. In vitro and in silico investigation deciphering novel antifungal activity of endophyte Bacillus velezensis CBMB205 against Fusarium oxysporum [J]. Sci Rep, 2025, 15: 684.
[14]	Qi D, Liu Q, Zou L, et al. Taxonomic identification and antagonistic activity of Streptomyces luomodiensis sp. nov. against phytopathogenic fungi [J]. Front Microbiol, 2024, 15: 1402653.
[15]	Hernández-Melchor DJ, Guerrero-Chávez AC, Ferrera-Rodríguez MR, et al. Cellulase and chitinase activities and antagonism against Fusarium oxysporum f.sp. cubense race 1 of six Trichoderma strains isolated from Mexican maize cropping [J]. Biotechnol Lett, 2023, 45(3): 387-400.
[16]	Solórzano R, Ramírez Maguiña HA, Johnson L, et al. Current progress in microbial biocontrol of banana Fusarium wilt: a systematic review [J]. Agronomy, 2025, 15(3): 619.
[17]	Bagy HMMK, Hassan EA, Nafady NA, et al. Efficacy of arbuscular mycorrhizal fungi and endophytic strain Epicoccum nigrum ASU11 as biocontrol agents against blackleg disease of potato caused by bacterial strain Pectobacterium carotovora subsp. atrosepticum PHY7 [J]. Biol Control, 2019, 134: 103-113.
[18]	吴巳雨. 长枝木霉HK16-1抗香蕉枯萎病活性物质的挖掘及抑菌效果分析 [D]. 武汉: 华中农业大学, 2024.
	Wu SY. Extraction of active substance from Trichoderma longibranchus HK16-1 against banana Fusarium wilt and analysis of antifungal effect [D]. Wuhan: Huazhong Agricultural University, 2024.
[19]	Samaras A, Roumeliotis E, Ntasiou P, et al. Bacillus subtilis MBI600 promotes growth of tomato plants and induces systemic resistance contributing to the control of soilborne pathogens [J]. Plants, 2021, 10(6): 1113.
[20]	张德政, 聂太礼, 杨军, 等. 棉花黄萎病高效拮抗细菌的筛选及其防治效果 [J]. 湖北农业科学, 2024, 63(12): 67-72.
	Zhang DZ, Nie TL, Yang J, et al. Screening and control effect of highly effective antagonistic bacteria against cotton verticillium wilt [J]. Hubei Agric Sci, 2024, 63(12): 67-72.
[21]	东秀珠, 蔡妙英. 常见细菌系统鉴定手册 [M]. 北京: 科学出版社, 2001.
	Dong XZ, Cai MY. Manual for common bacterial system identification [M]. Beijing: Science Press, 2001.
[22]	赵斌, 何绍江. 微生物学实验 [M]. 北京: 科学出版社, 2002.
	Zhao B, He SJ. Microbiological experiment [M]. Beijing: Science Press, 2002.
[23]	毕钰, 张琳, 余红凤, 等. 小麦赤霉病拮抗菌JB7的拮抗活性及鉴定 [J]. 微生物学报, 2024, 64(5): 1580-1592.
	Bi Y, Zhang L, Yu HF, et al. Antagonistic activity and identification of antagonistic bacteria JB7 against Fusarium head blight [J]. Acta Microbiol Sin, 2024, 64(5): 1580-1592.
[24]	杨学宇, 谭琳, 沈程文, 等. 茶轮斑病拮抗细菌kc-16的抑菌性能及田间防效 [J]. 微生物学通报, 2024, 51(11): 4560-4573.
	Yang XY, Tan L, Shen CW, et al. Antimicrobial activity and field control effect of an antagonistic bacterial strain kc-16 against tea gray blight [J]. Microbiol China, 2024, 51(11): 4560-4573.
[25]	李文博, 张新祺, 赵亚婷, 等. 采前喷施壳寡糖对采后西梅黑斑病的控制 [J]. 食品科学, 2025, 46(7): 283-291.
	Li WB, Zhang XQ, Zhao YT, et al. Control of postharvest prune black spot disease by preharvest chitosan oligosaccharide spraying [J]. Food Sci, 2025, 46(7): 283-291.
[26]	黎晓丹,方献平,李水根,等.草莓果生炭疽菌效应子基因CfE4对产孢量及菌丝穿透能力的影响[J/OL].分子植物育种,1-11[2025-12-12].,
	Fang XP, Li SG, et al. The effect of effector gene CfE4 on conidia production and penetration of Colletotrichum fructicola in strawberry[J/OL]. [2025-5-27]. .
[27]	Hyder S, Gondal AS, Rizvi ZF, et al. Characterization of native plant growth promoting rhizobacteria and their anti-oomycete potential against Phytophthora capsici affecting chilli pepper (Capsicum annum L.) [J]. Sci Rep, 2020, 10: 13859.
[28]	甘林, 代玉立, 刘晓菲, 等. 香蕉枯萎病高效拮抗土著细菌的筛选及其防效 [J]. 西北农林科技大学学报: 自然科学版, 2024, 52(6): 95-105.
	Gan L, Dai YL, Liu XF, et al. Screening and control efficacies of highly effective antagonistic native bacteria against banana Fusarium wilt [J]. J Northwest A F Univ Nat Sci Ed, 2024, 52(6): 95-105.
[29]	Wang JR, Qin S, Fan RD, et al. Plant growth promotion and biocontrol of leaf blight caused by Nigrospora sphaerica on passion fruit by endophytic bacillus subtilis strain GUCC4 [J]. J Fungi, 2023, 9(2): 132.
[30]	Sun WL, Shahrajabian MH, Soleymani A. The roles of plant-growth-promoting rhizobacteria (PGPR)-based biostimulants for agricultural production systems [J]. Plants, 2024, 13(5): 613.
[31]	黄勋, 刘霞, 邓琳梅, 等. 马铃薯疮痂病生防菌1X1Y的鉴定及其生防促生特性研究 [J]. 园艺学报, 2025, 52(1): 229-246.
	Huang X, Liu X, Deng LM, et al. Identification and study on its biocontrol and growth promoting properties of Bacillus velezensis 1X1Y against potato common scab [J]. Acta Hortic Sin, 2025, 52(1): 229-246.
[32]	Yang P, Yuan P, Liu WS, et al. Plant growth promotion and plant disease suppression induced by Bacillus amyloliquefaciens strain GD4a [J]. Plants, 2024, 13(5): 672.
[33]	邓玉莲, 谭琳, 吉进军, 等. 茶树根腐病拮抗真菌的筛选、鉴定及防病促生特性研究 [J]. 中国生物防治学报, 2024, 40(4): 914-926.
	Deng YL, Tan L, Ji JJ, et al. Screening and identification of antagonistic fungi against root rot of tea plants and their effect on disease control and plant growth promotion [J]. Chin J Biol Control, 2024, 40(4): 914-926.
[34]	Dutilloy E, Arias AA, Richet N, et al. Bacillus velezensis BE2 controls wheat and barley diseases by direct antagonism and induced systemic resistance [J]. Appl Microbiol Biotechnol, 2024, 108(1): 64.
[35]	Dai MM, Liu R, Jiang H, et al. Volatile organic compounds of Bacillus pumilus strain S1-10 exhibit fumigant activity against Meloidogyne incognita [J]. Plant Dis, 2023, 107(10): 3057-3063.
[36]	Guevara-Avendaño E, Bravo-Castillo KR, Monribot-Villanueva JL, et al. Diffusible and volatile organic compounds produced by avocado rhizobacteria exhibit antifungal effects against Fusarium kuroshium [J]. Braz J Microbiol, 2020, 51(3): 861-873.
[37]	刘高强, 李新鹏, 刘文钊, 等.一株高效降解血液蛋白的枯草芽孢杆菌NWMCC0137全基因组测序及分析 [J]. 江苏农业学报, 2023, 39(7): 1460-1471.
	Liu GQ, Li XP, Liu WZ, et al. Whole genome sequencing and analysis of Bacillus subtilis NWMCC0137, an efficient blood protein degrading strain [J]. Jiangsu J Agric Sci, 2023, 39(7): 1460-1471.
[38]	万青. 一株贝莱斯芽孢杆菌对香蕉枯萎病生防作用的研究 [D]. 海口: 海南大学, 2021.
	Wan Q. Study on the biocontrol effect of a Bacillus velezensis on banana Fusarium wilt [D]. Haikou: Hainan University, 2021.
[39]	李恒, 畅文军, 陈汉清, 等. 香蕉枯萎镰刀菌4号生理小种mon1基因敲除转化子的表型分析及致病力测定 [J]. 热带生物学报, 2019, 10(2): 127-134.
	Li H, Chang WJ, Chen HQ, et al. Functional analysis of the mon1 gene in Fusarium oxysporum f.sp. cubense Race 4 [J]. J Trop Biol, 2019, 10(2): 127-134.
[40]	王田, 陈代朋, 高雅, 等. 香蕉枯萎病菌效应蛋白研究进展 [J]. 农业生物技术学报, 2022, 30(8): 1614-1621.
	Wang T, Chen DP, Gao Y, et al. Research progress on the effector of Fusarium oxysporum f.sp. cubense [J]. J Agric Biotechnol, 2022, 30(8): 1614-1621.

评价指标 Evaluation indicator	病害等级 Pathogen severity grading	病害描述 Evaluation content description
叶片黄化度 Leaf yellowing degree	0	叶片无症状
	1	下部1-2片叶黄化，黄化面积≤25%
	3	3-4片叶黄化，25%<黄化面积≤50%
	5	外围叶片黄化，黄化面积>50%，叶片萎蔫
	7	大部分叶片黄化、萎蔫，仅叶心正常
	9	植物萎蔫、枯萎死亡
球茎褐变度 Bulb browning degree	0	球茎组织白色，无褐变
	1	球茎褐变面积≤25%
	3	25%<球茎褐变面积≤50%
	5	50%<球茎褐变面积≤75%
	7	球茎褐变面积>75%

评价指标 Evaluation indicator	病害等级 Pathogen severity grading	病害描述 Evaluation content description
叶片黄化度 Leaf yellowing degree	0	叶片无症状
	1	下部1-2片叶黄化，黄化面积≤25%
	3	3-4片叶黄化，25%<黄化面积≤50%
	5	外围叶片黄化，黄化面积>50%，叶片萎蔫
	7	大部分叶片黄化、萎蔫，仅叶心正常
	9	植物萎蔫、枯萎死亡
球茎褐变度 Bulb browning degree	0	球茎组织白色，无褐变
	1	球茎褐变面积≤25%
	3	25%<球茎褐变面积≤50%
	5	50%<球茎褐变面积≤75%
	7	球茎褐变面积>75%

试验项目 Tested item		结果 Results	试验项目 Item tested		结果 Results
接触酶反应 Catalase reaction		+	V-P试验 Voges Proskauer test		+
明胶液化 Gelatine liquefication		+	甲基红试验 Methyl red test		-
吲哚试验 Indole test		-	苯丙氨酸脱氨酶 Amphetanine dehydrogenase reaction		-
淀粉水解 Starch hydrolysis		+	产氨试验 Ammonia production test		+
不同碳源 Different carbon sources	葡萄糖 Glucose	+	硝酸盐还原实验 Nitrate reduction reaction		+
	蔗糖 Sucrose	+	柠檬酸盐试验 Citrate test		+
	D-甘露醇 D-mannitol	+	不同氮源 Different nitrogen sources	硝酸钾 KNO₃	+
	D-麦芽糖 D-maltdust	+	不同氮源 Different nitrogen sources	硫酸铵（NH₄）₂SO₄	+

试验项目 Tested item		结果 Results	试验项目 Item tested		结果 Results
接触酶反应 Catalase reaction		+	V-P试验 Voges Proskauer test		+
明胶液化 Gelatine liquefication		+	甲基红试验 Methyl red test		-
吲哚试验 Indole test		-	苯丙氨酸脱氨酶 Amphetanine dehydrogenase reaction		-
淀粉水解 Starch hydrolysis		+	产氨试验 Ammonia production test		+
不同碳源 Different carbon sources	葡萄糖 Glucose	+	硝酸盐还原实验 Nitrate reduction reaction		+
	蔗糖 Sucrose	+	柠檬酸盐试验 Citrate test		+
	D-甘露醇 D-mannitol	+	不同氮源 Different nitrogen sources	硝酸钾 KNO₃	+
	D-麦芽糖 D-maltdust	+	不同氮源 Different nitrogen sources	硫酸铵（NH₄）₂SO₄	+

菌株 Strain	ANI （%）
B. subtilis ATCC6051	99.35
B. cabrialesii TE3	92.41
B. tequilensis ATCC BAA-819	91.71
B. halotolerans ZB201702	87.43
B. velezensis FZB42	77.24
B. amyloliquefaciens ATCC23350	77.02
B. paralicheniformis KJ-16	72.80
B. australimaris PWG	71.01
B. pumilus ATCC 7061	70.99