|
|
|
| Screening of a Bacillus Strain GXMZ-G3 Exhibiting Induced Systemic Resistance and Its Biocontrol Efficacy Against Banana Fusarium Wilt |
| YIN Dou-Dou1,*, SHEN Nai-Kun1,*, HUANG Xiao-Tian1, LUO Yan-Xin1, HUANG Yi-Hui1, LI Bao-Shen2, ZHANG Hong-Yan1,** |
1 Guangxi Key Laboratory for Polysaccharide Materials and Modification/School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning 530006, China;
2 Biotechnology Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530006, China |
|
|
|
|
Abstract Banana Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), represents a severe threat to global banana (Musa nana) cultivation industry. To develop environmentally sustainable strategies for managing this devastating disease, this study aimed to screen and characterize bacterial biocontrol agents capable of inducing systemic resistance in banana plants. Candidate strains were selected based on their ability to enhance the activity of key defense-related enzymes, including polyphenol oxidase (PPO), peroxidase (POD), and superoxide dismutase (SOD), in banana seedlings. The most promising strain was rigorously identified using integrated taxonomic approaches, encompassing morphological, physiological, and biochemical characterization, as well as phylogenetic analysis of the 16S rRNA gene sequence. Subsequently, its optimal culture conditions were systematically optimized. Antagonistic activity of the selected strain against Foc TR4 was evaluated through dual-culture plate assays and co-cultivation experiments with conidial suspensions to assess inhibition of mycelial growth and spore germination. Its capacity to induce systemic resistance in banana plants was further validated via root drench inoculation under controlled conditions. The results revealed that a single isolate, designated GXMZ-G3, was identified as Bacillus sp. This strain effectively induced systemic resistance in banana plants. In vitro assays demonstrated that GXMZ-G3 exhibited a 62.63% inhibition rate against Foc TR4 mycelial growth and suppressed conidial germination by 85.66%. Microscopic observation revealed severe morphological abnormalities in the inhibited Foc TR4 hyphae, including twisting, knotting, and fragmentation. Following root drench application of the GXMZ-G3 suspension, the activities of key defense-related enzymes (PPO, POD, and SOD) were elevated in banana leaves. Moreover, hydrogen peroxide (H2O2) accumulation and callose deposition were markedly enhanced in treated leaves. Jasmonic acid (JA) levels peaked at 0.48 ng/mL at 12 h post-inoculation, representing a 20.00% increase relative to the untreated control. In pot experiments, GXMZ-G3 achieved an 89.48% disease control efficacy against banana Fusarium wilt. In summary, the biocontrol strain GXMZ-G3 exerts a dual mode of action. It directly inhibited Foc TR4 hyphal growth and spore germination, while simultaneously priming systemic resistance in banana plants. These findings highlight GXMZ-G3 as a promising microbial resource for the biological control of banana Fusarium wilt disease.
|
|
Received: 19 November 2025
|
|
|
|
Corresponding Authors:
**hongyanzhang2008@163.com
|
| About author:: * These authors contributed equally to this work |
|
|
|
[1] 布坎南R E, 吉本斯N E. 1984. 伯杰细菌鉴定手册[M]. 第八版, 北京: 科学出版社, pp. 85-86.
(Buchanan R E, Gibbons N E.1984. Bergey's manual of determinative bacteriology[M]. 8th ed. Science Press, Beijing, China, pp. 85-86.)
[2] 蔡文涌, 方香玲. 2024. 植物病原真菌尖孢镰刀菌致病因子研究进展[J]. 分子植物育种, 22(24): 8270-8278.
(Cai W Y,Fang X L.2024. Research progress on pathogenic factors of different Fusarium oxysporum[J]. Molecular Plant Breeding, 22(24): 8270-8278.)
[3] 曹雨. 2021. 基于转录组研究解淀粉芽孢杆菌SQR9诱导玉米茎腐病系统抗性的机制[D]. 硕士学位论文, 南京农业大学, 导师: 高夕全, pp. 28-29.
(Cao Yu.2021. Study on the mechanism of Bacillus amyloliquefaciens SQR9-induced systemic resistance to Gibberella stalk rot based on the transcriptome[D]. Thesis for M.S., Nanjing Agricultural University, Supervisor: Gao X X, pp. 28-29.)
[4] 董爱菊. 2021. 类芽孢杆菌QHZ11对马铃薯黑痣病的生防机制及对马铃薯的促生效果研究[D]. 硕士学位论文, 甘肃农业大学, 导师: 邱慧珍, pp. 26-31.
(Dong A J.2021. Studies on the biocontrol mechanism of Paenibacillus jamilae QHZ11 on black scurf disease of potato and its promoting effect for potato plants[D]. Thesis for M.S., Gansu Agricultural University, Supervisor: Qiu H Z, pp. 26-31.)
[5] 杜婵娟, 杨迪, 潘连富, 等. 2020. 香蕉枯萎病生防菌肥的优化及其防病促生效果研究[J]. 中国生物防治学报, 36(3): 396-404.
(Du C J, Yang D, Pan L Fu, et al.2020. Bio-fertilizer optimization and its control effect on banana Fusarium wilt[J]. Chinese Journal of Biological Control, 36(3): 396-404.)
[6] 范宗强, 冯靖涵, 郑丽雪, 等. 2024. 枯草芽孢杆菌B579对黄瓜枯萎病的防治及其诱导抗性研究[J]. 生物技术通报, 40(7): 226-234.
(Fan Z Q, Feng J H, Zheng L X, et al.2024. Study on the control and induced resistance in cucumber with Bacillus subtilis B579 against cucumber Fusarium wilt[J]. Biotechnology Bulletin, 40(7): 226-234.)
[7] 甘林, 代玉立, 刘晓菲, 等. 2024. 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), 52(6): 95-105.
(Gan L, Dai Y L, Liu X F, et al.2024. Screening and control efficacies of highly effective antagonistic native bacteria against banana Fusarium wilt[J]. Journal of Northwest A＆F University (Natural Science Edition), 52(6): 95-105.)
[8] 李丽丽, 曾广宇, 王继春, 等. 2024. 油茶根腐病内生拮抗促生细菌筛选及诱导系统抗性研究[J]. 中国油料作物学报, 46(6): 1390-1397.
(Li L L, Zeng G Y, Wang J C, et al.2024. Screening and systematic resistance induction of endogenous antagonistic growth-promoting bacteria against Camellia oleifera root rot[J]. Chinese Journal of Oil Crop Sciences, 46(6): 1390-1397.)
[9] 李枢妍, 阳黎恒, 肖雪婷, 等. 2021. 一株香蕉枯萎病拮抗菌的筛选、鉴定及生防效果研究[J]. 南方农业学报, 52(7): 1826-1834.
(Li S Y, Yang L H, Xiao X T, et al.2021. Screening, identification and biocontrol effect of antagonistic bacteria against banana Fusarium wilt[J]. Journal of Southern Agriculture, 52(7): 1826-1834.)
[10] 李小芳, 张志良. 2016. 植物生理学实验指导[M]. 高等教育出版社, 北京, pp. 67-69.
(Li X F, Zhang Z L.2016. Experimental instruction of plant physiology[M]. Higher education press, Beijing, China, pp. 67-69.)
[11] 陆君铭, 吉春喜, 郭健杰, 等. 2025. 一株耐镉促生细菌Achromobacter sp. A81的分离鉴定及特性[J]. 微生物学报, 65(1): 90-105.
(Lu J M, Ji C X, Guo J J, et al.2025. Isolation, identification, and characterization of a cadmium-tolerant bacterium Achromobacter sp. A81 with plant growth-promoting effect[J]. Acta Microbiologica Sinica, 65(1): 90-105.)
[12] 沙月霞, 张昂, 伍顺华, 等. 2020. 假单胞菌S149对水稻防御酶的诱导及定殖能力[J]. 中国植保导刊, 40(7): 10-16.
(Sha Y X, Zhang A, Wu S H, et al.2020. Colonization ability of Pseudomonas S149 and induction of defense-related enzymes in rice[J]. China Plant Protection, 40(7): 10-16.)
[13] 苏琴, 谢玲, 陈艳露, 等. 2019. 香蕉枯萎病生防菌绿头枝孢菌LS1的筛选鉴定[J]. 微生物学通报, 46(12): 3248-3256.
(Su Q, Xie L, Chen Y L, et al.2019. Screen and identification of biocontrol strain Cladosporium chlorocephalum LS1 against banana Fusarium wilt[J]. Microbiology China, 46(12): 3248-3256.)
[14] 王鹏. 2022. 马铃薯黑痣病菌拮抗细菌的筛选及其生防机制研究[D]. 硕士学位论文, 黑龙江八一农垦大学, 导师: 台莲梅, pp. 15-17.
(Wang P.2022. Screening of antagonistic bacteria against potato black scurf and study on its biocontrol mechanism[D]. Thesis for M.S., Heilongjiang Bayi Agricultural University, Supervisor: Tai L M, pp. 15-17.)
[15] 王丽, 王万兴, 索海翠, 等. 2019. 马铃薯块茎酶促褐变及与相关生理指标的关系[J]. 园艺学报, 46(8): 1519-1530.
(Wang L, Wang W X, Suo H C, et al.2019. The relationship between enzymatic browning and relevant physiological index of potato tubers[J]. Acta Horticulturae Sinica, 46(8): 1519-1530.)
[16] 吴耿尉. 2018. 解淀粉芽孢杆菌SQR9诱导植物系统抗性及其激发子鉴定[D]. 博士学位论文, 南京农业大学, 导师: 张瑞福, pp. 67-69.
(Wu G W.2018. Identification of elicitors to induced plant systemic resistance by Bacillus amyloliquefaciens SQR9[D]. Thesis for Ph.D., Nanjing Agricultural University, Supervisor: Zhang R F, pp. 67-69.)
[17] 徐玉. 2019. 木霉NJAU 4742对植物生长和免疫反应的诱导作用及其机理研究[D]. 博士学位论文, 南京农业大学, 导师: 沈其荣, pp. 1-2.
(Xu Y.2019. Induction of plant growth and immune response by Trichoderma guizhouense NJAU 4742 and its mechanism[D]. Thesis for Ph.D., Nanjing Agricultural University, Supervisor: Shen Q R, pp. 1-2.)
[18] 殷豆豆, 黄宏智, 宋超东, 等. 2024. 广西小花老鼠簕可培养细菌多样性及促生、防病、产酶活性分析[J]. 南方农业学报, 55(4): 973-984.
(Yin D D, Huang H Z, Song C D, et al.2024. Diversity of culturable endophytic bacteria in Acanthus ebracteatus of Guangxi and their growth-promoting, biocontrol effect and enzyme production activity[J]. Journal of Southern Agriculture, 55(4): 973-984.)
[19] 宛甜甜. 2023. 解淀粉芽孢杆菌SN16-1对水稻稻瘟病的防治作用及其诱抗机理研究[D]. 硕士学位论文, 华东理工大学, 导师: 王伟, 蒋贤权, pp. 41-49.
(Wan T T.2023. Control of rice blast by Bacillus amyloliquefaciens SN16-1 and its resistance-inducing mechanism[D]. Thesis for M.S., East China University of Science and Technology, Supervisor: Wang W, Jiang X Q, pp. 41-49.)
[20] 周林. 2011. 枯草芽孢杆菌TR21对香蕉枯萎病生防机制研究[D]. 硕士学位论文, 华中农业大学, 导师: 程萍, pp. 1-3.
(Zhou L.2011. Investigation on the biocontrol mechanism of Bacillus subtilis strain TR21 against banana Fusarium wilt[D]. Thesis for M.S., Huazhong Agricultural University, Supervisor: Cheng P, pp. 1-3.)
[21] 周林, 程萍, 喻国辉, 等. 2021. 枯草芽孢杆菌TR21对香蕉抗病相关酶活的诱导作用[J]. 中国农学通报, 27(2): 185-190.
(Zhou L, Cheng P, Yu G H, et al.2021. The induction of Bacillus subtilis strain TR21 on related enzymes of disease resistance of banana seedling[J]. Chinese Agricultural Science Bulletin, 27(2): 185-190.)
[22] 周维, 李佳林, 黄素梅, 等. 2020. 广西香蕉枯萎病发生现状及防控措施[J]. 福建农业科技, 51(5): 46-50.
(Zhou W, Li J L, Huang S M, et al.2020. Occurrence status of banana Fusarium wilt in Guangxi and its prevention and control measures[J]. Fujian Agricultural Science and Technology, 51(5): 46-50.)
[23] 邹红梅. 2019. 微波、乙醇处理对鲜切马铃薯酶促褐变的抑制效果研究[D]. 硕士学位论文, 兰州理工大学, 导师: 王雅, pp. 8-10.
(Zou H M.2019. Effect of microwave and ethanol treatment on enzymatic browning of fresh-cut potatoes[D]. Thesis for M.S., Lanzhou University of Technology, Supervisor: Wang Y, pp. 8-10.)
[24] Andleeb Z, Anant N R, Mukesh M, et al.2021. Efficiency of microbial bio-agents as elicitors in plant defense mechanism under biotic stress: A review[J]. Current Research in Microbial Sciences, 2(1): 100054.
[25] Chen C, Zhang X H, Wei X, et al.2022. Postharvest biological control of Botrytis cinerea and the mechanisms underlying the induction of disease resistance in grapes by Lactobacillus plantarum CM-3[J]. Biological Control, 172(5): 104982.
[26] Droby S, Wisniewski M.2018. The fruit microbiome: A new frontier for postharvest biocontrol and postharvest biology[J]. Postharvest Biology and Technology, 140(1): 107-112.
[27] Gong A, Li H, Yuan Q, et al.2015. Antagonistic mechanism of iturin A and plipastatin A from Bacillus amyloliquefaciens S76-3 from wheat spikes against Fusarium graminearum[J]. PLOS ONE, 10(12): e0116871.
[28] Gupta R S, Patel S, Saini N, et al.2020. Robust demarcation of 17 distinct Bacillus species clades, proposed as novel Bacillaceae genera, by phylogenomics and comparative genomic analyses: Description of Robertmurraya kyonggiensis sp. nov. and proposal for an emended genus Bacillus limiting it only to the members of the Subtilis and Cereus clades of species[J]. International Journal of Systematic and Evolutionary Microbiology, 70(11): 5753-5798.
[29] Hong S W, Kwon S W, Kim S J, et al.2014. Bacillus oryzaecorticis sp. nov., a moderately halophilic bacterium isolated from rice husks[J]. International Journal of Systematic and Evolutionary Microbiology, 64(8): 2786-2791.
[30] Jinal N H, Amaresan N.2020. Evaluation of biocontrol Bacillus species on plant growth promotion and systemic-induced resistant potential against bacterial and fungal wilt-causing pathogens[J]. Archives of Microbiology, 202(7): 1785-1794.
[31] Li S Y, Ma J P, Li S Y, et al.2022. Comparative transcriptome analysis unravels the response mechanisms of Fusarium oxysporum f. sp. cubense to a biocontrol agent, Pseudomonas aeruginosa Gxun-2[J]. International Journal of Molecular Sciences, 23(24): 15432.
[32] Niu D D, Liu H X, Jiang C H, et al.2011. The plant growth-promoting rhizobacterium Bacillus cereus AR156 induces systemic resistance in Arabidopsis thaliana by simultaneously activating salicylate- and jasmonate/ethylene-dependent signaling pathways[J]. Molecular Plant-microbe Interactions, 24(5): 533-542.
[33] Polina C T, Eirini-Evangelia T, Costas D, et al.2023. Compatible consortium of endophytic Bacillus halotolerans strains Cal.l.30 and Cal.f.4 promotes plant growth and induces systemic resistance against Botrytis cinerea[J]. Biology, 12(6): 779.
[34] Raymaekers K, Ponet L, Holtappels D, et al.2020. Screening for novel biocontrol agents applicable in plant disease management-a review[J]. Biological Control, 144(3): 104240.
[35] Saleem D, Zuhra Z, Akhtar W, et al.2020. Salicylic acid and H2O2 induce PPO derived GUS expression in Arabidopsis[J]. Russian Journal of Plant Physiology, 67(5): 822-826.
[36] Sampathkumar A, Aiyanathan K E A, Nakkeeran S, et al.2023. Multifaceted Bacillus spp. for the management of cotton bacterial blight caused by Xanthomonas citri pv. malvacearum[J]. Biological Control, 177(1): 105123.
[37] Wang M Y, Zhang Y K, Cai H B, et al.2024. A new biocontrol agent Bacillus velezensis SF334 against rubber tree fungal leaf anthracnose and its genome analysis of versatile plant probiotic traits[J]. Journal of Fungi, 10(2): 158.
[38] Ye S, Yan R, Li X W, et al.2022. Biocontrol potential of Pseudomonas rhodesiae GC-7 against the root-knot nematode Meloidogyne graminicola through both antagonistic effects and induced plant resistance[J]. Frontiers in Microbiology, 13(1): 1025727.
[39] Yun T Y, Jing T, Zhou D B, et al.2022. Potential biological control of endophytic Streptomyces sp. 5-4 against Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense tropical race 4[J]. Phytopathology, 112(9): 1877-1885. |
|
|
|
