Potentially biological control of Bacillus spp. against Fusarium keratoplasticum Geiser et al. 2013 causing wilt in pico chrysanthemum
Main Article Content
Abstract
Fusarium wilt, caused by Fusarium spp., is considered one of the most serious diseases affecting pico chrysanthemum (Chrysanthemum spp.), leading to significant yield and quality losses. This study was conducted to identify the fungal pathogen causing wilt disease in pico chrysanthemum and to assess the antifungal potential of Bacillus spp. under in vitro conditions. The fungal isolate FC1 was recovered from infected plants and identified as Fusarium keratoplasticum through analysis of internal transcribed spacer (ITS) gene sequences together with morphological characteristics. Its pathogenicity was confirmed in accordance with Koch’s postulates. All 41 Bacillus isolates exhibited antifungal activity against F. keratoplasticum in dual culture assays, with mycelial growth inhibition ranging from 20.0% to 52.2%. Volatile organic compounds (VOCs) and cell-free supernatants (CFS) produced by six promising isolates (CB3, CB10, CB31, CB32, CB35, and CB36) markedly suppressed both mycelial growth and spore germination of fungal isolate FC1. These findings suggest that the selected Bacillus isolates have strong potential as biological control agents for managing F. keratoplasticum-induced wilt in pico chrysanthemum.
Article Details
Conflict of Interest

© 2026 The authors. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License.
References
Al-Samarrai, T. H., & Schmid, J. (2000). A simple method for extraction of fungal genomic DNA. Letters in Applied Microbiology, 30(1), 53-56. https://doi.org/10.1046/j.1472-765x.2000.00664.x
Aslam, S., Baber, M., Naqqash, T., Javed, M., Bredenbruch, S., Grundler, F. M. W., & Schleker, A. S. S. (2025). Rhizospheric Bacillus isolates control Fusarium wilt on cotton and enhance plant biomass and root development. Frontiers in Microbiology, 16, 1580937. https://doi.org/10.3389/fmicb.2025.1580937
Bahadur, A. (2022). Current status of Fusarium and their management strategies. IntechOpen.
Balamurugan, A., Ashajyothi, M., Shanu, K., Charishma, K., Varun, H., Gunjeet, K., & Kumar, A. (2024). Chrysanthemum wilt caused by Fusarium incarnatum: Etiology unveiled through polyphasic taxonomic methods. Physiological and Molecular Plant Pathology, 129, 102214. https://doi.org/10.1016/j.pmpp.2023.102214
Chandrasekaran, M., Paramasivan, M., & Sahayarayan, J. J. (2022). Microbial volatile organic compounds: an alternative for chemical fertilizers in sustainable agriculture development. Microorganisms, 11(1), 42. https://doi.org/10.3390/microorganisms11010042
Chaves-López, C., Serio, A., Gianotti, A., Sacchetti, G., Ndagijimana, M., Ciccarone, C., Stellarini, A., Corsetti, A., & Paparella, A. (2015). Diversity of food-borne Bacillus volatile compounds and influence on fungal growth. Journal of Applied Microbiology, 119, 487–499. https://doi.org/10.1111/jam.12847
Chehri, K., Salleh, B., & Zakaria, L. (2015). Morphological and phylogenetic analysis of Fusarium solani species complex in Malaysia. Microbial Ecology, 69(3), 457-471. https://doi.org/10.1007/s00248-014-0494-2
Chen, X., Shu, Y., Luo, M., Xiang, M., Huang, Y., Zhang, W., & Dong, Z. (2020). Fusarium wilt of imperial chrysanthemum (Chrysanthemum morifolium) caused by Fusarium oxysporum in China. Plant Disease, 103(4), 985. https://doi.org/10.1094/PDIS-10-19-2140-PDN
Chen, Z., Wang, Z., & Xu, W. (2024). Bacillus velezensis WB induces systemic resistance in watermelon against Fusarium wilt. Pest Management Science, 80(3), 1423–1434. https://doi.org/10.1002/ps.7873
Fujita, S. I., Senda, Y., Nakaguchi, S., & Hashimoto, T. (2001). Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains. Journal of Clinical Microbiology, 39(10), 3617-3622. https://doi.org/10.1128/JCM.39.10.3617-3622.2001
Ghazala, I., Chiab, N., Saidi, M. N., & Gargouri-Bouzid, R. (2022). Volatile organic compounds from Bacillus mojavensis I4 promote plant growth and inhibit phytopathogens. Physiological and Molecular Plant Pathology, 121, 101887. https://doi.org/10.1016/j.pmpp.2022.101887
Han, J. H., Shim, H., Shin, J. H., & Kim, K. S. (2015). Antagonistic activities of Bacillus spp. strains isolated from tidal flat sediment towards anthracnose pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea. The Plant Pathology Journal, 31(2), 165-175. https://doi.org/10.5423/PPJ.OA.03.2015.0036
Hao, D. C., Song, Y., Xiao, P., Zhong, Y., Wu, P., & Xu, L. (2022). The genus Chrysanthemum: Phylogeny, biodiversity, phytometabolites, and chemodiversity. Frontiers in Plant Science, 13, 973197. https://doi.org/10.3389/fpls.2022.973197
Iqbal, O., Syed, R. N., Rajput, N. A., Wang, Y., Lodhi, A. M., Khan, R., Jibril, S. M., Atiq, M., & Li, C. (2024). Antagonistic activity of two Bacillus strains against Fusarium oxysporum f. sp. capsici (FOC-1) causing Fusarium wilt and growth promotion activity of chili plant. Frontiers in Microbiology, 15, 1388439. https://doi.org/10.3389/fmicb.2024.1388439
James, J. E., Santhanam, J., Zakaria, L., Mamat Rusli, N., Abu Bakar, M., Suetrong, S., Sakayaroj, J., Abdul Razak, M. F., Lamping, E., & Cannon, R. D. (2022). Morphology, phenotype, and molecular identification of clinical and environmental Fusarium solani species complex isolates from Malaysia. Journal of Fungi (Basel, Switzerland), 8(8), 845. https://doi.org/10.3390/jof8080845
Khan, N., Maymon, M., & Hirsch, A. M. (2017). Combating Fusarium infection using Bacillus-based antimicrobials. Microorganisms, 5(4), 75. https://doi.org/10.3390/microorganisms5040075
Kim, H. S., Kim, J. Y., Lee, S. M., Park, H. J., Lee, S. H., Jang, J. S., & Lee, M. H. (2019). Isolation and characterization of various strains of Bacillus sp. having antagonistic effect against phytopathogenic fungi. Microbiology and Biotechnology Letters, 47(4), 603-613. https://doi.org/10.4014/mbl.1904.04001
Lien, N. T., Oanh, N. T. P., & Khoa, N. D. (2023). Study on characterization of Bacillus amyloliquefaciens VL4.6 as a biocontrol agent against Colletotrichum lagenarium causing anthracnose on cucumber. Vietnam Journal of Agriculture and Rural Development, 10, 34-41.
Liu, L., Jin, Y., Chen, M., Lian, H., Liu, Y., Yin, Q., and Wang, H. (2023a). A Study of Soil-Borne Fusarium Wilt in Continuous Cropping Chrysanthemum Cultivar 'Guangyu' in Henan, China. Journal of Fungi (Basel, Switzerland), 10(1), 14. https://doi.org/10.3390/jof10010014
Liu, L., Jin, X., Lu, X., Guo, L., Lu, P., Yu, H., & Lv, B. (2023b). Mechanisms of surfactin from Bacillus subtilis SF1 against Fusarium foetens: a novel pathogen inducing potato wilt. Journal of Fungi (Basel, Switzerland), 9(3), 367. https://doi.org/10.3390/jof9030367
Mekapogu, M., Kwon, O. K., Song, H. Y., & Jung, J. A. (2022). Towards the improvement of ornamental attributes in chrysanthemum: recent progress in biotechnological advances. International Journal of Molecular Sciences, 23(20), 12284. https://doi.org/10.3390/ijms232012284
Myo, E. M., Liu, B., Ma, J., Shi, L., Jiang, M., Zhang, K., & Ge, B. (2019). Evaluation of Bacillus velezensis NKG-2 for bio-control activities against fungal diseases and potential plant growth promotion. Biological Control, 134, 23–31. https://doi.org/10.1016/j.biocontrol.2019.03.017
Naqvi, S. A. H., Farhan, M., Ahmad, M., Kiran, R., Shahbaz, M., Abbas, A., Hakim, F., Shabbir, M., Tan, Y. S., & Sathiya Seelan, J. S. (2025). Fungicide resistance in Fusarium species: exploring environmental impacts and sustainable management strategies. Archives of Microbiology, 207(2), 31. https://doi.org/10.1007/s00203-024-04219-6
Sharma, L., Oliveira, I., Torres, L., & Marques, G. (2018). Entomopathogenic fungi in Portuguese vineyards soils: suggesting a ‘Galleria-Tenebrio-bait method’ as bait-insects Galleria and Tenebrio significantly underestimate the respective recoveries of Metarhizium (robertsii) and Beauveria (bassiana). Mycokeys, 38, 1-23. https://doi.org/10.3897/mycokeys.38.26970
Thao, L. D., Anh, P. T., Trang, T. T. T., Khanh, T. N., Hien, L. T., Binh, V. T. P., Thanh, H. M., & Thanh, D. T. (2021). Fusarium falciforme, a pathogen causing wilt disease of chrysanthemum in Vietnam. New Disease Report, 43(2), e12013. https://doi.org/10.1002/ndr2.12013
Thao, P. T. P., Toan, L. T., Yen, L. T. H., and Cam, N. T. M. (2023). Effect of two growing media types and times of pinching on growth, flower quality of purple Pico (Chrysanthemum sp.). Vietnam Journal of Agriculture & Rural Development, 20/2023, 3-11.
Thrane, U. (1999). Fusarium. In: Thrane, U., Robinson, R., Batt, C., Patel, P. (Eds.), Encyclopedia of food microbiology (pp. 901-906). Academic Press.
Wang, X., Wang, Z., Yu, J., Wu, L., Zhang, X., Luo, F., Chen, Z., & Zhou, L. (2023). Method validation for multi-pesticide residue determination in chrysanthemum. Molecules (Basel, Switzerland), 28(3), 1291. https://doi.org/10.3390/molecules28031291
Zalila-Kolsi, I., Mahmoud, A. B., Ali, H., Sellami, S., Nasfi, Z., Tounsi, S., & Jamoussi, K. (2016). Antagonist effects of Bacillus spp. strains against Fusarium graminearum for protection of durum wheat (Triticum turgidum L. subsp. durum). Microbiology Research, 192, 148–158. https://doi.org/10.1016/j.micres.2016.06.012
Zubrod, J. P., Bundschuh, M., Arts, G., Brühl, C. A., Imfeld, G., Knäbel, A., Payraudeau, S., Rasmussen, J. J., Rohr, J., Scharmüller, A., Smalling, K., Stehle, S., Schulz, R., & Schäfer, R. B. (2019). Fungicides: an overlooked pesticide class?. Environmental Science and Technology, 53(7), 3347–3365. https://doi.org/10.1021/acs.est.8b04392