Plant Protection (Scientific Journal of Agriculture)

Plant Protection (Scientific Journal of Agriculture)

Analysis of phytohormone-related gene expression in cotton following Beauveria bassiana GHA endophytic association

Document Type : Research paper-English

Authors
Department of Plant Protection, Ferdowsi University of Mashhad, Mashhad, Iran
Abstract
Cotton (Gossypium hirsutum L.) is a vital global crop but suffers heavy losses from insect pests that reduce yield and fiber quality. Conventional pesticide-based control causes environmental harm, underscoring the need for sustainable alternatives. Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales), an entomopathogenic fungus, offers promise as both a biocontrol agent and a plant endophyte that can enhance host defense responses. This study investigates how inoculation with live or heat-killed spores of B. bassiana GHA affects the expression of defense-related genes in cotton, focusing on key components of the ethylene, salicylic acid, and jasmonic acid signaling pathways. Four genes ERF1 and MPK2 (ethylene pathway), WRKY (salicylic acid pathway), and JAZ1 (jasmonic acid pathway) were analyzed at 2, 8, 24, and, 48 hours post-inoculation. Results showed that live spores markedly upregulated all target genes compared with controls, indicating strong systemic activation of defense signaling. ERF1 and MPK2 expression peaked at 48 hours, showing 46.55-fold and 29.75-fold increases, while WRKY and JAZ1 peaked at 24 hours with 26 and 21.98-fold increases, respectively. Heat-killed B. bassiana spores caused a significant increase in ERF1 and MPK2 expression (12.86- and 12.11-fold at 2–8 hours), while JAZ and WRKY genes showed no significant change. These findings demonstrate that B. bassiana can induce broad-spectrum systemic resistance in cotton through coordinated activation of multiple hormonal pathways. The results highlight its potential to strengthen plant immunity and reduce dependence on chemical pesticides, offering a promising strategy for sustainable pest management and environmentally resilient cotton production.
Keywords
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Bamisile, B. S., Dash, C. K., Akutse, K. S., Keppanan, R., & Wang, L. (2018). Fungal endophytes: beyond herbivore management. Frontiers in Microbiology, 9, 1-11. https://doi.org/10.3389/fmicb.2018.00544
Bamisile, B. S., Siddiqui, J. A., Akutse, K. S., Aguila, L. C. R., & Xu, Y. (2021). General Limitations to Endophytic Entomopathogenic Fungi Use as Plant Growth Promoters, Pests and Pathogens Biocontrol Agents. Plants, 10(10).
Cavazos-Vallejo, T., Valadez-Lira, J. A., Orozco-Flores, A. A., Gomez-Flores, R., Ek-Ramos, M. J., Quistián-Martínez, D., Alcocer-González, J. M., & Tamez-Guerra, P. (2023). In Planta Detection of Beauveria bassiana (Ascomycota: Hypocreales) Strains as Endophytes in Bean (Phaseolus vulgaris L.). Plants, 13(1). https://doi.org/10.3390/plants13010022
Chi, B.-j., Zhang, D.-m., & Dong, H.-z. (2021). Control of cotton pests and diseases by intercropping: A review. Journal of Integrative Agriculture, 20(12), 3089-3100.
Christian, N., Sedio, B. E., Florez-Buitrago, X., Ramírez-Camejo, L. A., Rojas, E. I., Mejía, L. C., Palmedo, S., Rose, A., Schroeder, J. W., & Herre, E. A. (2020). Host affinity of endophytic fungi and the potential for reciprocal interactions involving host secondary chemistry. American Journal of Botany, 107(2), 219–228. https://doi.org/10.1002/ajb2.1436
Dannon, H. F., Dannon, A. E., Douro-Kpindou, O. K., Zinsou, A. V., Houndete, A. T., Toffa-Mehinto, J., Elegbede, I. M., Olou, B. D., & Tamò, M. (2020). Toward the efficient use of Beauveria bassiana in integrated cotton insect pest management. Journal of Cotton Research 3(24), 1-21. https://doi.org/10.1186/s42397-020-00061-5
Donga, T. K., Vega, F. E., & Klingen, I. (2018). Establishment of the fungal entomopathogen Beauveria bassiana as an endophyte in sugarcane, Saccharum officinarum. Fungal Ecology, 35, 70-77. https://doi.org/10.1016/j.funeco.2018.06.008
Ebrahimifar, J., Jamshidnia, A., & Sadeghi, R. (2017). Pathogenicity of Beauveria bassiana and Metarhizium anisopliae on elm leaf beetle, Xanthogaleruca luteola Muller (Col., Chrysomelidae). Plant Protection (Scientific Journal of Agriculture), 40(2), 23-32. https://doi.org/10.22055/PPR.2017.12833
Farrokhzadeh, H., Jaronski, S. T., & Rashed, A. (2025). Reduced survivorship, host preference, and feeding damage by Helicoverpa zea (Lepidoptera: Noctuidae) on cotton plants colonized by the endophyte Beauveria bassiana (Ascomycota: Hypocreales). Journal of Economic Entomology, 118(2), 523–530. https://doi.org/10.1093/jee/toae302
Farrokhzadeh, H., Sharifi, S., Eroğlu, G. B., & Karimi, J. (2024). A new fungal entomopathogen has potency as a biocontrol agent of longhorn beetle larva, Osphranteria coerulescencs. International Journal of Tropical Insect Science, 44(3), 1185 - 1193. https://doi.org/10.1007/s42690-024-01230-8
Fesel, P. H., & Zuccaro, A. (2016). β-glucan: Crucial component of the fungal cell wall and elusive MAMP in plants. Fungal Genetics and Biology, 90, 53-60. https://doi.org/10.1016/j.fgb.2015.12.004
Goettel, M. S., & Inglis, G. D. (1997). Fungi: Hyphomycetes. In L. A. Lacey (Ed.), Manial of Techniques in Insect Pathology (pp. 213–247). Academic Press. https://doi.org/10.1016/B978-012432555-5/50013-0
Hagenbucher, S., Olson, D., Ruberson, J. R., Wäckers, F., & Romeis, J. (2013). Resistance Mechanisms Against Arthropod Herbivores in Cotton and Their Interactions with Natural Enemies. Critical Reviews in Plant Sciences, 32(6), 458–482. https://doi.org/10.1080/07352689.2013.809293
Hashemi, L., Golparvar, A. R., Nasr-Esfahani, M., & Golabadi, M. (2020). Expression analysis of defense-related genes in cucumber (Cucumis sativus L.) against Phytophthora melonis. Molecular Biology Reports, 47(7), 4933-4944. https://doi.org/10.1007/s11033-020-05520-5
IBM-SPSS. (2024). IBM SPSS Statistics for Windows. In (Version 29) IBM Corp.
Iida, Y., Higashi, Y., Nishi, O., Kouda, M., Maeda, K., Yoshida, K., Asano, S., Kawakami, T., Nakajima, K., Kuroda, K., Tanaka, C., Sasaki, A., Kamiya, K., Yamagishi, N., Fujinaga, M., Terami, F., Yamanaka, S., & Kubota, M. (2023). Entomopathogenic fungus Beauveria bassiana–based bioinsecticide suppresses severity of powdery mildews of vegetables by inducing the plant defense responses. Frontiers in Plant Science, 14, 12. https://doi.org/10.3389/fpls.2023.1211825
Jaronski, S. T. (2010). Ecological factors in the inundative use of fungal entomopathogens. BioControl, 55, 159-185. https://doi.org/10.1007/s10526-009-9248-3
Jaronski, S. T., & Jackson, M. A. (2012). Mass production of entomopathogenic Hypocreales. In L. L.A (Ed.), Manual of Techniques in Invertebrate Pathology (Vol. 2, pp. 255-284). Elsevier/Academic Press. https://doi.org/10.1016/B978-0-12-386899-2.00008-7
Karooei, P., Darvishnia, M., Bazgir, E., & Mirzaeipour, Z. (2025). Investigating effect of the endophytic of two Beauveria species on growth traits and some biochemical activities of bean (Phaseolus vulgaris). Plant Protection (Scientific Journal of Agriculture), 47(4), 1-21. https://doi.org/10.22055/PPR.2024.47925.1760
Koornneef, A., & Pieterse, C. M. (2008). Cross talk in defense signaling. Plant Physiology, 146(3), 839–844. https://doi.org/10.1104/pp.107.112029
Li, J., Zhu, L., Hull, J. J., Liang, S., Daniell, H., Jin, S., & Zhang, X. (2016). Transcriptome analysis reveals a comprehensive insect resistance response mechanism in cotton to infestation by the phloem feeding insect Bemisia tabaci (whitefly). Plant Biotechnology Journal, 14(10), 1956–1975.
Li, N., Han, X., Feng, D., Yuan, D., & Huang, L.-J. (2019). Signaling crosstalk between salicylic acid and ethylene/jasmonate in plant defense: do we understand what they are whispering? International Journal of Molecular Sciences, 20(3). https://doi.org/10.3390/ijms20030671
Liu, L., Liu, S., Meng, Q., Chen, B., Zhang, J., Zhang, X., Lin, Z., & Zou, Z. (2025). Evaluating Beauveria bassiana Strains for Insect Pest Control and Endophytic Colonization in Wheat. Insects, 16(3). https://doi.org/10.3390/insects16030287
Lorenzo, O., Piqueras, R., Sánchez-Serrano, J. J., & Solano, R. (2003). Ethylene response factor1 itegrates signals from ethylene and jasmonate pthways in plant defense. Plant Cell, 15(1), 165–178. https://doi.org/10.1105/tpc.007468
Mascarin, G. M., & Jaronski, S. T. (2016). The production and uses of Beauveria bassiana as a microbial insecticide. World Journal of Microbiology and Biotechnology, 32(11), 177. https://doi.org/10.1007/s11274-016-2131-3
Mehrmoradi, H., Jamali, S., & Pourian, H. R. (2022). Isolation of entomopathogenic fungi from cultivated and uncultivated soils and evaluation of virulence against Callosobruchus maculatus (F.) (Coleoptera:Chrysomelidae). Journal of Entomological Society of Iran, 42(1), 29-53.  https://doi.org/10.52547/JESI.42.1.3
Naseri, M., Bemani, M., Alipanah, H., Noorbakhsh, S., & Zohdi, H. (2023). First report of the fall armyworm, Spodoptera frugiperda (Lepidoptera, Noctuidae: Noctuinae) from Iran. Journal of Entomological Society of Iran 44(1). https://doi.org/10.61186/JESI.44.1.9
Nishi, O., Sushida, H., Higashi, Y., & Iida, Y. (2020). Epiphytic and endophytic colonisation of tomato plants by the entomopathogenic fungus Beauveria bassiana strain GHA. Mycology, 12(1), 39–47. https://doi.org/10.1080/21501203.2019.1707723
Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research, 29(9), e45. https://doi.org/10.1093/nar/29.9.e45
Qin, X., Zhao, X., Huang, S., Deng, J., Li, X., Luo, Z., & Zhang, Y. (2021). Pest management via endophytic colonization of tobacco seedlings by the insect fungal pathogen Beauveria bassiana. Pest Management Science, 77(4), 2007-2018. https://doi.org/10.1002/ps.6229
Quesada-Moraga, Munoz-Ledesma, & Santiago-Alvarez. (2009). Systemic protection of Papaver somniferum L. against Iraella luteipes (Hymenoptera: Cynipidae) by an endophytic strain of Beauveria bassiana (Ascomycota: Hypocreales). Environmental Entomology, 38, 723–730. https://doi.org/10.1603/022.038.0324
Raad, M., Glare, T. R., Brochero, H. L., Müller, C., & Rostás, M. (2019). Transcriptional reprogramming of Arabidopsis thaliana defence pathways by the entomopathogen Beauveria bassiana correlates with resistance against a fungal pathogen but not against insects. Frontier In Microbiology, 10. https://doi.org/10.3389/fmicb.2019.00615
Rajendran, T. P., Birah, A., & Burange, P. S. (2018). Insect Pests of Cotton. In Omkar (Ed.), Pests and Their Management. Springer Nature. https://doi.org/10.1007/978-981-10-8687-8_11
Silva, A. C. L., Silva, G. A., Abib, P. H. N., Carolino, A. T., & Samuels, R. I. (2020). Endophytic colonization of tomato plants by the entomopathogenic fungus Beauveria bassiana for controlling the south american tomato pinworm, Tuta absoluta. CABI Agriculture and Bioscience, 3(1). https://doi.org/10.1186/s43170-020-00002-x
Tall, S., & Meyling, N. V. (2018). Probiotics for Plants? Growth Promotion by the Entomopathogenic Fungus Beauveria bassiana Depends on Nutrient Availability. Microbial Ecology, 76, 1002–1008. https://doi.org/10.1007/s00248-018-1180-6
Wang, L., Guo, D., Zhao, G., Wang, J., Zhang, S., Wang, C., & Guo, X. (2022). Group IIc WRKY transcription factors regulate cotton resistance to Fusarium oxysporum by promoting GhMKK2-mediated flavonoid biosynthesis New Phytologist, 326(1), 249-265. https://doi.org/10.1111/nph.18329
Wang, M., Wang, Q., & Zhang, B. (2013). Evaluation and selection of reliable reference genes for gene expression under abiotic stress in cotton (Gossypium hirsutum L.). Gene, 530(1), 44-50. https://doi.org/10.1016/j.gene.2013.07.084
Yan, L., Zhu, J., Zhao, X., Shi, J., Jiang, C., & Shao, D. (2019). Beneficial effects of endophytic fungi colonization on plants. Applied Microbiology and Biotechnology, 103(2), 3327–3340. https://doi.org/10.1007/s00253-019-09713-2
Zamani, S. M. , Alinejad, M. , Askary, H. , Gholami Gavamabad, R. , Sepasi, N. , Farahani, S. and Ahangaran, Y. (2025). Field performance of trunk-applied entomopathogenic fungi for management of the oak moth, Leucoma wiltshirei (Lepidoptera: Erebidae) during a two-year study. Journal of Entomological Society of Iran, 45(4), 663-678. doi: https://doi.org/10.22034/jesi.45.4.14
Zhang, P., Jackson, E., Li, X., & Zhang, Y. (2025). Salicylic acid and jasmonic acid in plant immunity. Horticulture Research, 12(7). https://doi.org/10.1093/hr/uhaf082
Zhang, Z., Wang, P., Luo, X., Yang, C., Tang, Y., Wang, Z., Hu, G., Ge, X., Xia, G., & Wu, J. (2019). Cotton plant defence against a fungal pathogen is enhanced by expanding BLADE-ON-PETIOLE1 expression beyond lateral-organ boundaries. Communications Biology, 238.
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