Abstract
Objectives
Microsclerotia (MS), anti-stress structures produced by many filamentous fungi, have been proven to be a great substitute for conidia in the production of insecticides within entomogenous fungi. NADPH oxidase (Nox) is a highly conserved ROS-response protein family that is widespread in eukaryotes and plays distinct roles in environmental fitness among various filamentous fungi. However, it is not clear whether the formation of MS and pathogenicity in entomogenous fungi is regulated by the Nox inside. In this study, we reported the presence of NADPH oxidase homologs in a great potential biocontrol fungus, Metarhizium rileyi, and further showed multiple biological functions.
Results
Three Nox homologous genes in M. rileyi showed high expression throughout the entire process of MS formation. Targeted deletion of MrNoxA, MrNoxB and MrNoxR all led to a decrease in MS yield and impaired morphology. Moreover, the anti-adversity assay showed that they are indispensable for growth, osmotic pressure and oxidative stress regulation in Metarhizium rileyi. Most importantly, △MrNoxR and △MrNoxA but not △MrNoxB showed a dramatic reduction in virulence via inoculation. The normality of appressoria might be unaffected in mutants since there are no striking differences in virulence compared with WT by topical injections.
Conclusion
Our results revealed that NADPH oxidase plays important roles in growth regulation, MS formation and pathogenicity in M. rileyi, perhaps in the ROS response and hyphal polarity.
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Funding
This work was supported by Department of Science and Technology Planning Project of Henan Province (Nos. 222102110102, 222102110015, 212102110142) and Scientific Research Support Project of Zhoukou Normal University (ZKNUC2021049, ZKNUC2021050 and ZKNUC2020045).
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YL and LF designed the experiments. BL, XL, JW and FM conducted the experiments under the supervision of YL and LF. LF, FD and HL wrote and revised the manuscript.
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Fan, L., Li, B., Wang, J. et al. Multifunctional regulation of NADPH oxidase in growth, microsclerotia formation and virulence in Metarhizium rileyi. Biotechnol Lett 45, 1441–1455 (2023). https://doi.org/10.1007/s10529-023-03427-2
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DOI: https://doi.org/10.1007/s10529-023-03427-2