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| Identification and Functional Analysis of the Secretory Protein Feruloyl Esterase (FpESD) in Fusarium proliferatum |
| WANG Yi-Tong, Li Hai-Bo, WANG Yu-Qing, WU Zhi-Hong, GAO Yi-Zhou* |
| School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China |
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Abstract Fusarium proliferatum is a phytopathogenic fungus with a wide host range and serious damage, but its pathogenic mechanism is not well understood. Effectors play an important role in the infection process of phytopathogenic fungi. The prediction, screening, and identification of effectors are of great importance in exploring the mechanism of host-pathogen interaction. In this study, based on the transcriptome sequencing results of F. proliferatum, 43 putative effectors were screened by signal peptide identification, transmembrane domain screening, cysteine number analysis, and Pathogen Host Interactions database (PHI-base) analysis. Functional studies were carried out on one of the candidate effectors, feruloyl esterase FpESD. Yeast system validation revealed that the signal peptide encoded by the N-terminal 20 amino acids of FpESD possesses secretion activity. The homologous recombination strategy was used to generate the FpESD knockout mutant (ΔFpESD), and the complementary strain (ΔFpESD-C) was obtained through complementation experiments. Growth rate measurements, spore yield assessments, and spore length analysis demonstrated that the deletion of the FpESD gene significantly reduced fungal growth rate and conidia production (P<0.05), with shorter conidia length observed for the knockout mutant. Additionally, environmental stress tests revealed that ΔFpESD exhibited significantly reduced sensitivity to CaCl2, MgCl2, and Congo red (P<0.05), indicating that FpESD was involved in regulating the response to environmental stress. Pathogenicity assays using plate inoculation showed that the disease index of alfalfa (Medicago sativa) seedlings infected by ΔFpESD was significantly lower than that of the wild-type strain (P<0.05), suggesting that FpESD positively regulated the pathogenicity of F.proliferatum. This study helps to elucidate the pathogenic molecular mechanism of F. proliferatum, and also provides ideas for the in-depth study of F. proliferatum effectors.
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Received: 23 April 2024
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Corresponding Authors:
* gaoyz@zju.edu.cn
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