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Identification and Functional Characterization of the Heat Shock Protein (HSP) 40 Encoding Gene, MoMHF6, in Magnaporthe oryzae
Received date: 2023-03-30
Revised date: 2023-04-20
Online published: 2023-11-14
【Objective】 To explore the roles of chaperone HSP40 (heat shock protein 40) in morphological differentiation and pathogenesis in Magnaporthe oryzae. 【Method】 The HSP40 encoding gene MoMHF6 of M. oryzae was knocked out by DNA homologous recombination, and a deletion mutant ΔMomhf6 was generated. Biological function of the gene was systematically characterized via phenotypic analysis, gene complementation and RNA-seq analysis. 【Result】 Phenotypic analysis of the mutant revealed severe impairments in aerial hypha growth, asexual sporulation, conidial germination, perithecium production and appressorium formation. However, compared to the wild-type strain, the deletion of MoMHF6 did not affect mycelium radial growth on CM medium and the development of asci and ascospores in perithecia. Incipient cytorrhysis assays revealed that the appressorium collapse rate of the ΔMomhf6 mutant under treatment with 1, 2 or 3 mol/L glycerol was significantly increased compared to the wild-type strain, indicating that MoMHF6 is required for appressorium turgor generation. Also, we found that the ΔMomhf6 mutant was unable to penetrate into onion epidermis and completely nonpathogenic to susceptible hosts. Even in wounded barley leaves, infectious growth of the mutant was severely impaired. Moreover, the deletion of MoMHF6 resulted in significantly increased sensitivity to oxidative stress and delayed glycogen transportation and degradation during appressorium development, indicating that MoMHF6 is involved in oxidative stress response and glycogen metabolism of appressorium. All phenotypic defects of the ΔMomhf6 mutant could be restored by reintroducing the full-length MoMHF6 gene into the mutant. In addition, RNA-seq analysis of the ΔMomhf6 mutant revealed that several known pathogenicity-related genes in differentially expressed genes (DEGs) might be regulated by MoMHF6, such as MoATG4, MoPL1, MoVPR, and GAS1. 【Conclusion】 The HSP40 gene MoMHF6 plays an important role in asexual sporulation, appressorium formation, host penetration, oxidative stress response and pathogenicity in M. oryzae. These results are of great significance for further understanding the gene networks and molecular mechanisms in regulating morphological differentiation and pathogenesis governed by MoMHF6 in M. oryzae.
Key words： Magnaporthe oryzae; MoMHF6; heat shock protein 40; functional analysis; RNA-seq
TONG Qi, WANG Chunyan, QUE Yawei, XIAO Yu, WANG Zhengyi . Identification and Functional Characterization of the Heat Shock Protein (HSP) 40 Encoding Gene, MoMHF6, in Magnaporthe oryzae[J]. Chinese Journal OF Rice Science, 2023 , 37(6) : 563 -576 . DOI: 10.16819/j.1001-7216.2023.230301
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