Abstract
Mitogen-activated protein kinase (MAPK) cascades are broadly conserved and play essential roles in multiple cellular processes, including fungal development, pathogenicity, and secondary metabolism. Their function, however, also exhibits species and strain specificity. Penicillium oxalicum secretes plant-biomass-degrading enzymes (PBDEs) that contribute to the carbon cycle in the natural environment and to utilization of lignocellulose in industrial processes. However, knowledge of the MAPK pathway in P. oxalicum has been relatively limited. In this study, comparative transcriptomic analysis of P. oxalicum, cultured on different carbon sources, found ten putative kinase genes with significantly modified transcriptional levels. Six of these putative kinase genes were knocked out in the parental strain ∆PoxKu70, and deletion of the gene, Fus3/Kss1-like PoxMK1 (POX00158), resulted in the largest reduction (91.1%) in filter paper cellulase production. Further tests revealed that the mutant ∆PoxMK1 lost 37.1 to 92.2% of PBDE production, under both submerged- and solid-state fermentation conditions, compared with ∆PoxKu70. In addition, the mutant ∆PoxMK1 had reduced vegetative growth and increased pigment biosynthesis. Comparative transcriptomic analysis showed that PoxMK1 deletion from P. oxalicum downregulated the expression of major PBDE genes and known regulatory genes such as PoxClrB and PoxCxrB, whereas the transcription of pigment biosynthesis-related genes was upregulated. Comparative phosphoproteomic analysis revealed that PoxMK1 deletion considerably modified phosphorylation of key transcription- and signal transduction-associated proteins, including transcription factors Mcm1 and Atf1, RNA polymerase II subunits Rpb1 and Rpb9, MAPK-associated Hog1 and Ste7, and cyclin-dependent kinase Kin28. These findings provide novel insights into understanding signal transduction and regulation of PBDE gene expression in fungi.
Key points
• PoxMK1 is involved in expression of PBDE- and pigment synthesis-related genes.
• PoxMK1 is required for vegetative growth of P. oxalicum.
• PoxMK1 is involved in phosphorylation of key TFs, kinases, and RNA polymerase II.
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Funding
This work was financially supported by the National Natural Science Foundation of China (grants 31760023 and 31660305), the Autonomous Research Project of State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources (SKLCUSA-a201902 and SKLCUSA-a201923), the Training Program for 1000 Young and Middle-aged Key Teachers in Guangxi at 2019, and the One Hundred Person Project of Guangxi.
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JXF supervised this study and revised the manuscript. SZ co-supervised and directed all experiments and revised the manuscript. BM conducted construction of deletion mutants, measurement of enzymatic activities, transcriptomes, and phosphoproteomes, and wrote the manuscript. YNN performed construction of complementary strain and enzymatic activity assay. CXL conducted bioinformatic analysis. DT and HG conducted RT-qPCR analysis. XMP performed phenotypic analyses. XML was involved in preparation of experimental materials.
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Ma, B., Ning, YN., Li, CX. et al. A mitogen-activated protein kinase PoxMK1 mediates regulation of the production of plant-biomass-degrading enzymes, vegetative growth, and pigment biosynthesis in Penicillium oxalicum. Appl Microbiol Biotechnol 105, 661–678 (2021). https://doi.org/10.1007/s00253-020-11020-0
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DOI: https://doi.org/10.1007/s00253-020-11020-0