title: DNA methyltransferases contribute to the fungal development, stress tolerance and virulence of the entomopathogenic fungus Metarhizium robertsii.
abstract: DNA methylation is an important epigenetic mark in mammals, plants, and fungi and depends on multiple genetic pathways involving de novo and maintenance DNA methyltransferases (DNMTases). Metarhizium robertsii, a model system for investigating insect-fungus interactions, has been used as an environmentally friendly alternative to chemical insecticides. However, little is known concerning the molecular basis for DNA methylation. Here, we report on the roles of two DNMTases (MrRID and MrDIM-2) by characterizing ΔMrRID, ΔMrDIM-2, and ΔRID/ΔDIM-2 mutants. The results showed that approximately 71, 10, and 8% ofmC sites remained in the ΔMrRID, ΔMrDIM-2, and ΔRID/ΔDIM-2 strains, respectively, compared with the wild-type (WT) strain. Further analysis showed that MrRID regulates the specificity of DNA methylation and MrDIM-2 is responsible for most DNA methylation, implying an interaction or cooperation between MrRID and MrDIM-2 for DNA methylation. Moreover, the ΔMrDIM-2 and ΔRID/ΔDIM-2 strains showed more defects in radial growth and conidial production compared to the WT. Under ultraviolet (UV) irradiation or heat stress, an obvious reduction in spore viability was observed for all the mutant strains compared to the WT. The spore median lethal times (LT50s) for the ΔMrDIM-2 and ΔRID/ΔDIM-2 strains in the greater wax moth, Galleria mellonella, were decreased by 47.7 and 65.9%, respectively, which showed that MrDIM-2 is required for full fungal virulence. Our data advances the understanding of the function of DNMTase in entomopathogenic fungi, which should contribute to future epigenetic investigations in fungi.
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Wang
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Lintao
authors.initials: L
authors.lastname: Qiao
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Jianyu
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Jinrui
authors.initials: J
authors.lastname: Fan
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.|School of Plant Protection, Anhui Agricultural University, Hefei, 230036, China.
authors.forename: Zhang-Xun
authors.initials: ZX
authors.lastname: Wang
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.|Engineering Technology Research Center of Microbial Fermentation Anhui Province, Anhui Polytechnic University, Wuhu, 241000, China.
authors.forename: Wanzhen
authors.initials: W
authors.lastname: Li
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China.|Department of Food and Biology, Xuzhou Institute of Technology, Xuzhou, 221008, China.
authors.forename: Anhui
authors.initials: A
authors.lastname: Chen
authors.affiliation: Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, 230036, China. bhuang@ahau.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Huang
mesh_terms: D000818:Animals; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D018869:Heat-Shock Response / Q000235:genetics; D006358:Hot Temperature; D007313:Insecta / Q000382:microbiology; D052981:Metarhizium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D009036:Moths / Q000382:microbiology; D010641:Phenotype; D013172:Spores, Fungal; D013312:Stress, Physiological; D014466:Ultraviolet Rays; D014774:Virulence
chemical_list: D015254:DNA Modification Methylases
keywords: DNA methylation; DNA methyltransferase; Metarhizium robertsii; Phenotype; Virulence