title: MaMk1, a FUS3/KSS1-type mitogen-activated protein kinase gene, is required for appressorium formation, and insect cuticle penetration of the entomopathogenic fungus Metarhizium acridum.
abstract: Entomopathogenic fungi have great potential for development as insecticides. However, large-scale use of mycoinsecticides is partially limited by poor efficiency. In many fungal pathogens, the yeast and fungal extracellular signal-regulated kinase (YERK1) subfamily is crucial to the fungal pathogenicity. In this study, a Fus3/Kss1-type mitogen-activated protein kinase (MAPK) gene MaMk1 (GenBank accession No. EFY93607) was identified in Metarhizium acridum, which encodes a member of the YERK1 subfamily. Targeted gene disruption was used to analyze the function of MaMk1 in fungal growth, conidial yield and virulence. Growth assays showed that MaMk1 disruption did not affect fungal growth and conidial yield on potato dextrose agar (PDA) plates. Bioassays by topical inoculation showed that a MaMk1-disruption mutant entirely lost its pathogenicity for the locusts, likely because of failure to penetrate the insect cuticle, which might have been caused by inability to form appressoria during infection. However, bioassays by injection showed no significant difference in virulence among the wild type (WT), ΔMaMk1 mutant and complementary transformant. ΔMaMk1 mutant failed to penetrate the cuticle outwards and sporulate on the locust cadaver. These results suggest that MaMk1 is required for penetration of the insect cuticle both into the hemocele and outside from the hemocele, but is dispensable for fungal growth in insect hemolymph. Gene expression pattern analysis showed that MaMk1 disruption downregulated expression of Mad1 and Mpl1, but did not reduce expression of Pr1 in M. acridum.
authors.affiliation: Genetic Engineering Research Center, School of Life Sciences, Chongqing University, Chongqing 400030, PR China; Chongqing Engineering Research Center for Fungal Insecticide, Chongqing 400030, PR China; Key Laboratory of Gene Function and Regulation Technologies under Chongqing Municipal Education Commission, Chongqing 400030, PR China.
authors.forename: Kai
authors.initials: K
authors.lastname: Jin
authors.affiliation: Research and Development Center of Biorational Pesticide, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Lirong
authors.initials: L
authors.lastname: Han
authors.affiliation: Genetic Engineering Research Center, School of Life Sciences, Chongqing University, Chongqing 400030, PR China; Chongqing Engineering Research Center for Fungal Insecticide, Chongqing 400030, PR China; Key Laboratory of Gene Function and Regulation Technologies under Chongqing Municipal Education Commission, Chongqing 400030, PR China. Electronic address: yuxianxia@cqu.edu.cn.
authors.forename: Yuxian
authors.initials: Y
authors.lastname: Xia
mesh_terms: D000818:Animals; D001483:Base Sequence; D005656:Fungal Proteins; D005800:Genes, Fungal; D006110:Grasshoppers / Q000469:parasitology*; D052981:Metarhizium / Q000235:genetics* / Q000472:pathogenicity*; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics*; D008969:Molecular Sequence Data; D010572:Pest Control, Biological / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D014774:Virulence
chemical_list: D005656:Fungal Proteins; D020928:Mitogen-Activated Protein Kinases
keywords: Appressorium formation; Fus3/Kss1-type MAPK; MaMk1; Metarhizium acridum; Penetration; Virulence