title: Characterization of the sterol 14α-demethylases of Fusarium graminearum identifies a novel genus-specific CYP51 function.
abstract: CYP51 encodes the cytochrome P450 sterol 14α-demethylase, an enzyme essential for sterol biosynthesis and the target of azole fungicides. In Fusarium species, including pathogens of humans and plants, three CYP51 paralogues have been identified with one unique to the genus. Currently, the functions of these three genes and the rationale for their conservation within the genus Fusarium are unknown. Three Fusarium graminearum CYP51s (FgCYP51s) were heterologously expressed in Saccharomyces cerevisiae. Single and double FgCYP51 deletion mutants were generated and the functions of the FgCYP51s were characterized in vitro and in planta. FgCYP51A and FgCYP51B can complement yeast CYP51 function, whereas FgCYP51C cannot. FgCYP51A deletion increases the sensitivity of F. graminearum to the tested azoles. In ΔFgCYP51B and ΔFgCYP51BC mutants, ascospore formation is blocked, and eburicol and two additional 14-methylated sterols accumulate. FgCYP51C deletion reduces virulence on host wheat ears. FgCYP51B encodes the enzyme primarily responsible for sterol 14α-demethylation, and plays an essential role in ascospore formation. FgCYP51A encodes an additional sterol 14α-demethylase, induced on ergosterol depletion and responsible for the intrinsic variation in azole sensitivity. FgCYP51C does not encode a sterol 14α-demethylase, but is required for full virulence on host wheat ears. This is the first example of the functional diversification of a fungal CYP51.
authors: Department of Plant Pathology, China Agricultural University, Beijing, 100193, China.|Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Jieru
authors: J
authors: Fan
authors: Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Martin
authors: M
authors: Urban
authors: Institute of Life Science and College of Medicine, Swansea University, Swansea, SA2 8PP, UK.
authors: Josie E
authors: JE
authors: Parker
authors: Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Helen C
authors: HC
authors: Brewer
authors: Institute of Life Science and College of Medicine, Swansea University, Swansea, SA2 8PP, UK.
authors: Steven L
authors: SL
authors: Kelly
authors: Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Kim E
authors: KE
authors: Hammond-Kosack
authors: Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Bart A
authors: BA
authors: Fraaije
authors: Department of Plant Pathology, China Agricultural University, Beijing, 100193, China.
authors: Xili
authors: X
authors: Liu
authors: Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, AL5 2JQ, UK.
authors: Hans J
authors: HJ
authors: Cools
mesh_terms: D000577:Amides / Q000494:pharmacology; D017360:Arabidopsis / Q000382:microbiology; D001393:Azoles / Q000494:pharmacology; D025141:Drug Resistance, Fungal / Q000187:drug effects / Q000235:genetics; D004875:Ergosterol / Q000235:genetics / Q000378:metabolism; D019143:Evolution, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005670:Fusarium / Q000187:drug effects* / Q000201:enzymology* / Q000472:pathogenicity* / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D007810:Lanosterol / Q000031:analogs & derivatives / Q000378:metabolism; D027845:Malus / Q000382:microbiology; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013172:Spores, Fungal / Q000235:genetics; D058886:Sterol 14-Demethylase / Q000235:genetics / Q000378:metabolism*; D014255:Trichothecenes / Q000378:metabolism; D014908:Triticum / Q000382:microbiology; D014774:Virulence / Q000235:genetics
chemical_list: D000577:Amides; D001393:Azoles; D005656:Fungal Proteins; D014255:Trichothecenes; D007810:Lanosterol; C014520:ebericol; D058886:Sterol 14-Demethylase; C007262:deoxynivalenol; C451426:N-(2,3-dichloro-4-hydroxyphenyl)-1-methylcyclohexanecarboxamide; D004875:Ergosterol