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Fungal pathogens of plants and insects infect their hosts by direct penetration of the cuticle. Plant and insect cuticles are covered by a hydrocarbon-rich waxy outer layer that represents the first barrier against infection. However, the fungal genes that underlie insect waxy layer degradation have received little attention. Here we characterize the single cytochrome P450 monoxygenase family 52 ( MrCYP52 ) gene of the insect pathogen  Metarhizium robertsii , and demonstrate that it encodes an enzyme required for efficient utilization of host hydrocarbons. Expressing a green florescent protein gene under control of the  MrCYP52  promoter confirmed that  MrCYP52  is up regulated on insect cuticle as well as by artificial media containing decane (C10), extracted cuticle hydrocarbons, and to a lesser extent long chain alkanes. Disrupting  MrCYP52  resulted in reduced growth on epicuticular hydrocarbons and delayed developmental processes on insect cuticle, including germination and production of appressoria (infection structures). Extraction of alkanes from cuticle prevented induction of  MrCYP52  and reduced growth. Insect bioassays against caterpillars ( Galleria mellonella ) confirmed that disruption of  MrCYP52  significantly reduces virulence. However,  MrCYP52  was dispensable for normal germination and appressorial formation  in vitro  when the fungus was supplied with nitrogenous nutrients. We conclude therefore that  MrCYP52  mediates degradation of epicuticular hydrocarbons and these are an important nutrient source, but not a source of chemical signals that trigger infection processes.

The MrCYP52 Cytochrome P450 Monoxygenase Gene of Metarhizium robertsii Is Important for Utilizing Insect Epicuticular Hydrocarbons