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The endophytic insect pathogenic fungi (EIPF)  Metarhizium  promotes plant growth through symbiotic association and the transfer of insect-derived nitrogen. However, little is known about the genes involved in this association and the transfer of nitrogen. In this study, we assessed the involvement of six  Metarhizium robertsii  genes in endophytic, rhizoplane and rhizospheric colonization with barley roots. Two ammonium permeases ( MepC  and  Mep2)  and a  urease , were selected since homologous genes in arbuscular mycorrhizal fungi were reported to play a pivotal role in nitrogen mobilization during plant root colonization. Three other genes were selected on the basis on RNA-Seq data that showed high expression levels on bean roots, and these encoded a hydrophobin (Hyd3), a subtilisin-like serine protease (Pr1A) and a hypothetical protein. The root colonization assays revealed that the deletion of  urease ,  hydrophobin ,  subtilisin-like serine protease  and  hypothetical protein  genes had no impact on endophytic, rhizoplane and rhizospheric colonization at 10 or 20 days. However, the deletion of  MepC  resulted in significantly increased rhizoplane colonization at 10 days whereas Δ Mep2  showed increased rhizoplane colonization at 20 days. In addition, the nitrogen transporter mutants also showed significantly higher  15 N incorporation of insect derived nitrogen in barley leaves in the presence of nutrients. Insect pathogenesis assay revealed that disruption of  MepC ,  Mep2 ,  urease  did not reduce virulence toward insects. The enhanced rhizoplane colonization of Δ Mep2  and Δ MepC  and insect derived nitrogen transfer to plant hosts suggests the role of  MepC  and  Mep2  in  Metarhizium -plant symbiosis.

plos one

Metarhizium robertsii ammonium permeases (MepC and Mep2) contribute to rhizoplane colonization and modulates the transfer of insect derived nitrogen to plants