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Lysin motif (LysM) modules are approximately 50 amino acids long and bind to peptidoglycan, chitin and its derivatives. Certain LysM proteins in plant pathogenic and entomopathogenic fungi are shown to scavenge chitin oligosaccharides and thereby dampen host defense reactions. Other LysM proteins can protect the fungal cell wall against hydrolytic enzymes. In this study, we investigated the biological function of LysM proteins in the mycoparasitic fungus  Clonostachys rosea . The  C .  rosea  genome contained three genes coding for LysM-containing proteins and gene expression analysis revealed that  lysm1  and  lysm2  were induced during mycoparasitic interaction with  Fusarium graminearum  and during colonization of wheat roots.  Lysm1  was suppressed in germinating conidia, while  lysm2  was induced during growth in chitin or peptidoglycan-containing medium. Deletion of  lysm1  and  lysm2  resulted in mutants with increased levels of conidiation and conidial germination, but reduced ability to control plant diseases caused by  F .  graminearum  and  Botrytis cinerea . The Δ lysm2  strain showed a distinct, accelerated mycelial disintegration phenotype accompanied by reduced biomass production and hyphal protection against hydrolytic enzymes including chitinases, suggesting a role of LYSM2 in hyphal protection against chitinases. The Δ lysm2  and Δ lysm1 Δ lysm2  strains displayed reduced ability to colonize wheat roots, while only Δ lysm1 Δ lysm2  failed to suppress expression of the wheat defense response genes  PR1  and  PR4 . Based on our data, we propose a role of LYSM1 as a regulator of fungal development and of LYSM2 in cell wall protection against endogenous hydrolytic enzymes, while both are required to suppress plant defense responses. Our findings expand the understanding of the role of LysM proteins in fungal-fungal interactions and biocontrol.

LysM Proteins Regulate Fungal Development and Contribute to Hyphal Protection and Biocontrol Traits in Clonostachys rosea