A biosynthetic survey of biocontrol fungi

<title>Abstract</title> Pests are responsible for the loss of up to 40% of global crops. Widespread resistance to pesticides has emerged due to the overuse of a limited set of compounds and targets. This overreliance has driven the rapid adaptation of pest populations, making pesticides increasingly ineffective while posing significant risks to public health and the environment. Many fungal species form symbiotic relationships with plants while antagonizing insects, nematodes, and bacteria, making them valuable sources of sustainable biocontrol agents and biopesticides for agriculture. However, despite their potential, few of these species are used in biocontrol. Furthermore, little is known about their biosynthetic capabilities. Here we used phylogenomics, metabolomics, and heterologous expression to catalog and characterize the biosynthetic repertoire of biocontrol species within the Hypocreales order. Our workflow, applied to 82 plant- and insect-associated Hypocreales species, led to the annotation of 5221 biosynthetic gene clusters and putative products hitherto for 1034 of them. By linking biosynthetic gene clusters to molecules, we investigated the biosynthesis of several natural products. Additionally, by combining our metabologenomics workflow with synthetic biology we characterized 4 NRPS-like synthetases involved in the biosynthesis of hitherto unknow products. We believe this biosynthetic survey will provide a valuable foundation for developing sustainable pest control strategies in agriculture.