Early diverging insect pathogenic fungi of the order Entomophthorales possess diverse and unique subtilisin-like serine proteases

Abstract

Insect-pathogenic fungi use subtilisin-like serine proteases (SLSPs) to degrade chitin-associated proteins in the insect procuticle. Most insect-pathogenic fungi in the order Hypocreales (Ascomycota) are generalist species with a broad host-range, and most species possess a high number of SLSPs. The other major clade of insect-pathogenic fungi is part of the subphylum Entomophthoromycotina (Zoopagomycota, formerly Zygomycota) which consists of high host-specificity insect-pathogenic fungi that naturally only infect a single or very few host species. The extent to which insect-pathogenic fungi in the order Entomophthorales rely on SLSPs is unknown. Here we take advantage of recently available transcriptomic and genomic datasets from four genera within Entomophthoromycotina: the saprobic or opportunistic pathogens Basidiobolus meristosporus, Conidiobolus coronatus, C. thromboides, C. incongruus, and the host-specific insect pathogens Entomphthora muscae and Pandora formicae, specific pathogens of house flies (Muscae domestica) and wood ants (Formica polyctena), respectively. We use phylogenetics and protein domain analysis to show that the obligate biotrophic fungi E. muscae, P. formicae and the saprobic human pathogen C. incongruus all contain \"classical\" fungal SLSPs and a unique group of SLSPs that loosely resembles bacillopeptidase F-like SLSPs. This novel group of SLSPs is found in the genomes of obligate insect pathogens and a generalist saprobic opportunistic pathogen why they are unlikely to be responsible for the host specificity of Entomophthorales. However, this class represent a unique group of SLSPs so far only observed among Bacteria, Oomycetes and early diverging fungi such as Cryptomycota, Microsporidia, and Entomophthoromycotina and missing in the sister fungal lineages of Kickxellomycotina or the fungal phyla Mucoromyocta, Ascomycota and Basidiomycota fungi suggesting interesting gene loss patterns.