Comparative genomics of Sympodiorosea identifies genome evolution mediated through selective pressure on the metabolic gene repertoire

Biological interactions involving host-associated fungi are manifestations of chemistry shaped over evolutionary time. For antagonistic fungi, specialization to host fungi is often facilitated through the acquisition of genes encoding novel secretions, including proteins and specialized metabolites that shape interactions with host defenses, as well the acquisition of nutrients. Alternatively, novel function may arise from diversifying selection eliciting evolutionary innovation in inert secretions. Through specialization, fungal metabolic capacity can serve as an imprint of the selective forces imposed on the antagonist and provide insights into its natural history and occupied niche. Here, we conduct a phylogenomic investigation of antagonistic ascomycetes in the genus Sympodiorosea, which are commonly found in basal fungus-growing ant gardens. Sympodiorosea and closely related genera of Escovopsis, Luteomyces, and Escovopsioides are canonically referenced as virulent mycoparasites, however, recent work has illuminated the possibility for diverse, non-virulent species to emerge within these attine-associated fungi. We explored genomic variation in Sympodiorosea to gain insight into the genomic potential for alternative lifestyles, focusing on diversity and evolution of metabolic genes. Our study revealed a constrained selective landscape across the Sympodiorosea genome. However, outcomes of in vitro interactions with host fungi are diverse and predictable based on the antagonists ant-species-of-origin, suggesting functional diversification. Phylogenomics reveals that gain/loss events of genes involved in secretions (secretome) are potential drivers of diversification. Additionally, we demonstrate that purifying selection acts more intensely on secretome-related genes relative to other genes. In contrast, few genes from the secretome experience diversifying selection, suggesting a mechanistic role for driving both functional differences between strains and host-specialization. Comparative genomics including other fungi within the family Hypocreales reveals that Sympodiorosea has experienced expansions and contractions in proteases genes that are discordant from expectations under a strictly mycoparasitic lifestyle, indicating either the potential for an alternative lifestyle within ant gardens, or that Sympodiorosea recently evolved from other niches. These results provide novel insight into genomic evolution of these fungi and inform future experimental studies of the ecology and chemistry of interactions within the complex fungus-growing ant symbiosis.