Manipulating multi-level selection in a fungal entomopathogen reveals social conflicts and a method for improving biocontrol traits
Authors: Erdos Zoltan , Studholme David J. , Sharma Manmohan D , Chandler David , Bass Chris , Raymond Ben 
Published: November 2023

Changes in parasite virulence are commonly expected to lead to trade-offs in other life history traits that can affect fitness. Understanding these trade-offs is particularly important if we want to manipulate the virulence of microbial biological control agents. Theoretically, selection across different spatial scales, i.e. between- and within-hosts, shapes these trade- offs. However, trade-offs are also dependent on parasite biology. Despite their applied importance the evolution of virulence in fungal parasites is poorly understood: virulence can be unstable in culture and commonly fails to increase in simple passage experiments. We hypothesized that manipulating selection intensity at different scales would reveal virulence trade-offs in a fungal pathogen of aphids,Akanthomyces muscarius. Starting with a genetically diverse stock we selected for infectivity, parasite yield or speed of kill by manipulating competition within and between hosts and between groups of hosts over 7 rounds of infection. We characterized ancestral and evolved lineages by whole genome sequencing and by measuring virulence, growth rate, sporulation and fitness. While several lineages showed increases in virulence, we saw none of the trade-offs commonly found in obligately-killing parasites. Phenotypically similar lineages within treatments often shared multiple single-nucleotide variants, indicating strong convergent evolution. The most dramatic phenotypic changes were in timing of sporulation and spore productionin vitro.We found that early sporulation led to reduced competitive fitness but could increase yield of spores on media, a trade-off characteristic of social conflict. Notably, the selection regime with strongest between-group competition and lowest genetic diversity produced the most consistent shift to early sporulation, as predicted by social evolution theory. Mutli-level selection therefore revealed social interactions novel to fungi and showed that these biocontrol agents have the genomic flexibility to improve multiple traits -virulence and spore production - that are often in conflict in other parasites.

Author summary

Understanding the ecological forces that shape virulence is a key challenge in evolutionary biology. Here we investigated how competition at different levels of selection (within-hosts, between-hosts, between populations) could alter investment in virulence in a fungal entomopathogen. We predicted that cooperative investment in virulence would increase at higher scales of competition and aimed to further our understanding of potential trade-offs shaping life-history of a fungal insect pathogens. We found moderate increases in virulence in different selection regimes but importantly, none of the commonly expected trade-offs, such as that between spore production and virulence or a relationship between virulence and growth rate, which is consistent with cooperation. However, we found convergent genetic changes and significant differences in timing and production of spores, dependent how we manipulated scales of selection. Our data suggests that this is driven by social conflict regarding the timing of sporulation. This carries fundamental importance for understanding how varying selection pressure at different scales shape pathogen life history. In addition, these results also have applied importance for understanding how to improve and select for beneficial traits in biocontrol agents.