# An Integrative Multi-Source Evidence Framework for Prioritizing Virulence-Associated Pathways in Metarhizium brunneum

## TL;DR

This paper introduces a new framework to identify key virulence pathways in the fungus Metarhizium brunneum, which could improve its use as a biocontrol agent against pests.

## Contribution

The study presents an integrative multi-source evidence framework for prioritizing virulence pathways in entomopathogenic fungi.

## Key findings

- Five pathways, including MAPK signaling and apoptosis, were identified as key virulence-related candidates.
- The framework combines protein pathway enrichment, GO analysis, PHI-base mapping, and literature evidence.
- The approach is scalable and can be applied to other fungi to optimize biocontrol strategies.

## Abstract

Background: The entomopathogenic fungus Metarhizium brunneum (M. brunneum) is an effective biocontrol agent against various vector arthropods such as ticks, mosquitoes, and flies. However, its virulence mechanisms remain poorly understood, which hinders its broader application. This study aims to establish an integrative framework for prioritizing virulence-related pathways in M. brunneum to aid in the development of more effective biocontrol strategies. Methods: A multidimensional virulence pathway scoring framework was developed using publicly available protein annotation data of M. brunneum. This approach integrates protein pathway enrichment, Gene Ontology (GO) functional analysis, PHI-base virulence factor mapping, and literature-derived evidence. A total of 20 pathways were evaluated, and a scoring system was applied based on protein coverage, Gene Ontology Biological Process (GO-BP) support, PHI-base hits, and literature support. Results: Among the 20 pathways evaluated, five pathways, including MAPK signaling, apoptosis, endocytosis, carbon metabolism, and biosynthesis of secondary metabolites received the highest priority scores. These pathways were identified as key virulence-related candidates, supported by both functional annotation and existing biological evidence. Conclusions: The proposed framework provides a reliable and scalable strategy for prioritizing virulence pathways in entomopathogenic fungi. It offers a solid foundation for subsequent transcriptomic validation, target screening, and functional characterization. This framework can also be applied to other fungi, contributing to the development of optimized biocontrol formulations.