Analysis of Secreted Proteins from Macrophomina phaseolina Isolated from Infected Glycine max Tissues

Necrotrophic fungi such as Macrophomina phaseolina (MP) pose a growing threat to global food security, especially as climate change expands their ecological range. MP is a highly adaptable phytopathogen that secretes a diverse array of metabolites and proteins-collectively known as the secretome-which play critical roles in host colonization and disease progression. In this chapter, we describe a method to evaluate the secretome of MP stimulated with ground Glycine max roots and seedlings, to identify proteins potentially implicated in the infection process. Current methods for analyzing proteins secreted by Mp do not incorporate supplementation with host plant tissue. However, including plant-derived substrates in secretion assays could stimulate the production of phytotoxic proteins that are otherwise not detected under standard conditions. These secreted effectors may play key roles in pathogenesis, and their function can be further validated through knockout mutagenesis or heterologous expression studies.