Studying Peroxisomes in Ustilago maydis: A Defined Workflow

Peroxisomes are highly conserved oxidative organelles essential for the breakdown of fatty acids and the detoxification of hydrogen peroxide (H₂O₂), a byproduct of this process. Beyond these central roles, peroxisomes contribute to diverse cellular functions, including amino acid biosynthesis and secondary metabolism. In recent years, the basidiomycete Ustilago maydis has emerged as a valuable model to uncover novel aspects of peroxisome biology. Here, we present a workflow to investigate the subcellular and suborganellar localization of peroxisomal proteins of interest and to assess their functional relevance. The outlined experimental strategies are broadly applicable to research questions beyond peroxisome biology and can be adapted to other systems.