Methods to Detect Apoptotic-Like Cell Death in Filamentous Fungi

Fungi are capable of undergoing apoptotic-like cell death, and display many of the characteristic features of apoptosis observed in multicellular organisms. These features include nuclear condensation, DNA fragmentation, translocation of phosphatidylserine from the cytoplasmic to the extracellular side of the plasma membrane, and increased levels of reactive oxygen species (ROS). Several assays can be used to detect apoptotic cells, and here we describe adaptations of assays such as TUNEL, Annexin V, and Evan's Blue for the investigation of apoptotic-like cell death in fungal hyphae. We also present approaches for monitoring nuclear condensation and production of ROS.