Isolation of Mitochondria from Fusarium Species

Isolation of purified mitochondria is an essential technique for the analysis of metabolic and cellular functions associated with this vital organelle. Filamentous fungi, such as Neurospora crassa, have been proven to be highly amenable to the analysis of mitochondria, in part due to their rapid growth rate and relative ease of isolation. Here we describe a step-by-step procedure for the isolation of mitochondria from Fusarium species via differential centrifugation and density step-gradient centrifugation, and include methods to overcome potential complications. Mitochondria purified by flotation gradient procedures remain active for functional assays and can be further fractionated for isolation of nucleic acids or ribonucleoprotein particles that retain enzymatic activity.