Proteomic Techniques for PlantFungal Interactions

Proteomics is a key technique that is helping elucidate many complex biological processes. The analysis of plant–pathogen interactions using proteomics is complicated by the presence of the proteomes of two species, but is benefiting from the developing maturity and power of these techniques. More and more pathogen genomes are being sequenced, so fungal proteomics is reaching its full potential and remains the chosen technology to unravel the molecular pathways of pathogenicity and resistance. In this chapter, we suggest proteomic strategies that have proved successful on various plant-interacting fungal species. Several protein extraction methods are described. For adequate quantitative analyses of protein abundances, we recommend either separation using two-dimensional gel electrophoresis or labelling with isobaric tags followed by two-dimensional HPLC separation. Proteins of interest are then identified using mass spectrometry. Identified proteins can assist in refining genome annotations, otherwise known as proteogenomics.