Proteome-level investigation of Brassica carinata-derived resistance to Leptosphaeria maculans

Plants resistant to the fungal pathogen Leptosphaeria maculans were generated by an interspecific cross between the highly susceptible Brassica napus (canola) and the highly resistant Brassica carinata. Changes in the leaf protein profiles of these lines were investigated in order to understand the biochemical basis for the observed resistance. Two-dimensional electrophoresis followed by tandem mass spectrometry led to the identification of proteins unique to the susceptible (5 proteins) and resistant genotypes (7 proteins) as well those that were differentially expressed in the resistant genotype 48 h after challenge with the pathogen (28 proteins). Proteins identified as being unique in the resistant plant material included superoxide dismutase, nitrate reductase, and carbonic anhydrase. Photosynthetic enzymes (fructose bisphosphate aldolase, triose phosphate isomerase, sedoheptulose bisphosphatase), dehydroascorbate reductase, peroxiredoxin, malate dehydrogenase, glutamine synthetase, N-glyceraldehyde-2-phosphotransferase, and peptidyl-prolyl cis-trans isomerase were observed to be elevated in the resistant genotype upon pathogen challenge. Increased levels of the antioxidant enzyme superoxide dismutase were further validated and supported by spectrophotometric and in-gel activity assays. Other proteins identified in this study such as nitrate reductase and peptidylprolyl isomerase have not been previously described in this plant-pathogen system, and their potential involvement in an incompatible interaction is discussed.