Towards identifying Brassica proteins involved in mediating resistance to Leptosphaeria maculans: a proteomics-based approach.
To better understand the pathogen-stress response of Brassica species against the ubiquitous hemi-biotroph fungus Leptosphaeria maculans, we conducted a comparative proteomic analysis between blackleg-susceptible Brassica napus and blackleg-resistant Brassica carinata following pathogen inoculation. We examined temporal changes (6, 12, 24, 48 and 72 h) in protein profiles of both species subjected to pathogen-challenge using two-dimensional gel electrophoresis. A total of 64 proteins were found to be significantly affected by the pathogen in the two species, out of which 51 protein spots were identified using tandem mass spectrometry. The proteins identified included antioxidant enzymes, photosynthetic and metabolic enzymes, and those involved in protein processing and signaling. Specifically, we observed that in the tolerant B. carinata, enzymes involved in the detoxification of free radicals increased in response to the pathogen whereas no such increase was observed in the susceptible B. napus. The expression of genes encoding four selected proteins was validated using quantitative real-time PCR and an additional one by Western blotting. Our findings are discussed with respect to tolerance or susceptibility of these species to the pathogen.
Sharma
Nidhi
N
Plant BioSystems & Agricultural Genomics and Proteomics Groups, Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada.
Hotte
Naomi
N
Rahman
Muhammad H
MH
Mohammadi
Mohsen
M
Deyholos
Michael K
MK
Kav
Nat N V
NN
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000472:pathogenicity*; D001937:Brassica / Q000737:chemistry*; D017343:Genes, Plant; D054884:Host-Pathogen Interactions / Q000502:physiology; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry
D010940:Plant Proteins