Abstract:

【Objective】 A trichoroacetic acid/acetone method was established for extracting the full leaf-proteins of rapeseed which provided the reference for the rapeseed proteomics research in the future. The difference in protein expression was identified after the resistance and susceptibility to Sclerotinia sclerotiorum rapeseed were induced by Sclerotinia sclerotiorum. It offered the clues to searching for the resistance to Sclerotinia sclerotiorum genes.【Method】 Near-isogenic lines 98C40 (the antigen derived from Xiangyou 15) is resistant to Sclerotinia sclerotiorum and the parent 98C40 is susceptible to Sclerotinia sclerotiorum which were inoculated with Sclerotinia sclerotiorum during their seedling stage. Seventy-two hours after inoculation, full leaf-proteins of them were extracted respectively. Two-dimensional electrophoresis (2-DE), PD-quest software and mass technology were used in differential proteomics analysis. 【Result】 Significant difference of 4 protein spots were observed between them. Protein spot a is mitochondrial F1 ATP synthase beta subunit, spot c is phosphoglycerate kinase and spot d is thiazole biosynthetic enzymes (THI1), which express only in 98C40 (the antigen derived from Xiangyou 15) not expressed in parent 98C40. Protein spot b is ribulose-1,5-bisphosphate carboxylase/oxygenase expressed highly in 98C40 (the antigen derived from Xiangyou 15) and lowly in parent 98C40 based on 5 times difference of expression. 【Conclusion】 The four proteins are all closely related to the rapeseed metabolism. They may play a role in resistance to Sclerotinia sclerotiorum.

Key words: rapeseed, Sclerotinia sclerotiorum, proteomics, near-isogenic lines, two-dimensional electrophoresis, mass spectrometry