Analysis Of Protein Expression Profiling In Rice Disease Resistance Gene-Mediated Resistance

Proteomics covers the systematic analysis of proteins expressed by a genome, from the identification of their primary amino acid sequences to the determination of modification and association with other proteins or molecules of different types. Various analyse techniques of protein expression have been used in the field of plant accompany with the promotion of proteomics. Rice has immense socio-economic impact on human existence. However, rice disease dramatically limits the development of rice industry. Up to now, the exact genome sequence of rice has been released. Many progresses have been made with the proteomic techniques such as two-dimensional electrophoresis and mass spectrum in the field of rice metabolisms and regulations.In this study, a complete analytical system, including rice leaf protein extraction, protein quantification, two dimensional electrophoresis, silver staining, dots pick, electrospray ionization mass spectrometry (ESI-MS) detection and data analysis, was established. In this system, a novel mass spectrometry-compatible silver staining method for two dimensional electrophoresis gel with low-background and high resolution was developed. We analyzed the protein expression profiling during rice-Xanthomonas oryzae pv oryzae (Xoo) interaction by using a two-dimensional electrophoresis-mass spectrometry strategy. Proteins were extracted from leaves at 1, 4, 8, 48, and 72 hours after Xoo inoculation. Seventy-two differentially expressed protein spots were identified by comparing the expression profilings of proteins from Xoo- and mock-inoculated plants revealed by two-dimensional electrophoresis. Further analysis using electrospray ionization-time of flight mass spectrometry revealed the property of 11 of the 72 differentially expressed proteins. They are: RNA binding protein, succinyl-CoA ligase alpha subunit, ribulose 1,5-bisphosphate carboxylase/oxygenase, no apical meristem protein, leaf senescence protein-like, carbonic anhydrase, transducin family protein/ WD-40 repeat family protein, acetolactate synthase, glyceraldehyde 3-phosphate dehydrogenase B subunit and cytosolic glyceraldehyde 3-phosphate dehydrogenase. These results provide basis for further study of the molecular mechanism of rice disease resistance.