Proteomics research for plant self defense proteins

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To investigate the proteome analysis and inherent self defense proteins, the inducible self defense proteins were subjected to proteomic analysis. The method for 2D electrophoresis of plant tissue extract, in-gel digestion, and mass spectrometry were established and applied to the detection of chitinase induced by stress followed by identification of the proteins using mass spectrometry. The zymographic procedure was also developed for plant chitinase and analyzed by mass spectrometry using optimized method.The hydroponic cultured ruzigrass under Al stress was analyzed its inducible proteins on 2D electrophoresis. The detected protein spot was analyzed on mass spectrometry after in-gel digestion of the protein using established protocol. The protein was identified as class 1 chitinase family, however, carried higher molecular weight than normal class 1 chitinase. This suggests that the detected induced chitinase was newly found class 1 type chitinase that carried multiple chitin binding domains. The extract of ruzigrass roots was then applied on to an affinity column using Chitipearl and separated adsorbed chitinase fractions.To elucidate the identification of inducible chitinase during infection or wounding, yam aerial tuber infected by fungi and wounded by insects were subjected zymographic analysis of substrate gel SDS electrophoresis. The results indicate that the chitinase apparently induced by fungal infection and wounding.