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      • Shotgun proteomics approach to identify candidate proteins related with drought stress in Brassica rapa ssp. pekinensis (inbred line ‘Chiifu’)

        Hijin Kim,Jaebok Cho,Wondo Lee,Joohyun Lee 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07

        As the drought is getting worse, Lot of studies related to drought stress in plant have been conducted. Recently whole genome sequencing of Brassica rapa ssp which is important vegetable crop to East Asians has been completed to enable Omic research. It is known that the drought damages occur in the early stage of plant development. Here, we performed shotgun proteomics analysis of B. rapa to observe the morphological characters, monitor the expression patterns of the identified proteins during drought stress, and detect the proteins related to drought stress. The three week old B. rapa grown in density of single plant in a single pot were used. Drought stress were treated as that a single plant in soil was removed from the pot and the plant with soil was exposed to air and light without watering. Leaves were immediately harvested before drought treatment, 24hr after drought treatment, and 48hr after drought treatment. The protein expression patterns were monitored by a quantitative shotgun proteomics analysis. Extracted proteins were separated in 1D-SDS-PAGE then the gel sliced into seven pieces. Chopped gels were ingel-digested. Peptides were assigned to mass spectrometry (Q-Exactive). The ms/ms spectra were analyzed through Proteome Discoverer. By combining all of the identified proteins in the seven sliced gel samples, total B. rapa proteome reference map was completed. Protein expression patterns were investigated by comparing the quantity of protein. With shotgun proteomic approach, we evaluated the changes in the quantity and finally discovered the candidate proteins related with drought stress.

      • A quantitative shotgun proteomic study of rice grain development

        Wondo Lee,Jaebok Cho,Hijin Kim,Joohyun Lee 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

        Although a great deal of rice proteomic research has been conducted, there are relatively few studies specifically addressing the rice grain proteome. The existing rice grain proteomic research has focused on the identification of deferentially expressed proteins. Here, we performed comparative shotgun proteomic analysis of rice grain development to construct an in-depth proteome reference map, to reveal the expression patterns of the identified proteins, and to detect proteins that are expressed deferentially during grain development. A Korean rice variety, Ilpumbyeo was used. Proteins were extracted from rice grains 10, 20, and 30 days after flowering, as well as from mature grains. The protein expression patterns were revealed by a quantitative shotgun proteoemic analysis. By merging all of the identified proteins in this study, we identified 4,172 non-redundant. A Genome Ontology category enrichment analysis for the 4,172 proteins revealed that 52 categories were enriched, including the carbohydrate metabolic process, transport, localization, lipid metabolic process, and secondary metabolic process. The relative abundances of the 1,784 reproducibly identified proteins were compared to detect 484 differentially expressed proteins during rice grain development. Clustering analysis and Genome Ontology category enrichment analysis revealed that proteins involved in the metabolic process were enriched through all stages of development, suggesting that proteome changes occurred even in the desiccation phase. Interestingly, enrichments of proteins involved in protein folding were detected in the desiccation phase and in fully mature grain.

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