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A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample
Published on: June 1, 2017
Integrated proteomics and metabolomics analysis of chickpea seeds under aging condition
Rinku Malviya1, Shuvobrata Majumder2, Punam Sharma1
1Department of Biochemistry, Central University of Rajasthan, Ajmer 305817, India.
Abstract:
Seed aging in chickpeas during storage highlights significant economic and agricultural concerns. In this study, chickpea seeds were aged at 45 °C and 85 % humidity for 4 days, which affected both seeds' viability and vigor. Comparative biochemical analyses showed a higher production of ROS and membrane damage in the aged seeds. Comparative proteomics and metabolomics analyses of seeds were conducted using Easy-nLC LC-MS/MS and GC-MS techniques. This examination identified 2605 proteins, 17654 peptides, and 68 metabolites between the control and aged seeds. A total of 118 statistically significant proteins were identified, with 98 proteins exhibiting differential abundance in response to the aging treatment. The in-silico functional and subcellular analysis revealed that these proteins and metabolites are related to germination, desiccation tolerance, phytohormones, fatty acid, amino acid metabolism, carbon assimilation pathways, and exhibited several locations inside the cell. The relative expression of 8 selected genes was verified using qRT-PCR to check the pattern with proteomics results. Temperature-induced lipocalin (CaTIL) is a protective protein that stabilizes the membrane and provides tolerance to aging, and was selected for molecular characterization. The subcellular localization of CaTIL was found to be in the plasma membrane of the onion cells.

