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A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
Published on: January 25, 2018
Organ-Specific Proteomic Analysis of Early Vegetative Stage Soybean
Mudassar Nawaz Khan1, Setsuko Komatsu1
1Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572. Japan.
Background:
Soybean is a nutritionally important source of proteins and oils. Organspecific analyses of the plants provided valuable information about physiological changes occurring in particular organ. However, studies on the vegetative stage soybean during the growth and development are lacking. Analyzing the growth stages of soybean at proteomic level is particularly important for understanding the growth dynamics.
Objectives:
The main objective of this study was to investigate the organ-specific growth pattern of proteins during early growth stage.
Methods:
Soybean (Glycine max L. cv Enrei) seeds were sown in each seedling case for 10 days and roots, hypocotyls, and leaves were collected. Proteins were extracted and analyzed by nano-liquid chromatography mass spectrometry. Mole percent abundance was calculated using emPAI values. To determine the functional role of the proteins identified in the MS analysis, functional categorization was performed using MapMan bin codes. Visualization of protein abundance ratio was performed using MapMan software. Enzyme activity and quantitative reverse transcription-polymerase chain reaction analyses were performed.
Results:
A total of 357, 360, and 392 proteins were identified in root, hypocotyl, and leaf of vegetative stage soybean, respectively. Proteins related to stress, cell organization, transport, signaling, and mitochondrial electron transport chain decreased in root, hypocotyl, and leaf. Proteins related to protein metabolism, glycolysis, and cell wall were comparable in root and hypocotyl; however, in leaf, glycolysis and cell wall related proteins were decreased. Aldehyde dehydrogenase was decreased in abundance and activity in hypocotyl and leaf as compared to root. Major latex proteins 43 and 423 changed in abundance in an organ-specific manner. The mRNA expression level of major latex proteins exhibited a differential expression in the hypocotyl of soybean during flooding stress and recovery.
Conclusion:
Our results suggest that aldehyde dehydrogenase and major latex proteins play key roles in growth of soybean in an organ-specific way.
