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Comparative Proteomics of Seed Maturation in Oil Seeds
Author(s) -
Thelen Jay,
Stevenson Severin,
Wilson Rashaun,
Salie Matthew,
Rao Shyama
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.220.2
Subject(s) - endosperm , rapeseed , proteome , proteomics , biology , storage protein , embryo , arabidopsis , microbiology and biotechnology , phosphorylation , computational biology , protein phosphorylation , biochemistry , botany , protein kinase a , mutant , gene
Oilseeds are renewable sources of protein and oil, which are produced primarily during the maturation phase of embryo development. Systems biological approaches to study seed development and metabolism have helped to resolve pathways for conversion of photoassimilate into storage reserves. Beginning in 2004, my lab employed “top‐down” proteomic approaches, specifically high‐resolution two dimensional (2D) gels, to resolve and monitor steady state protein levels and protein phosphorylation at various stages of seed development in soybean, rapeseed, castor, and Arabidopsis. This approach allowed for the profiling of roughly the 1,000 most abundant proteins, which included most of the enzymes in involved in primary/intermediary metabolic pathways required for assimilation of leaf sucrose into seed storage reserves. We propose oilseed‐specific differences in intermediary metabolism based upon these steady state expression models and show examples of proteins that undergo protein phosphorylation, which accounts for some of the 2D gel spot diversity. To further interrogate the developing rapeseed proteome we recently employed a “bottom‐up” strategy called GeLC‐mass spectrometry (MS) whereby proteins are resolved by preparative SDS‐PAGE and subsequently digested into peptides for tandem MS analysis and spectral counting. Using this approach to perform a spatio‐temporal analysis of rapeseed we identified 8,766 non‐redundant proteins from dissected seed coat, endosperm, embryo, and funiculi (maternal) tissues at multiple stages of development. The results of this study will be presented.