Introduction: Aleurone layer (AL), is a living cell layer among the peripheral layers of the grain structure obtained after milling wheat, is rich in, vitamins, minerals, and antioxidants potentially nutritional value of the flour. Objectives: to isolate AL in mature grain of the three major species Common wheat (CW), Durum wheat (DW), and Eikinon wheat (EW) that were grown at two different years as well as to analyze and compare their proteomes revealed through two-dimensional electrophoresis (2DE) and image analysis. Methods: The AL was hand dissected and unicellular purity verified using scanning electron microscopy. AL proteins were separated using IEF pH3-10 X SDS-PAGE then Coomassie stained, the gels were scanned and the images were compared using Samespot (Nonlinear Dynamics) and were proteins identified using mass spectrometry and database interrogation. Results: For CW and DW samples, no significant quantitative or qualitative differences was observed between the AL proteome in the two years. However, a few quantitative differences were revealed between EW for AL in 2006 and 2007. The proteins identified were classified in the carbohydrate pathway and stress defense response. Conclusion: This remarkable stability over environmental growing conditions strengthens need to pay a greater attention to this unicellular living cell of the wheat grain.
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