Enhanced Protein Extraction from Tobacco Roots for Proteomic Analysis

Abstract

Plant roots contain low protein concentrations and many interferences for protein extraction and two-dimensional electrophoresis analysis. Therefore, the extraction of high-quality protein from tobacco roots for proteomic analysis is a challenge. Three protein extraction methods (the trichloroacetic acid-acetone, phenol extraction, and trichloroacetic acid-acetone-phenol methods) for tobacco root proteins were compared using protein yields, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and two-dimensional electrophoresis. The trichloroacetic acid-acetone-phenol method provided a higher spot resolution (505 ± 18 spots), the least streaking, and larger protein yields (2200 ± 20 µg/g fresh weight) on two-dimensional electrophoresis gels for tobacco roots, and hence is the most suitable method for the characterization of tobacco roots.