SDS-PAGE of Ophioglossum vulgatum Proteins

Abstract

Background: Despite that the protein composition is one of the main characteristics of plant organism, there are not many studies dedicated to the fern proteins characterisation. Protein composition as a part of biochemical characterisation of Ophioglossum vulgatum (an evolutionary distinct fern species related to Psilotaceae) is of interest from the point of view of ecology, taxonomy and plant biochemistry. Aims: Study of the protein composition of roots and leaves of O. vulgatum during seasonal ontogenesis of its sporophytes. Methodology: SDS-PAGE with following densitometry was used for the estimation of protein composition of roots and leaves of O. vulgatum collected from young plants, mature plants, spore-bearing and not spore-bearing plants at the peak of their seasonal development. To determine the phosphorylation of studied proteins the electrophoresis using Phos-tag™ Acrylamide Gels were applied. Results: The simple and effective method of protein extraction from the leaves and roots of O. vulgatumwas developed and applied. Protein fractions with the molecular weights of 112±1.3, 57±1.0, 45±1.2, 35.5±0.6, 25.8±0.5, 18±1.5 kDa were shown as the major for both leaves and roots. Huge RuBisCO accumulation during the stages of plant initial growth and spore-bearing was shown (up to 55±10% in leaves and 89±5% in roots). All large chains of detected RuBisCO were phosphorylated which suggests that the active form of O. vulgatum RuBisCO is phosphoprotein. Conclusion: The dynamic change of protein content was shown for different ontogenesis stages of O. vulgatum. The expression of phosphorylated RuBisCO was distinctly increased during the beginning of leaf development and spore-bearing.