Primary results of 2-dimensional electrophoresis for protein studies of Gentiana kurroo Royle somatic embryos derived from long-term embryogenic cell suspensions

Abstract

Somatic embryos were selected from hypocotyl derived suspension culture (Fig. 1a) described previously [7] and used for protein isolation and detection (Fig. 2). Different Abstract Two-dimensional gel electrophoresis was used to compare protein expression profiles between various stages of Gentiana kurroo Royle somatic embryos. Seven distinct stages (I-VII) were pointed out and measured from long-term embryogenic cell suspension. Isoelectric focusing was done in the pH intervals 3-10, and the second dimension was carried out with 13% SDS-polyacrylamide gel electrophoresis. Dependent on the stage from about 400 (stage IV) to more than 700 (stage II) protein spots were totally detected. The molecular weight of abundant proteins range from 12 to 70 kDa, however, majority of proteins were located between 20-49 kDa spots on the gels. The highest difference in the number of spots appeared in the case of globular embryo (stage I) and elongated cotyledonary stage (stage VII) with differences being about 130 spots. The relevance of embryogenic cell suspension choice for proteomic analysis as well as expediency of the increasing number of particular embryo stages is discussed.

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

“…Attention was paid to the recognition of the number of protein spots between particular stages. The differences ranged from the lowest number of the spots presented by Stage IV to the highest number of spots presented in Stage II, with over 300 different protein spots (Niedziela and Rybczyński, 2014).…”

Electroporation and morphogenic potential of Gentiana kurroo (Royle) embryogenic cell suspension protoplasts

“…A very high yield of regenerants via somatic embryogenesis from proto- plast bead culture has confirmed the usefulness of this type of protoplast culture for gentian cell manipulation (Fiuk and Rybczyński, 2007). The cell suspension cultures also produce embryogenic aggregates and somatic embryos which could be used for proteomic studies (Fiuk and Rybczyński, 2007;Niedziela and Rybczyński, 2014). Somatic embryogenesis is a complex process that allows the performance of multidisciplinary studies into the mechanism of embryo induction and its development.…”