Abstract
Two-dimensional gel electrophoresis was used to compare protein expression profiles between various stages of <em>Gentiana kurroo</em> 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.
Highlights
Gentianaceae family consists about 1700 species, but only 28 are the object of plant tissue culture and biotechnology
The very first paper showed possibilities to establish plant multiplication of Gentiana kurroo by culturing the shoot tips and nodal segments on Murashige and Skoog (MS) medium supplemented with BAP and NAA [3]
Two-dimensional gel electrophoresis has been successfully engaging in proteome studies of encapsulated embryogenic cell suspension during its adaptation to osmotic stress in cryopreservation protocol of Gentiana cruciata [16]
Summary
Gentianaceae family consists about 1700 species, but only 28 are the object of plant tissue culture and biotechnology. The first papers summarizing achievements of gentian plant tissue cultures were published in 1988 and 1991 [1,2]. The very first paper showed possibilities to establish plant multiplication of Gentiana kurroo by culturing the shoot tips and nodal segments on Murashige and Skoog (MS) medium supplemented with BAP (benzylaminopurine) and NAA (naphtaleneacetic acid) [3]. The significant progress in vegetative plant cell manipulation was recognized when the somatic embryogenesis of primary explants for a few gentian species was described [5]. G. kurroo appeared very embrygenic with tremendous morphogenic potential since seedling and leaf explants [6] were used for culture initiation. It should be stressed that proteomic study in gentians has not been published until now, with the exception of one describing isoenzymes and protein patterns in leaves of in vitro micropropagated plantlets of G. lutea in the presence of various combination of plant growth regulators [13]
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