Proteomic Analysis of Somatic Embryogenesis in Cyclamen persicum Mill

Abstract

Cyclamen persicum Mill. is a widely grown ornamental species that is clonally propagated by somatic embryogenesis. To better understand the biology of somatic embryo development in C. persicum, detailed proteomic (two-dimensional gel electrophoresis) and mass spectrometric analyses of somatic embryos at globular, torpedo, and germinating stages of development, along with nonembryogenic callus and zygotic embryos, were conducted. Of~460 proteins resolved in two-dimensional gels, 35 proteins were differentially expressed and could be reproducibly displayed across an isoelectric focusing range of 5 to 8. Among those proteins, five were constitutively expressed, 13 were upregulated, nine were downregulated, and eight were deemed as novel proteins during the torpedo stage. A total of 35 protein spots were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and only four proteins were identified and these were available in public protein databases. The remaining protein spots were subsequently analyzed by MALDI-TOF-TOF-MS, and six proteins were then identified. These findings suggested that specific proteins are involved in the regulation of somatic embryogenesis.

Proteomic Analysis of PEG-Fractionated UV-C Stress-Response Proteins in Globe Artichoke

“…The experimental samples were isoelectrofocused together in the same run to reduce technical variability. The IPG strips were then incubated in 4 mL of equilibration buffer [6 M urea, 30% (w/v) glycerol, 2% (w/v) sodium dodecyl sulphate (SDS), 0.002% (w/v) bromophenol blue, 50 mM Tris-HCl, pH 8.8] containing 1% (w/v) DTT for 15 min and subsequently in 4 mL of the same buffer containing 2.5% (w/v) iodoacetamide (IAA) for 15 min and subjected to SDS polyacrylamide gel electrophoresis in a 12.5% polyacrylamide gel adopting the running conditions reported in Bian et al (2010). The 2-DE gels were stained with Sypro Ruby (BioRad's) following manufacturer's instructions.…”

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

“…We did not observe the unique differences in the number of produced protein spots between stage III, VI and VII with the number about 545. In the case of Cyclamen persicum, studies revealed that the differing number of spots between particular stages is not large, and globular somatic embryo stage was characterized by 451 protein spots while the torpedo stage was characterized by 460 protein spots [11].…”

“…For both extreme stages, some proteins had acid character just about pH 4.0 or a base character just between pH values ranging from 9 to 10. Globular and torpedo somatic embryos of C. persicum presented IEF of proteins ranging pH 5-8, only [11].…”

“…Somatic embryogenesis is a complex developmental process that offers multidisciplinary studies of embryogenesis and great potential in plant propagation. Since two-dimensional electrophoresis (2-DE) was developed the application of the method has been increasingly used as a tool for studies of somatic embryogenesis [10][11][12]. 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].…”

The Current Status of Proteomic Studies in Somatic Embryogenesis

“…Furthermore, proteomic studies of SE in several species have created protein profiles of cultures in different conditions during the induction of embryogenic potential (Varhanikova et al 2014;Correia et al 2012;Marsoni et al 2008;Sharifi et al 2012;Almeida et al 2012;Guzman-Garcia et al 2013). These studies have been followed up with the focused identification of key proteins associated with each stage of the transdifferentiation of somatic embryos (Rode et al 2012;Bian et al 2010;Vale Ede et al 2014). Moreover, the morphological and physiological similarities between zygotic and somatic embryogenesis have suggested similar biochemical status for cells undergoing both of these processes, and protein profiles for zygotic and somatic embryogenesis have been compared in a few plants species (Sghaier-Hammami et al 2009;Winkelmann et al 2006;Noah et al 2013).…”