Proteomic analysis of stipe explants reveals differentially expressed proteins involved in early direct somatic embryogenesis of the tree fern Cyathea delgadii Sternb.
Using cyto-morphological analysis of somatic embryogenesis (SE) in the tree fern Cyathea delgadii as a guide, we performed a comparative proteomic analysis in stipe explants undergoing direct SE. Plant material was cultured on hormone-free medium supplemented with 2% sucrose. Phenol extracted proteins were separated using two-dimensional gel electrophoresis (2-DE) and mass spectrometry was performed for protein identification. A total number of 114 differentially regulated proteins was identified during early SE, i.e. when the first cell divisions started and several-cell pro-embryos were formed. Proteins were assigned to seven functional categories: carbohydrate metabolism, protein metabolism, cell organization, defense and stress responses, amino acid metabolism, purine metabolism, and fatty acid metabolism. Carbohydrate and protein metabolism were found to be the most sensitive SE functions with the greatest number of alterations in the intensity of spots in gel. Differences, especially in non-enzymatic and structural protein abundance, are indicative for cell organization, including cytoskeleton rearrangement and changes in cell wall components. The highest induced changes concern those enzymes related to fatty acid metabolism. Global analysis of the proteome reveals several proteins that can represent markers for the first 16days of SE induction and expression in fern. The findings of this research improve the understanding of molecular processes involved in direct SE in C. delgadii.
Polish Academy of Sciences Botanical Garden, Center for Biological Diversity Conservation in Powsin, Prawdziwka 2, 02-973 Warsaw, Poland.
Lucyna
L
Domżalska
Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Sylwia
S
Kędracka-Krok
Department of Structural Biology, Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.
Urszula
U
Jankowska
Polish Academy of Sciences Botanical Garden, Center for Biological Diversity Conservation in Powsin, Prawdziwka 2, 02-973 Warsaw, Poland.
Małgorzata
M
Grzyb
Department of Botany, Warsaw University of Life Sciences (SGGW), Warsaw, Poland.
Mirosław
M
Sobczak
Polish Academy of Sciences Botanical Garden, Center for Biological Diversity Conservation in Powsin, Prawdziwka 2, 02-973 Warsaw, Poland.
Jan J
JJ
Rybczyński
Polish Academy of Sciences Botanical Garden, Center for Biological Diversity Conservation in Powsin, Prawdziwka 2, 02-973 Warsaw, Poland. Electronic address: amikula@obpan.pl.
Anna
A
Mikuła
D004591:Electrophoresis, Polyacrylamide Gel; D029624:Ferns; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D013058:Mass Spectrometry; D046529:Microscopy, Electron, Transmission; D010940:Plant Proteins / Q000378:metabolism* / Q000502:physiology; D058439:Plant Somatic Embryogenesis Techniques; D018547:Plant Stems / Q000254:growth & development / Q000648:ultrastructure; D040901:Proteomics / Q000379:methods; D064028:Tracheophyta / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure
D010940:Plant Proteins
cryptogamic plants; hormone-free medium; mass spectrometry; transmission electron microscopy; two dimensional electrophoresis