Abstract
BackgroundSomatic embryogenesis (SE) is a complex biological process that occurs under inductive conditions and causes fully differentiated cells to be reprogrammed to an embryo like state. In order to get a better insight about molecular basis of the SE in Crocus sativus L. and to characterize differentially accumulated proteins during the process, a proteomic study based on two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry has been carried out.ResultsWe have compared proteome profiles of non-embryogenic and embryogenic calli with native corm explants. Total soluble proteins were phenol-extracted and loaded on 18 cm IPG strips for the first dimension and 11.5% sodium dodecyl sulfate-polyacrylamide gels for the second dimension. Fifty spots with more than 1.5-fold change in abundance were subjected to mass spectrometry analysis for further characterization. Among them 36 proteins could be identified, which are classified into defense and stress response, protein synthesis and processing, carbohydrate and energy metabolism, secondary metabolism, and nitrogen metabolism.ConclusionOur results showed that diverse cellular and molecular processes were affected during somatic to embryogenic transition. Differential proteomic analysis suggests a key role for ascorbate metabolism during early stage of SE, and points to the possible role of ascorbate-glutathione cycle in establishing somatic embryos.
Highlights
Somatic embryogenesis (SE) is a complex biological process that occurs under inductive conditions and causes fully differentiated cells to be reprogrammed to an embryo like state
We found that the proteome profiles of nodular calli (NC) and non-embryogenic calli (NEC) were significantly different compared to the original corm explants, which specifies complete reprogramming of gene expression taking place to support somatic to embryogenic transition
RNA binding protein 45 (RBP45) increased significantly in non-embryogenic and embryogenic calli. In conclusion, this is the first proteomics analysis that examines the proteomic changes that occur during induction of SE in saffron. two-dimensional gel electrophoresis (2-DE) combined to mass spectrometry led to the identification of several different functional categories of proteins that might be involved in SE
Summary
Somatic embryogenesis (SE) is a complex biological process that occurs under inductive conditions and causes fully differentiated cells to be reprogrammed to an embryo like state. Saffron (Crocus sativus L., Iridaceae) has long been cultivated for the production of saffron spice, which makes it interesting from an economic as well as a scientific point of view. Plant growth regulators (PGRs) have a critical role in SE induction and subsequent modulation of the proper morphogenesis in embryo development. Decreasing or removal of exogenous auxin is necessary for embryo morphogenesis and further development [5]. During SE, differentiated somatic cells undergo a series of morphological and biochemical changes and are completely reprogrammed to an embryonic like state which forms the basis of cellular totipotency in plants [8]. Understanding the molecular and biochemical pathways that initiate and direct vegetative to embryogenic transition is of great importance to plant molecular biologists
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