To gain a better understanding of cold acclimation process in wheat, we applied a 2-DE based proteomic approach to discover changes in proteome profile of a diploid wild wheat (Triticum urartu L.) during prolonged cold stress treatment. To this end, plants were grown in pots and the growing seedlings (4-leaf stage) were exposed to cold stress. After 4 weeks of cold acclimation (4-6 °C) and subsequent treatment for 12 h at -2 °C, samples were collected from control and stressed plants and were subjected to proteome pattern analysis. Among approximately 450 reproducible protein spots displayed in each given 2-DE gels, 34 proteins changed significantly in abundance in response to cold stress. Among them, 25 and 9 proteins were up and down-regulated under stress condition, respectively. Analysis by matrix-assisted laser desorption ionization time of flight/time of flight mass spectrometry coupled with non-redundant protein database search allowed the identification of 20 cold-induced proteins. Integrated proteomic and database survey resulted in identification of several cold stress related proteins such as pathogenesis related protein, cold regulated protein, cold-responsive LEA/RAB-related COR protein, oxygen-evolving enhancer protein and oxalate oxidase. The presumed functions of the identified proteins were mostly related to cold acclimation, oxidative stress and photosynthesis. The possible implications of differentially accumulated proteins in acquiring systemic tolerance to freezing stress following exposure to prolonged low temperature will be discussed.
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.
Saffron (Crocus sativus L.) is a monocotyledonous plant propagated via corms, but recently several alternative methods have been reported. To find the conditions suitable for saffron shoot formation from corms, the effect of different concentrations of the plant growth regulatory cytokinins N-6-benzyladenine (BA) and N-phenyl-1, 2,3-thidiazol-5-ylurea, commonly known as thidiazuron (TDZ), were compared. In all corm explants, an average of 39.5 +/- A 5.1 shoots per corm were induced by 4.54 mu M TDZ, whereas only 3.6-11.4% by BA. The outstanding result in the shoot formation stage is the generation of globular, translucent structures that are morphologically similar to globular embryos. To optimize the plant regeneration from the induced adventitious shoots obtained from the TDZ treatment, the shoots were transferred to MS and B5 media supplemented with different concentrations and combinations of NAA and BA. The highest rate of plant regeneration from developing shoots was observed in the B5 medium containing 2.22 mu M NAA and 2.68 mu M BA. With optimized hormonal conditions, an average of 19.55 +/- A 5.75 shoots and 3.18 +/- A 1.5 roots per explants were obtained. Based on this experiment, a simple, new and efficient protocol is presented to produce numerous plants from induced corm explants of saffron
Among the different concentrations of Thidiazuron (TDZ) and between the two media Gamborg (B5) and Murashige and Skoog (MS), the highest frequency of shoot formation could be seen in the MS medium with TDZ concentration of 4.54 microM. Among the different concentrations of Naphtalene acetic acid (NAA) and Benzyl adenine (BA) in the two aforementioned media, the maximum proliferation and rooting of saffron shoots were obtained in a B5 medium containing 2.22 microM NAA and 2.68 microM BA. Peroxidase (POD), catalase (CAT), superoxide dismutase (SOD), esterase (EST) and polyphenoloxidase (PPO) measurements proved that all the enzymes had a similar pattern of changes, according to which their concentrations increased in the first stages of development and then decreased. The same pattern was observed for polyphenoloxidase in a B5 medium while in the MS medium a reverse pattern was observed. The enzyme concentration decreased and then increased during shoot formation. The results show the principal role of antioxidant enzymes in the complicated process of organogenesis.
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