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Changes in cytokinin pool and cytokinin oxidase/dehydrogenase activity (CKX EC: 1.5.99.12) in response to increasing abscisic acid (ABA) concentrations (0.5–10 μM) were assessed in the last fully expanded leaves and secondary roots of two pea (Pisum sativum) varieties with different vegetation periods. Certain organ diversity in CKX response to exogenous ABA was observed. Treatment provoked altered cytokinin pool in the aboveground parts of both studied cultivars. Specific CKX activity was influenced significantly basically in roots of the treated plants. Results suggest that ABA-mediated cytokinin pool changes are leaf-specific and involve certain root signals in which CKX activity presents an important link. This enzymatic activity most probably regulates vascular transport of active cytokinins from roots to shoots.
The aim of the experiments presented here was twofold. On the one hand, to study the influence of plant growth regulators, i.e. auxins (IAA and NAA), cytokinins (kinetin, zeatin and zeatin riboside), gibberellins: (GA₃) and polyamines (spermidine, spermine and putrescine) on the generative development of winter wheat plants, while on the other to study the interaction of plant growth regulators with electric current in affecting the efficiency of the flowering of wheat plants. Winter wheat var. Grana was subjected to a short vernalization (2 weeks at 5℃ + 10 days at 10℃) on Murashige and Skoog medium containing growth regulators at different concentrations. For selected substances, seedlings were additionally treated with electric current by applying a constant voltage for a given period of time (30 V for 30 s or 1.5 V for 1 h) with the anode or the cathode inserted into apical leaves and the reference electrode into the media. After that, the seedlings were transferred to a glasshouse (20/17℃) where they were grown until heading was achieved. All the substances studied stimulated the generative development of winter wheat, in contrast to non-treated plants (control), especially at their highest concentrations. Spermidine, kinetin and GA₃ were the most effective in this process as they stimulated 100–80% of generative plants (whereas for the control the percentage stood at only 30% plants). The studied regulators also increased the rate of generative development, i.e. shortening the length of the vegetative phase (by about 30% in comparison with the control). Moreover, these substances treatments decreased the fresh mass of both seedlings and flowering plants and increased the number of spikelets in the ear. The electric current treatment interacted with the applied substances: the anode insertion into the leaves generally increased, whereas the cathode insertion decreased, the inductive effect of the growth regulators on generative development. However, the inhibition of the flowering process was more pronounced than the induction. Moreover, the passage of electric current shortened the time to heading. This decrease in the length of the vegetative phase was more visible after the anode was inserted into the leaves. The least noticeable effect of the electric current interaction with plant growth regulators on both the generative development and the time of heading was observed for auxins.
Vernalization-induced flowering is an effect of the epigenetic regulation of gene expression through DNA methylation and histone modifications. Vernalizationmediated silencing of a floral repressor through histone modifications was shown in Arabidopsis thaliana. However, for Brassica napus L., the mechanism underlying vernalization is unclear, and the roles of DNA methylation and histone modifications have not been established. This study revealed the profiles of changes in the DNA methylation state during vernalization (after 14, 35, 56 days) and the subsequent growth in long- or short-day photoperiods (after 2, 7, 14 days) in the winter and spring rapeseed using TLC and MSAP techniques. TLC analysis showed a significant decrease in the amount of 5-methylcytosine (m5C) in genomic DNA in both cultivars at the beginning of vernalization, but upon its termination, the winter rape showed a reduced level of m5C contrary to a significantly increased level in the spring rape. MSAP analysis revealed that winter and spring rapeseed differed in the MSAP loci which were demethylated/methylated in the course of the experiment and presented diverse profiles of changes in the methylation state. The winter rape showed permanent demethylations at 69.2 % of MSAP loci in the course of vernalization that were mostly preserved upon its termination. The spring rape showed similar numbers of demethylations and methylations that were mainly transient. The study provides evidence of the role of DNA methylation in vernalization for rapeseed and for the significant prevalence of demethylations at the beginning of vernalization, which is necessary for the transition to reproductive growth.
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