Cytokinin (CK) content and activities of several antioxidant enzymes were examined during plant ontogeny with the aim to elucidate their role in delayed senescence of transgenic Pssu-ipt tobacco. Control Nicotiana tabacum L. (cv. Petit Havana SR1) and transgenic tobacco with the ipt gene under the control of the promoter of small subunit of Rubisco (Pssu-ipt) were both grown either as grafts on control rootstocks or as rooted plants. Both control plant types showed a decline in total content of CKs with proceeding plant senescence. Contrary to this both transgenic plant types exhibited at least ten times higher content of CKs than controls and a significant increase of CK contents throughout the ontogeny with maximal values in the later stages of plant development. Significantly higher portion of O-glucosides was found in both transgenic plant types compared to control ones. In transgenic plants, zeatin and zeatin riboside were predominant type of CKs. Generally, Pssu-ipt tobacco exhibited elevated activities of antioxidant enzymes compared to control tobacco particularly in the later stages of plant development. While in control tobacco activity of glutathione reductase (GR) and superoxide dismutase (SOD) showed increasing activity up to the onset of flowering and then gradually decreased, in both transgenic types GR increased and SOD activity showed only small change throughout the plant ontogeny. Ascorbate peroxidase (APOD) was stimulated in both transgenic types. The manifold enhancement of syringaldazine and guaiacol peroxidase activities was observed in transgenic grafts throughout plant ontogeny in contrast to control and transgenic rooted plants, where the increase was found only in the late stages. Electron microscopic examination showed higher number of crystallic cores in peroxisomes and abnormal interactions among organelles in transgenic tobacco in comparison with control plant. The overproduction of cytokinins resulted in the stimulation of activities of AOE throughout the plant ontogeny of transgenic Pssu-ipt tobacco.Additional key words: ascorbate peroxidase, catalase, glutathione reductase, guaiacol peroxidases, superoxide dismutase, syringaldazine peroxidase.⎯⎯⎯⎯ sequence of the gene coding for small subunit of Rubisco; SPOD -syringaldazine peroxidase; SOD -superoxide dismutase; T -transgenic rooted plants; T/C -transgenic grafts; TEM -transmission electron microscopy.
With the aim to contribute to the elucidation of the role of phytohormones in response of plants to adverse environmental conditions, seedlings of Phaseolus vulgaris, Nicotiana tabacum, Beta vulgaris, and Zea mays were supplied with water, 100 μM abscisic acid (ABA), or 10 μM N 6 -benzyladenine (BA) immediately before imposition of water stress (WS). In all four species, contents of chlorophylls (Chls) and carotenoids were markedly decreased during WS and after rehydration only in plants pre-treated with water but not in those pre-treated with ABA or BA. Contents of pigments of xanthophyll cycle increased during WS more in plants pre-treated with ABA or BA than in those pre-treated with water, but the degree of their de-epoxidation was highest in the later. Similarly, the efficiency of photosystem 2, determined as variable to maximal Chl fluorescence ratio, was not markedly decreased in bean plants pre-treated with ABA or BA in contrast to those pre-treated with water. The imposed WS was not severe enough to damage chloroplast ultrastructure. However, different changes in a size of starch inclusions were observed. In bean plants, the amount of starch increased considerably in plants pre-treated with water, while it decreased in BA pre-treated plants and no change was found in ABA pre-treated ones. The starch content declined under WS in sugar beet and tobacco plants but only moderate changes were found in ABA or BA pre-treated plants. Thus the application of BA and especially of ABA reduced the negative effects of subsequent WS.
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