Azamat Avalbaev

Cytokinin Stimulates Chloroplast Transcription in Detached Barley Leaves

Chloroplasts are among the main targets of cytokinin action in the plant cell. We report here on the activation of transcription by cytokinin as detected by run-on assays with chloroplasts isolated from apical parts of first leaves detached from 9-d-old barley (Hordeum vulgare) seedlings and incubated for 3 h on a 2.2 × 10−5 m solution of benzyladenine (BA). Northern-blot analysis also detected a BA-induced increase in the accumulation of chloroplast mRNAs. A prerequisite for BA activation of chloroplast transcription was preincubation of leaves for 24 h on water in the light, resulting in a decreased chloroplast transcription and a drastic accumulation of abscisic acid. Cytokinin enhanced the transcription of several chloroplast genes above the initial level measured before BA treatment, and in the case of rrn16 and petD even before preincubation. Cytokinin effects on basal (youngest), middle, and apical (oldest) segments of primary leaves detached from plants of different ages revealed an age dependence of chloroplast gene response to BA. BA-induced stimulation of transcription of rrn16, rrn23, rps4, rps16, rbcL, atpB, and ndhC required light during the period of preincubation and was further enhanced by light during the incubation on BA, whereas activation of transcription of trnEY, rps14, rpl16, matK, petD, and petLG depended on light during both periods. Our data reveal positive and differential effects of cytokinin on the transcription of chloroplast genes that were dependent on light and on the age (developmental stage) of cells and leaves.

Stimulation of wheat germ agglutinin gene expression in root seedlings by 24-epibrassinolide

The effects of 24-epibrassinolide (EB) on the growth of four-day-old seedlings of Triticum aestivum L. roots, the accumulation of wheat germ agglutinin (WGA) in them, and its gene expression were studied. EB stimulated growth with two optimum concentrations, 0.4 nM and 0.4 μM. Both concentrations enhanced WGA accumulation in roots, which resulted from the stimulation of its gene expression. The latter was shown by dot-blot hybridization of transcripts with a DNA probe corresponding to WGA. Since EB did not affect the level of endogenous ABA, its direct effect on WGA-gene transcription in wheat roots is suggested.

Wheat germ agglutinin regulates cell division in wheat seedling roots

The mitogenic activity of wheat germ agglutinin (WGA) has been studied in roots of 4-day-old wheat seedlings. WGA had a more pronounced stimulating effect on cell division than the known mitogens concanavalin A and phytohemagglutinin whereas gliadin had no effect. Treatment of wheat seedling roots with exogenous WGA led to the accumulation of indoleacetic acid and cytokinins, hormones that play an important role in the activation of plant cell growth. The data on the combined effect of 24-epibrassinolide and WGA on cell division and accumulation of phytohormones in seedling roots support a possible link between the endogenous WGA level and hormonal regulation of cell division in the root meristem of wheat plants.

Cytokinin oxidase is involved in the regulation of cytokinin content by 24-epibrassinolide in wheat seedlings

Fast and stable 2-fold accumulation of cytokinins (CKs) was detected initially in roots and then in shoots of 4-day-old wheat (Triticum aestivum L.) seedlings in the course of their treatment with 0.4μM 24-epibrassinolide (EBR). Elevated cytokinin level has been maintained only in the presence of EBR, while the hormone removal has led to return of cytokinin concentration to the control level initially in the roots and then in the shoots. EBR-induced accumulation of cytokinins was accompanied by inhibition of both cytokinin oxidase (CKX) (cytokinin oxidase/dehydrogenase, EC 1.5.99.12) activity and expression of the gene coding for this enzyme, and on the contrary the decline in CKs level resulted in increase in these characteristics up to the control level in roots and then in shoots. Sharp accumulation of cytokinin O-glucosides has been discovered in response to EBR-treatment suggesting fast EBR-induced activation of production of cytokinins, which excessive amounts were transferred into the storage forms. The obtained data provide evidence for the involvement of EBR in regulation of cytokinin level in wheat seedlings.

Induction of wheat germ agglutinin synthesis by abscisic and gibberellic acids in roots of wheat seedlings

The dynamics of ABA and wheat germ agglutinin (WGA) content and WGA-geneexpression have been studied in roots of 4-d-old wheat seedlings treated with 4 ABA and 0.3 GA. Both hormones caused rapidand significant effects on WGA accumulation, though GA might control thisprocess independently of ABA, since the content of endogenous ABA did not changeafter GA-treatment. The results of dot-blot analysis of transcripts of RNA usingthe 32P labelled WGA-gene as a probe showed that a two-fold increasein WGA content resulted from ABA- and GA-induced activation of WGA-geneexpression.

Cytokinin oxidase is key enzyme of cytokinin degradation.

Cytokinin oxidase (EC 1.5.99.12) is an enzyme that catalyzes the irreversible degradation of cytokinin phytohormones that are extremely necessary for growth, development, and differentiation of plants. Cytokinin oxidase plays an important role in the regulation of quantitative level of cytokinins and their distribution in plant tissues. This review generalizes the available information on the structure, properties, and functional role of this enzyme in plant ontogeny under conditions of normal growth and under the influence of unfavorable environmental factors.

Control Mechanisms of Lectin Accumulation in Wheat Seedlings under Salinity

Using the inhibitory analysis, we demonstrated that 0.3 M NaCl and 3.7 μM ABA induced wheat germ agglutinin (WGA) accumulation in 4-day-old wheat (Triticum aestivum) seedlings under conditions of suppressed transcription and translation owing to some rapid mechanisms controlling the level of this protein. Exogenous WGA could protect wheat seedlings against salinity by preventing the salt-induced inhibition of mitotic activity in the apical root meristem.

Expression profiles and hormonal regulation of tobacco expansin genes and their involvement in abiotic stress response

Changes in the expression levels of tobacco expansin genes NtEXPA1, NtEXPA4, NtEXPA5, and NtEXPA6 were studied in different organs of tobacco (Nicotiana tabacum L.) as well as in response to phytohormone and stress treatments. It was shown that NtEXPA1, NtEXPA4 and NtEXPA5 transcripts were predominantly expressed in the shoot apices and young leaves, but almost absent in mature leaves and roots. The NtEXPA6 mRNA was found at high levels in calluses containing a large number of undifferentiated cells, but hardly detectable in the leaves of different ages and roots. In young leaves, expression levels of NtEXPA1, NtEXPA4 and NtEXPA5 genes were induced by cytokinins, auxins and gibberellins. Cytokinins and auxins were also found to increase NtEXPA6 transcripts in young leaves but to the much lower levels than the other expansin mRNAs. Expression analysis demonstrated that brassinosteroid phytohormones were able either to up-regulate or to down-regulate expression of different expansins in leaves of different ages. Furthermore, transcript levels of NtEXPA1, NtEXPA4, and NtEXPA5 genes were increased in response to NaCl, drought, cold, heat, and 10μM abscisic acid (ABA) treatments but reduced in response to more severe stresses, i.e. cadmium, freezing, and 100μM ABA. In contrast, no substantial changes were found in NtEXPA6 transcript level after all stress treatments. In addition, we examined the involvement of tobacco expansins in the regulation of abiotic stress tolerance by transgenic approaches. Transgenic tobacco plants with constitutive expression of NtEXPA1 and NtEXPA5 exhibited improved tolerance to salt stress: these plants showed higher growth indices after NaCl treatment and minimized water loss by reducing stomatal density. In contrast, NtEXPA4-silenced plants were characterized by a considerable growth reduction under salinity and enhanced water loss. Our findings indicate that expression levels of all studied tobacco expansins genes are modulated by plant hormones whereas NtEXPA1, NtEXPA4, and NtEXPA5 expansins may be involved in the regulation of stress tolerance in tobacco plants.

Influence of 24-epibrassinolide on growth of wheat plants and the content of dehydrins under cadmium stress

Effect of pretreatment of wheat (Triticum aestivum L.) seedlings with 0.4 μM 24-epibrassinolide (EB) on subsequent action on them of 1 mM cadmium acetate was investigated. This pretreatment decreased the extent of damage of cadmium action on wheat plants as judged by inhibition of stress-induced efflux of electrolytes from plant tissues and plant ability to maintain the growing parameters at a level close to the control. It was established that the revealed ability of EB to induce twofold increased accumulation of 28 kDa dehydrin under normal plant growth conditions and even to cause additional accumulation of this protein under the cadmium stress is of importance in development of the preadapting and protection effects of EB on wheat seedlings. Using fluridone (FD), a known inhibitor of the biosynthesis of ABA, the pathways responsible for the regulation of 28 kDa dehydrin level and both dependent and independent on ABA were revealed in wheat plants both pretreated and untreated with EB and under cadmium stress.

Role of AINTEGUMENTA-like gene NtANTL in the regulation of tobacco organ growth

The Nicotiana tabacum AINTEGUMENTA-like gene (NtANTL), encoding one of AP2/ERF transcription factors, is a putative ortholog of the AtANT gene from Arabidopsis thaliana. In wild-type tobacco plants, the NtANTL gene was expressed in the actively dividing young flowers, shoot apices, and calluses, while the level of its mRNA increased considerably after treatment with exogenous 6-benzylaminopurine, indoleacetic acid and 24-epibrassinolide. We found a positive correlation among the expression levels of NtANTL, cyclin NtCYCD3;1 and cyclin-dependent kinase NtCDKB1-1 genes, suggesting possible molecular links between AINTEGUMENTA and cell cycle regulators in tobacco plants. However, no correlation was observed between NtANTL, NtCYCD3;1 and NtCDKB1-1 expression levels in response to NaCl and ABA. These observations indicate that the transcription factor NtANTL was not involved in the regulation of the cellular response to salinity nor did it affect the expression of NtCYCD3;1 and NtCDKB1-1 when tobacco plants were exposed to salt stress and ABA. In addition, we generated transgenic tobacco plants with both up-regulated and down-regulated expression of the NtANTL gene. Constitutive expression of the NtANTL gene contributed to an increase in the size of leaves and corolla of transgenic plants. Transgenic plants with reduced expression of the NtANTL gene had smaller leaves, flowers and stems, but showed a compensatory increase in the cell size of leaves and flowers. The results show the significance of the NtANTL gene for the control of organ growth by both cell division and expansion in tobacco plants.