Transcription Factors of the GLRs Family Are Involved in Cytokinin-Dependent Regulation of Plastid RNA Polymerase SCA3 Gene Expression during Deetiolation of Arabidopsis thaliana.

Light-dependent transcription factors GLKs of Arabidopsis thaliana are involved in the anterograde regulation of chloroplast biogenesis during deetiolation: they regulate the expression of photosynthetic nuclear-encoded genes and also mediate the transcription of plastid genes. Chloroplast biogenesis is determined at the same time by light and by endogenous factors (phytohormones), among which cytokinins significantly accelerate the formation of photosynthetically active chloroplasts. In this work, it was shown that trans-factors GLKs function as cytokinin-dependent regulators, mediating the positive cytokinin effect on the plastome expression through the activation of transcription of the SCA3 nuclear gene encoding the plastid RNA polymerase RPOTp.

The Melatonin Receptor CAND2/PMTR1 Is Involved in the Regulation of Mitochondrial Gene Expression under Photooxidative Stress.

Melatonin is a signaling molecule that mediates multiple stress-dependent reactions. Under photooxidative stress conditions generating intensive ROS production, exogenous melatonin (50 µM) contributed to maintaining the expression of mitochondrial encoded genes and up-regulation of RNA-polymerase genes RPOTm and RPOTmp, operating through the CAND2 receptor and α-subunit of the heterotrimeric G protein GPA1 coupled with CAND2. Unlike wild-type plants, mutants with defective CAND2 and GPA1 genes exhibited no decrease in the alternative pathway of leaf respiration, as well as the activity of an alternative oxidase, and the expression of the AOX1a gene. At the same time, the protective effect of exogenous melatonin on some physiological indicators did not depend on the receptor and was associated with the direct antioxidant function of the regulator. Thus, melatonin under photooxidative stress conditions can act as an antioxidant and as a hormone capable of regulating the expression of nuclear and organelle genes through the components of melatonin signal perception.

Ontogenetic, Light, and Circadian Regulation of PAP Protein Genes during Seed Germination of Arabidopsis thaliana.

The expression profiles of the PAP genes, encoding proteins associated with plastid multisubunit RNA polymerase, were studied in dry seeds, during germination, and at the early stages of Arabidopsis thaliana seedling formation. A detailed analysis of the PAP transcript levels by RT-PCR showed that the transition of seeds from dormancy to active growth is accompanied by a drastic increase in the transcript accumulation of all studied genes on the first day of germination, both in the light and in the dark. Further changes in transcript levels differed among PAP genes and were apparently determined by their functional specificity. It was established for the first time that the expression of individual PAP genes is regulated by circadian rhythms, in addition to factors of ontogenetic and light nature.

GRUSP, an Universal Stress Protein, Is Involved in Gibberellin-dependent Induction of Flowering in Arabidopsis thaliana.

The effect of T-DNA insertion in the 3'-UTR region of Arabidopsis thaliana At3g58450 gene encoding the Germination-Related Universal Stress Protein (GRUSP) was studied. It was found that under a long-day condition this mutation delays transition to flowering of grusp-115 transgenic line that due to a reduced content of endogenous bioactive gibberellins GA1 and GA3 in comparison to the wild-type plants (Col-0). Exogenous GA accelerated flowering of both lines but did not change the time of difference in the onset of flowering between Col-0 and grusp-115. In addition to changes in GA metabolism, grusp-115 evidently has disturbances in realization of the signal that induces flowering. This is confirmed by the results of gene expression of the floral integrator FLOWERING LOCUS T (FT) and the floral repressor FLOWERING LOCUS C (FLC), which are key flowering regulators and acting opposite. We hypothesize that the formation of grusp-115 phenotype can also be affected by a low expression level of FT due to up-regulated FLC expression.

Cold Stress Activates the Expression of Genes of the Chloroplast Transcription Apparatus in Arabidopsis thaliana Plants.

The physiological and molecular responses of Arabidopsis thaliana plants to cold stress were studied. Exposure to a low non-freezing temperature (4°C, 5 days) caused a decrease in the physiological functions and activity of a number of photosynthetic genes and elevation in expression of the cold stress gene COR15a, the product of which protects chloroplasts. It was shown for the first time that in parallel to a general inhibition of physiological functions under hypothermia, an increase in the expression of most genes for the chloroplast transcription apparatus was observed. This is obviously one of the compensatory mechanisms of adaptation aimed to maintain cellular homeostasis and physiological functions under hypothermia.

The Transcription Factor HY5 Is Involved in the Cytokinin-Dependent Regulation of the Expression of Genes Encoding Proteins Associated with Bacterial Plastid RNA-Polymerase during De-Etiolation of Arabidopsis thaliana.

HY5 (ELONGATED HYPOCOTYL5), a bZIP transcription factor, is one of the main regulators of light and hormonal signaling. Among the targets of this gene, the genes for the transcriptional complex of chloroplasts whose coordinated expression ensures the initial stages of photomorphogenesis are particularly significant. In this study, we showed that, during de-etiolation, HY5 functions as a positive CK-dependent regulator of the expression of genes encoding proteins associated with plastid RNA polymerase (PAP), which functions below the primary chain of sensing the cytokinin signal. The absence of blocking effect of mutations of the CRY1, CRY2, PHYA, and PHYB photoreceptor genes on the CK-dependent content of PAP gene transcripts indicates the parallel action of the hormone and light in their regulation.

Melatonin Inhibits Peroxide Production in Plant Mitochondria.

The effect of melatonin on respiration and production (release) of hydrogen peroxide during succinate oxidation in mitochondria isolated from lupine cotyledons and epicotyls of pea seedlings was studied. It was shown for the first time that melatonin (10-7-10-3 M) had a significant inhibitory effect on the production of peroxide by plant mitochondria, which was characterized by concentration dependence and species specificity. At the same time, melatonin (at a concentration of up to 100 µM) had virtually no effect on mitochondrial respiration rate and respiratory control coefficient. The results confirm the antioxidant function of melatonin and indicate that it is involved in the regulation of ROS levels and maintenance of redox balance in plant mitochondria.

Detection of Taxuyunnanin C in Suspension Cell Culture of Taxuscanadensis.

This is the first study to isolate the taxoid taxuyunnanin C (group of 14-hydroxylated taxoids) from the biomass of suspension cell culture of the Canadian yew (Taxus canadensis). According to available data, this is the first report of the presence of nonpolar (polyacylated) forms of 14-hydroxylated taxoids, including taxuyunnanin C, in T. canadensis.

The effect of light quality on the pro-/antioxidant balance, activity of photosystem II, and expression of light-dependent genes in Eutrema salsugineum callus cells.

The antioxidant balance, photochemical activity of photosystem II (PSII), and photosynthetic pigment content, as well as the expression of genes involved in the light signalling of callus lines of Eutrema salsugineum plants (earlier Thellungiella salsuginea) under different spectral light compositions were studied. Growth of callus in red light (RL, maximum 660 nm), in contrast to blue light (BL, maximum 450 nm), resulted in a lower H2O2 content and thiobarbituric acid reactive substances (TBARS). The BL increased the activities of key antioxidant enzymes in comparison with the white light (WL) and RL and demonstrated the minimum level of PSII photochemical activity. The activities of catalase (CAT) and peroxidase (POD) had the highest values in BL, which, along with the increased H2O2 and TBARS content, indicate a higher level of oxidative stress in the cells. The expression levels of the main chloroplast protein genes of PSII (PSBA and PSBD), the NADPH-dependent oxidase gene of the plasma membrane (RbohD), the protochlorophyllide oxidoreductase genes (POR B, C) involved in the biosynthesis of chlorophyll, and the key photoreceptor signalling genes (CIB1, CRY2, PhyB, PhyA, and PIF3) were determined. Possible mechanisms of light quality effects on the physiological parameters of callus cells are discussed.

Phytohormones Regulate the Expression of Nuclear Genes Encoding the Components of the Plastid Transcription Apparatus.

As multifunctional regulators of physiological processes, phytohormones play an important role in the regulation of expression of the plastid genome and chloroplast biogenesis. Hormones can directly regulate the expression of genes localized in the chloroplast genome. However, many components of the plastid transcription apparatus are encoded by nuclear rather than plastid genes. It remains obscure whether these nuclear genes are subject to hormonal regulation. This is the first study to show that phytohormones exert differential effects on the expression of nuclear genes of the transcription machinery of the Arabidopsis thaliana plastome. RT-PCR analysis showed that the level of transcripts of the majority of studied genes was activated by trans-zeatin but decreased under the influence of ABA, methyl jasmonate, and salicylic acid, whereas ethylene had no significant effect, and the effects of brassinolide depended on the illumination conditions. The results of this study indicate that the hormonal regulation of the plastome expression can be mediated by differential regulation of the nuclear genes encoding plastid transcription machinery components.

The Gene Encoding the Universal Stress Protein AtUSP is Regulated by Phytohormones and Involved in Seed Germination of Arabidopsis thaliana.

For the first time, the organ-specific expression pattern of the AtUSP (At3g58450) gene, which also undergoes hormonal regulation, was shown. The USP protein encoded by this gene is involved in seed germination of Arabidopsis thaliana and, unlike abscisic acid, stimulates this process.

Occurrence of 14-hydroxylated taxoids in the plant in vitro cell cultures of different yew species (Taxus spp.).

This is the first study to show that the formation of 14ß-hydroxylated derivatives of taxa-4(20),11-diene is a specific feature of in vitro cultured dedifferentiated yew cells that distinguishes them from intact plant cells. This may be due to a lower toxicity of the 14-OH taxoids for proliferating plant cells compared to the 13-OH derivatives.

Brassica napus responses to short-term excessive copper treatment with decrease of photosynthetic pigments, differential expression of heavy metal homeostasis genes including activation of gene NRAMP4 involved in photosystem II stabilization.

In the present study, the influence of 50 and 100 µM CuSO4 was investigated starting from 3 h till 72 h treatment of 4-weeks Brassica napus plants. High CuSO4 concentrations in nutrient medium resulted in the rapid copper accumulation in plants, especially in roots, much slower and to lower degree in leaves. Copper excess induced early decrease in the leaf water content and temporary leaf wilting. The decrease in content of photosynthetic pigments became significant to 24 h of excessive copper treatments and reached 35 % decrease to 72 h, but there were no significant changes in maximum quantum efficiency of photosystem II photochemistry. The copper excess affected the expression of ten genes involved in heavy metal homeostasis and copper detoxification. The results showed the differential and organ-specific expression of most genes. The potential roles of copper-activated genes encoding heavy metal transporters (ZIP5, NRAMP4, YSL2, and MRP1), metallothioneins (MT1a and MT2b), low-molecular chelator synthesis enzymes (PCS1 and NAS2), and metallochaperones (CCS and HIPP06) in heavy metal homeostasis and copper ion detoxification were discussed. The highest increase in gene expression was shown for NRAMP4 in leaves in spite of relatively moderate Cu accumulation there. The opinion was advanced that the NRAMP4 activation can be considered among the early reactions in the defense of the photosystem II against copper excess.

Effect of red light on photosynthetic acclimation and the gene expression of certain light signalling components involved in the microRNA biogenesis in the extremophile Eutrema salsugineum.

The photosynthetic acclimation of extremophile Eutrema salsugineum plants to red light (RL) (14 days, 150 µmol photons m-2 s-1, 660 nm) and the expression of the key photoreceptor apoprotein genes, transcription factors (TFs) and associated with phytochrome system MIR (microRNA) genes were studied. RL exposure induced an increase in the content of anthocyanin and total phenolic compounds and the level of Chls was decreased. The photosystem 2 electron transport rate and the number of open reaction centres (qL) were not changed in RL plants, however, the levels of non-photochemical quenching (NPQ) and the regulated quantum yield of non-photochemical quenching Y(NPQ) were significantly higher in the RL plants. The rate of CO2 uptake was decreased by almost 1.4-fold but the respiration and transpiration rates, as well as the stomatal conductance were not changed in the RL plants. An increase in the expression of the photoreceptor apoprotein genes PHYA, PHYB and PHYC, the TF genes PIF4, PIF5 and miR395, miR408, miR165 and decreases in the levels of the transcripts of the TF gene HY5 and miR171, miR157, and miR827 were detected. The acclimation effect of photosynthetic apparatus to RL was accompanied by an increase of pigment content such as total phenolic compounds and carotenoids and it is due to the changes in the expression of the apoprotein phytochrome genes PHYA, PHYB, PHYC and phytochrome signalling TFs (PIF4, PIF5 and HY5) as well as MIR genes associated with phytochrome system.

Nickel accumulation in rape shoots (Brassica napus L.) increased by putrescine.

Application of exogenous putrescine (Put) increases nickel accumulation in rape shoots (Brassica napus), improving potential for phytoremediation of contaminated soils. Plants were grown within a growth chamber in water culture for five weeks, then 250-500 microM NiCl2 was added to rooting media. Within 5 days of treatment, damaging effects of nickel manifested as a reduction of root system size and a decrease of Cu and especially Fe content in young leaves. Spraying leaves of adult plants with Put markedly reduced toxic effects of Ni on root growth, enhanced leaf supply with Fe, and increased Ni content in young leaves by 2.5 times. Plant growth in medium with elevated levels of Ni stimulated accumulation of endogenous spermidine (Spd), spermine (Spm), and especially Put (by 4 times as compared with the control). Results suggest that Ni-induced accumulation of endogenous polyamines in rape leaves is caused by activation of long-distance metal transport within the plant and reduction of its toxicity due to Put chelating action.