Band 8, Heft 12

Plants are redox systems and redox-active compounds control and regulate all aspects of their life. Recent studies have shown that changes in reactive oxygen species (ROS) concentration mediated by enzymatic and non-enzymatic antioxidants are transferred into redox signals used by plants to activate various physiological responses. An overview of the main antioxidants and redox signaling in plant cells is presented. In this review, the biological effects of ROS and related redox signals are discussed in the context of acclimation to changing environmental conditions. Special attention is paid to the role of thiol/disulfide exchange via thioredoxins (Trxs), glutaredoxins (Grxs) and peroxiredoxins (Prxs) in the redox regulatory network. In plants, chloroplasts and mitochondria occupying a chloroplasts and mitochondria play key roles in cellular metabolism as well as in redox regulation and signaling. The integrated redox functions of these organelles are discussed with emphasis on the importance of the chloroplast and mitochondrion to the nucleus retrograde signaling in acclimatory and stress response.

The effect of a domain peptide DPCPVTc from the central region of the RYR2 on ryanodine receptors from rat heart has been examined in planar lipid bilayers. At a zero holding potential and at 8 mmol L−1 luminal Ca2+ concentration, DPCPVTc induced concentrationdependent activation of the ryanodine receptor that led up to 20-fold increase of PO at saturating DPCPVTc concentrations. DPCPVTc prolonged RyR2 openings and increased RyR2 opening frequency. At all peptide concentrations the channels displayed large variability in open probability, open time and frequency of openings. With increasing peptide concentration, the fraction of high open probability records increased together with their open time. The closed times of neither low- nor high-open probability records depended on peptide concentration. The concentration dependence of all gating parameters had EC50 of 20 μmol L−1 and a Hill slope of 2. Comparison of the effects of DPCPVTc with the effects of ATP and cytosolic Ca2+ suggests that activation does not involve luminal feed-through and is not caused by modulation of the cytosolic activation A-site. The data suggest that although “domain unzipping” by DPCPVTc occurs in both modes of RyR activity, it affects RyR gating only when the channel resides in the H-mode of activity.

Glucanase and chitinase enzymes play an important role in different plant processes including defense against pathogens and morphogenesis. Moreover, their role in the processes of somatic embryogenesis has been demonstrated. It has been suggested, that the presence of this type of proteins might be a marker for embryogenic potential of callus cultures. In this work we screened for the presence of glucanases and chitinases in liquid growth media of a set of conifer embryogenic cell lines in order to find correlation with their embryogenic potential. We have found that none of the 12 chitinase isoforms detected in culture media of Pinus nigra Arn. or the nine chitinases detected in media with Abies alba × A. cephalonica and Abies alba × A. numidica embryogenic tissues could be linked to their embryogenic capacity. Similarly, none of the six glucanase isoforms detected in the extracellular fluid of Pinus nigra Arn. cultures can be assigned as a marker of embryogenic potential. Thus, our data indicate the large variability and doubtless importance of glucanases and chitinases for cell growth and development of somatic embryos, however, do not support the premise that they are markers of embryogenesis.

Six allozymes of aspartate aminotransferase (AAT, EC 2.6.1.1): three plastidial (AAT-2 zone) and three cytosolic (AAT-3 zone) were isolated from common wheat (Triticum aestivum) seedlings and highly purified by a five-step purification procedure. The identity of the studied proteins was confirmed by mass spectrometry. The molecular weight of AAT allozymes determined by gel filtration was 72.4±3.6 kDa. The molecular weights of plastidial and cytosolic allozymes estimated by SDS-PAGE were 45.3 and 43.7 kDa, respectively. The apparent Michaelis constant (K m) values determined for four substrates appeared to be very similar for each allozyme. The values of the turnover number (k cat) and the k cat/K m ratio calculated for allozymes with L-aspartate as a leading substrate were in the range of 88.5–103.8 s−1/10,412–10,795 s−1 M−1 for AAT-2 zone and 4.6–7.0 s−1/527–700 s−1 M−1 for AAT-3 zone. These results clearly demonstrated much higher catalytic efficiency of AAT-2 allozymes. Therefore, partial sequences of cDNA encoding AATs from different zones were obtained using the RT-PCR technique. Comparison of the AAT-2 and AAT-3 amino acid sequences from active site regions revealed five non-conservative substitutions, which impact on the observed differences in the isozymes catalytic efficiency is discussed.

Seed germination of two local Greek endemics was studied (Erysimum naxense, Erysimum krendlii). Seed viability was determined by using the tetrazolium method and germination was studied in synchronized cycles of five and four alternating temperatures [10/5 (for E. naxense only) and 15/10, 20/15, 25/20, and 30/25°C for both species, in cycles of 16 h day/8 h night], and in five light regimes (red, blue, green, white, and dark). Germination of E. naxense and E. krendlii seeds was determined daily for six and five weeks, respectively, with the data analyzed as viability adjusted accumulative seed germination at the end of each week. E. naxense’s seed viability was higher (90%) than that of E. krendlii (64%); seed germination (%) of both increased at low alternating temperatures (10/5°C, 15/10°C, 20/15°C). Germination of E. naxense seeds at low temperatures was light-independent, whereas at high temperatures it was increased with red light. Germination of E. krendlii seeds was inconsistently affected by light at the temperatures studied. Percentages of seed germination of both species were higher in experimental conditions similar to the ones of their natural habitats during autumn and/or spring (facilitated with Geographic Information Systems). These conclusions provide guidelines for species-specific propagation protocols and ex situ conservation.

Inland saline habitats of the Pannonian Lowland exhibit a specific variety of grasslands determined by a soil salinity gradient. Changes in the hydrological regime and absence of management have resulted in heavy degradation of the vegetation. The impact of topsoil removal on salt steppes was tested by a 3-year small-scale manipulated experiment in SW Slovakia (Kamenínske Slanisko Nature Reserve). Topsoil was removed in three contrasting types of vegetation with different soil salinities, i.e. in different stages of habitat degradation. Data were analysed by multi-way ANOVA and by multivariate methods. Species richness decreased and the proportion of halophytes increased significantly in the two types with the highest soil salinity; however, the total number of halophytes was not influenced by soil removal. The treatment caused inhibition of secondary succession on the plots with the highest salinity. The effect of the soil removal was only short-term in the vegetation with moderate salinity and on heavily degraded and desalinized types it even stimulated further recruitment of ruderal species. Topsoil removal has only limited potential for the restoration of Pannonian salt steppes. It should be applied only in slightly degraded vegetation, where salt accumulation is still present and target species propagules are available.

Cyanobacteria are prokaryotic organisms characterized by their ability to produce secondary metabolites with different biological activities. The aim of this work was to evaluate the in vitro and in vivo toxicity of the cosmopolitan freshwater cyanobacterium H. kuetzingii. An extract from H. kuetzingii and cyanobacterial growth media were assessed for presence of intracellular and extracellular toxins by in vitro tests using primary cell cultures from mouse kidney and fibroblasts, cell lines A549 and 3T3, a fish cell line RTgill-W1 as well as by a traditional in vivo mouse bioassay. The presence of toxicity was compared with the ELISA and HPLC data for corresponding cyanotoxins. In vitro tests showed pronounced cytotoxicity of the cyanobacterium extract and growth medium in which H. kuetzingii released potential extracellular toxic compounds as the mammalian cells were significantly more sensitive to exposure compared to the fish cells. Histopathological analyses of the liver and kidneys of treated mice showed pathological changes such as leukocyte infiltration and necrosis, changes in the proximal and distal convoluted tubules, lack of differentiation of Bowman’s space, enlarged Bowman’s capsules and massive hemorrhages. ELISA and HPLC analyses confirmed the presence of saxitoxins and microcystins at low concentrations. In addition, the histological analyses suggest that H. kuetzingii produces other, yet unknown toxic metabolites. Monitoring efforts are therefore required to evaluate the potential hazard for the freshwater aquatic systems and possible public health implications associated with this cyanobacterium.

Anisomycin is a pyrrolidine antibiotic isolated from Streptomyces griseolus. It has been found that a quite low dose of anisomycin is sufficient to block proliferation of primary T lymphocytes. The focus of this study is to explore the possibility of anisomycin to treat human acute leukemia Jurkat T cells in vitro. The results indicated that the low dose of anisomycin could significantly inhibit the colony formation of Jurkat T cells and elevate the inhibition rate of Jurkat T cell growth along with its increasing concentrations. Jurkat T cell cycle was blocked into S-phase by anisomycin. Consistent with the increased proportion of sub-G1 phase, anisomycin promoted Jurkat T cell apoptosis. The CD69 and CD25 expression on the surface of Jurkat T cells was also down-regulated prominently along with the enhancing concentrations of anisomycin, followed by the decreased production of IL-4, IL-10, IL-17, TGF-β and IFN-γ, and the down-regulated expression of phosphorylated-ERK1/2. The results suggest that the suppressive effect of anisomycin on Jurkat T cell growth may be related to inhibiting TGF-β production and ERK1/2 activation, arresting the cell cycle at S-phase and promoting the apoptosis of Jurkat T cells.

This study examines the effect of irradiance level produced by solid-state light-emitting diodes (LEDs) on the growth, nutritional quality and antioxidant properties of Brassicaceae family microgreens. Kohlrabi (Brassica oleracea var. gongylodes, ‘Delicacy Purple’) mustard (Brassica juncea L., ‘Red Lion’), red pak choi (Brassica rapa var. chinensis, ‘Rubi F1’) and tatsoi (Brassica rapa var. rosularis) were grown using peat substrate in controlled-environment chambers until harvest time (10 days, 21/17°C, 16 h). A system of five lighting modules with 455, 638, 665 and 731 nm LEDs at a total photosynthetic photon flux densities (PPFD) of 545, 440, 330, 220 and 110 µmol m−2s−1 respectively were used. Insufficient levels of photosynthetically active photon flux (110 µmol m−2 s−1) suppressed normal growth and diminished the nutritional value of the Brassica microgreens studied. In general, the most suitable conditions for growth and nutritional quality of the microgreens was 330–440 µmol m−2 s−1 irradiation, which resulted in a larger leaf surface area, lower content of nitrates and higher total anthocyanins, total phenols and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging capacity. High light levels (545 µmol m−2 s−1), which was expected to induce mild photostress, had no significant positive impact for most of investigated parameters.

Research on moose-mediated seed dispersal is limited. However, its potential role in transferring seeds in patchy landscapes may be of great importance. In this work we examined how seasons and vegetation diversity influence the species richness and abundance of seeds dispersed endozoochorically by moose. Samples of moose faeces were collected year-round, fortnightly, from contrasting vegetation types, dominated by diverse, species-rich wetland or poor, dry pine forest. The viable seed content of dung was studied by the seedling emergence method. The mean number of emerged seedlings per 0.8 L sample and the mean number of plant species per 0.8 L sample were several times higher in the diverse wetland vegetation than in the poor pine forest vegetation. Maximum species richness and seed abundance was observed during the fructification period, and the minimum during spring. The species richness of samples did not differ between winter and the growing season, although the composition of plant species was different. The results of this study suggest that moose are efficient seed vectors, especially of grasses typical for grasslands and wetlands. The species richness and abundance of dispersed seeds coincides with the diversity of the vegetation of the animal’s habitat.

The abundance, composition and hydrocarbon-degrading bacteria, as possible biomarkers of contamination with oil hydrocarbons, of autochthonous and alochtonous microbiota of the digestive tract of rainbow trout have been estimated. The samples of the bottom sediments for microbiological tests have been collected and a response of natural bacterial communities in the digestive tract of rainbow trout and nutritional changes has been investigated. Experimental fish have been fed with a mixture of three substances with the aim to assess the influence of hydrocarbon-degrading bacteria contained in the sediments on the microbiota of rainbow trout’s digestive tracts. The abundance values of rainbow trout intestinal heterotrophic bacteria were found to change depending on alochtonous microbiota of different bottom sediments given to the experimental fish with food in vitro. According to the results of our research, it is likely that the changes in the abundance values of the microbiota of the digestive tract of fish and in the proportions of functional groups of the bacteria allow us to determine changes in the functional activity of bacteria depending on food composition. Any relative increase or decrease of abundance or activity of alochtonous microbiota allows the prediction of toxic effects of the contaminants on animals and the environment.

Paper by Li Li Huijuan Wu, Jialin Qian, Mingzhen Li, Yue Li, Baoming Li et al. “Decreased Na+/K+ ATPase α1 (ATP1A1) gene expression in major depression patients’ peripheral blood” in Voulme 8, Issue 11, 1077-1082 / November 2013; DOI: 10.2478/s11535-013-0207-8 contains incorrect units in Figure 3. The correct Figure 3, together with its caption is presented below.