Cytokines In Formation Research Articles

The content of endogenous hormones in explants and calluses of <i>Lavandula angustifolia</i> Mill. at the initial stages of <i>in vitro</i> culture

The content of endogenous hormones (auxin IAA, cytokinines, ABA) in explants of various types (segments of leaf, bud, stem), primary calluses induced from them, as well as morphogenic and non-morphogenic calluses at the initial stages of in vitro culture by the immunoassay method was studied for the first time for Lavandula angustifolia Mill. The maximum value of hormone levels in such explants as segments of bud was shown. An increase in the content of hormones in primary calluses was revealed in comparison with similar characteristics in all types of explants. The higher level of the active form of cytokinin (trans-zeatin) and ABA, as well as the lower level of the inactive form of cytokinin (zeatin-nucleotide) and auxin IAA were identified in morphogenic callus compared with non-morphogenic callus. It is suggested that the content of endogenous hormones in explants and calluses of L. angustifolia is due to their histological status. The conclusion is made about the unified histophysiological mechanisms of callusogenesis and morphogenesis in vitro in the studied plant.

Development and validation of a model for early survival prediction following liver transplantation based on donor and recipient characteristics

Background Circulating cytokine levels not only correlate with the progression of liver disease but also serve as indicators for the infection status of the body. Growing evidence points to the connection between donor cytokines and graft function following transplantation. This study set out to explore the clinical significance of donor cytokines in predicting liver transplantation prognosis. Methods Data from 172 deceased donor liver transplantations conducted between 2017 and 2022, with available donor serum cytokine information, were collected. The subjects were randomly divided into estimation (n = 120) and validation (n = 52) groups to establish and validate the model. The newly developed SA10 score was compared against established models EAD, MEAF, L-GrAFT7, and L-GrAFT10. Results Donor IL-10, along with donor age and recipient AST peak value within the first 7 days post-operation, was identified as an independent factor associated with recipient survival and was incorporated into the SA10 score. SA10 exhibited robust predictive capability, particularly for 1-month survival (AUC = 0.90, 95% CI = 0.84–0.96), outperforming EAD (AUC = 0.75, 95% CI = 0.60–0.90, p = 0.04) and L-GrAFT7 (AUC = 0.65, 95% CI = 0.49–0.81, p < 0.01). Comparable performance was observed between SA10, MEAF, and L-GrAFT10. Conclusion Donor IL-10 independently influences recipient survival, with the SA10 score demonstrating comparable and even superior predictive ability compared to existing models.

Hormonal profiles in dormant turions of 22 aquatic plant species: do they reflect functional or taxonomic traits?

Turions are vegetative, dormant overwintering organs formed in aquatic plants in response to unfavourable ecological conditions. Contents of cytokinin (CK), auxin metabolites and abscisic acid (ABA) as main growth and development regulators were compared in innately dormant autumnal turions of 22 aquatic plant species of different functional ecological or taxonomic groups with those in non-dormant winter apices in three aquatic species and with those in spring turions of four species after their overwintering. The hormones were analysed in miniature turion samples using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry. In innately dormant turions, the total contents of each of the four main CK types, biologically active forms and total CKs differed by two to three orders of magnitude across 22 species; the proportion of active CK forms was 0.18-67 %. Similarly, the content of four auxin forms was extremely variable and the IAA proportion as the active form was 0.014-99 %. The ABA content varied from almost zero to 54 µmol kg-1 dry weight and after overwintering it usually significantly decreased. Of all functional traits studied, hormone profiles depended most on the place of turion sprouting (surface vs bottom) and we suggest that this trait is crucial for turion ecophysiology. The key role of ABA in regulating turion dormancy was confirmed. However, the highly variable pattern of the ABA content in innately dormant and in overwintered turions indicates that the hormonal mechanism regulating the innate dormancy and its breaking in turions is not uniform within aquatic plants.

Meta-DHGNN: method for CRS-related cytokines analysis in CAR-T therapy based on meta-learning directed heterogeneous graph neural network.

Chimeric antigen receptor T-cell (CAR-T) immunotherapy, a novel approach for treating blood cancer, is associated with the production of cytokine release syndrome (CRS), which poses significant safety concerns for patients. Currently, there is limited knowledge regarding CRS-related cytokines and the intricate relationship between cytokines and cells. Therefore, it is imperative to explore a reliable and efficient computational method to identify cytokines associated with CRS. In this study, we propose Meta-DHGNN, a directed and heterogeneous graph neural network analysis method based on meta-learning. The proposed method integrates both directed and heterogeneous algorithms, while the meta-learning module effectively addresses the issue of limited data availability. This approach enables comprehensive analysis of the cytokine network and accurate prediction of CRS-related cytokines. Firstly, to tackle the challenge posed by small datasets, a pre-training phase is conducted using the meta-learning module. Consequently, the directed algorithm constructs an adjacency matrix that accurately captures potential relationships in a more realistic manner. Ultimately, the heterogeneous algorithm employs meta-photographs and multi-head attention mechanisms to enhance the realism and accuracy of predicting cytokine information associated with positive labels. Our experimental verification on the dataset demonstrates that Meta-DHGNN achieves favorable outcomes. Furthermore, based on the predicted results, we have explored the multifaceted formation mechanism of CRS in CAR-T therapy from various perspectives and identified several cytokines, such as IFNG (IFN-γ), IFNA1, IFNB1, IFNA13, IFNA2, IFNAR1, IFNAR2, IFNGR1 and IFNGR2 that have been relatively overlooked in previous studies but potentially play pivotal roles. The significance of Meta-DHGNN lies in its ability to analyze directed and heterogeneous networks in biology effectively while also facilitating CRS risk prediction in CAR-T therapy.

Characteristics of turion development in two aquatic carnivorous plants: Hormonal profiles, gas exchange and mineral nutrient content.

Turions are vegetative, dormant, and storage overwintering organs formed in perennial aquatic plants in response to unfavorable ecological conditions and originate by extreme condensation of apical shoot segments. The contents of cytokinins, auxins, and abscisic acid were estimated in shoot apices of summer growing, rootless aquatic carnivorous plants, Aldrovanda vesiculosa and Utricularia australis, and in developing turions at three stages and full maturity to reveal hormonal patterns responsible for turion development. The hormones were analyzed in miniature turion samples using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry. Photosynthetic measurements in young leaves also confirmed relatively high photosynthetic rates at later turion stages. The content of active cytokinin forms was almost stable in A. vesiculosa during turion development but markedly decreased in U. australis. In both species, auxin content culminated in the middle of turion development and then decreased again. The content of abscisic acid as the main inhibitory hormone was very low in growing plants in both species but rose greatly at first developmental stages and stayed very high in mature turions. The hormonal data indicate a great strength of developing turions within sink-source relationships and confirm the central role of abscisic acid in regulating the turion development.

Profiling of adenine-derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide-induced cell stress.

Cytokinins (CTKs) are a diverse collection of evolutionarily conserved adenine-derived signaling molecules classically studied as phytohormones; however, their roles and production have been less studied in mammalian systems. Skeletal muscles are sensitive to cellular cues such as inflammation and in response, alter their secretome to regulate the muscle stem cell and myofiber niche. Using cultured C2C12 muscle cells, we profiled CTK levels to understand (1) whether CTKs are part of the muscle secretome and (2) whether CTKs are responsive to cellular stress. To induce cellular stress, C2C12 myotubes were treated with lipopolysaccharides (LPS) for 24 h and then media and cell fractions were collected for ultra high-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC-(ESI+)-HRMS/MS) for metabolomics and CTK profiling. Across LPS-treated and control cells, 11 CTKs were detected in the extracellular space while 6 were detected intracellularly. We found that muscle cells are enriched in isopentenyladenine (iP) species (from free base, riboside to nucleotide forms), and that extracellular levels are increased after LPS treatment. Our study establishes that muscle cells express various forms of CTKs, and that CTK levels are responsive to LPS-induced cell stress, suggesting a role for CTKs in intra- and extracellular signaling of mammalian cells.

Peculiarities of soybean-rhizobial systems subject to different levels of water supply fol-lowing treatment with succinic acid and epibrassinolide

All around the world, one of the leading – according to area of cultivated fields – oleic crops is soybean, which has a high demand for moisture. Given the significance of this crop and negative impact of drought on its yield, integrated research of the influence of insufficient water supply on the intensity of physiological-biochemical processes in those plants is necessary for identifying and understanding the drought-tolerance mechanisms of soybean, as well as symbiotic systems created with its participation, and also for search for ways to adapt it to this stressor. Therefore, our objective was determining the specifics of formation and functioning of the symbiotic systems of soybean and Bradyrhizobium japonicum, following treatment with succinic acid (0.01 g/L) and 24-epibrassinolide (0.00001 g/L), subject to different levels of watering. Our studies revealed that pre-sowing treatment of the seeds with a solution of 24-epibrassinolide with their subsequent inoculation with B. japonicum Т21-2 resulted in the most pronounced stimulation of formation and functioning of the symbiotic systems of soybean in the optimal growing conditions. At the same time, during water shortage, the intensity of nitrogen fixation was the highest in the plants grown from seeds that had been successively treated with the acid and the inoculant. We confirmed that water deficit led to significant increase in the overall content of phytohormones of cytokinin nature in the soybean root nodules, depending on the way the seeds were treated. However, the largest pool of cytokinins was seen in the plants that had been treated with succinic acid against the background of both optimal and insufficient water supply. Treatment of the seeds with 24-epibrassinolide caused significant excess of content of zeatin riboside over the content of zeatin during the flowering stage, whereas in the stage of pods formation it led to an opposite effect – excess of zeatin over zeatin riboside. Fourteen days-long water deficit decreased the content of chlorophylls in the leaves and grain productivity of the plants of all variants of the experiment. The use of growth regulators managed to alleviate the negative impact of stress and protect the pigment complex from ruination. Treatment of the seeds with solutions of succinic acid and 24-epibrassinolide provided the growth of soybean grain productivity regardless on water-supply level. The most efficient was 24-epibrassinolide. Therefore, use of 24-epibrassinolide for pre-sowing treatment of the soybean seeds provided formation of effective symbiotic systems with high nitrogen-fixing activity and caused a number of specific changes in the pattern of accumulation of free and complex forms of cytokinins in the root nodules of those plants. At the same time, the treatment provided the highest concentration of photosynthesis pigments in the soybean leaves, and as a result produced the greatest increase in grain productivity of plants of all the variants, regardless of levels of water supply. In turn, use of succinic acid produced the highest level of nitrogen-fixing activity in the case of the lowest number of root nodules in the conditions of insufficient water supply, and also caused significant accumulation of cytokinins in the nodules, compared with other studied variants against the background of both optimal and insufficient water supply. Therefore, it did result in increase in soybean grain productivity, but this was lower than in the plants treated with 24-epibrassinolide.

Cytokinin Translocation to, and Biosynthesis and Metabolism within, Cereal and Legume Seeds: Looking Back to Inform the Future.

Early in the history of cytokinins, it was clear that Zea mays seeds contained not just trans-zeatin, but its nucleosides and nucleotides. Subsequently, both pods and seeds of legumes and cereal grains have been shown to contain a complex of cytokinin forms. Relative to the very high quantities of cytokinin detected in developing seeds, only a limited amount appears to have been translocated from the parent plant. Translocation experiments, and the detection of high levels of endogenous cytokinin in the maternal seed coat tissues of legumes, indicates that cytokinin does not readily cross the maternal/filial boundary, indicating that the filial tissues are autonomous for cytokinin biosynthesis. Within the seed, trans-zeatin plays a key role in sink establishment and it may also contribute to sink strength. The roles, if any, of the other biologically active forms of cytokinin (cis-zeatin, dihydrozeatin and isopentenyladenine) remain to be elucidated. The recent identification of genes coding for the enzyme that leads to the biosynthesis of trans-zeatin in rice (OsCYP735A3 and 4), and the identification of a gene coding for an enzyme (CPN1) that converts trans-zeatin riboside to trans-zeatin in the apoplast, further cements the key role played by trans-zeatin in plants.

Are Histidine Kinases of Arbuscular Mycorrhizal Fungi Involved in the Response to Ethylene and Cytokinins?

Signals are exchanged at all stages of the arbuscular mycorrhizal (AM) symbiosis between fungi and their host plants. Root-exuded strigolactones are well-known early symbiotic cues, but the role of other phytohormones as interkingdom signals has seldom been investigated. Here we focus on ethylene and cytokinins, for which candidate receptors have been identified in the genome of the AM fungus Rhizophagus irregularis. Ethylene is known from the literature to affect asymbiotic development of AM fungi, and in the present study, we found that three cytokinin forms could stimulate spore germination in R. irregularis. Heterologous complementation of a Saccharomyces cerevisiae mutant strain with the candidate ethylene receptor RiHHK6 suggested that this protein can sense and transduce an ethylene signal. Accordingly, its N-terminal domain expressed in Pichia pastoris displayed saturable binding to radiolabeled ethylene. Thus, RiHHK6 displays the expected characteristics of an ethylene receptor. In contrast, the candidate cytokinin receptor RiHHK7 did not complement the S. cerevisiae mutant strain or Medicago truncatula cytokinin receptor mutants and seemed unable to bind cytokinins, suggesting that another receptor is involved in the perception of these phytohormones. Taken together, our results support the hypothesis that AM fungi respond to a range of phytohormones and that these compounds bear multiple functions in the rhizosphere beyond their known roles as internal plant developmental regulators. Our analysis of two phytohormone receptor candidates also sheds new light on the possible perception mechanisms in AM fungi. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Temporal and spatial transcriptional regulation of phytohormone metabolism during seed development in barley (Hordeum vulgare L.).

Plant hormones play important roles in seed development; however, transcriptional regulation of their metabolism and levels of the respective bioactive forms during barley seed development is poorly understood. To this end, this study performed a comprehensive analysis of changes in the expression patterns phytohormone metabolism genes and levels of the respective bioactive forms in the embryo and endosperm tissues. Our study showed the presence of elevated levels of abscisic acid (ABA), bioactive forms of gibberellins (GAs), jasmonate (JA) and cytokinins (CKs), auxin and salicylic acid (SA) in the endosperm and embryo tissues at early stage of seed filling (SF). The levels of all hormones in both tissues, except that of ABA, decreased to low levels during SF. In contrast, embryonic ABA level increased during SF and peaked at physiological maturity (PM) while the endospermic ABA was maintained at a similar level observed during SF. Although its level decreased high amount of ABA was still present in the embryo during post-PM. We detected low levels of ABA in the endosperm and all the other hormones in both tissues during post-PM phase except the relatively higher levels of jasmonoyl-isoleucine and SA detected at late stage of post-PM. Our data also showed that spatiotemporal changes in the levels of plant hormones during barley seed development are mediated by the expression of specific genes involved in their respective metabolic pathways. These results indicate that seed development in barley is mediated by spatiotemporal modulation in the metabolism and levels of plant hormones.

Phytohormones enhance heavy metal responses in Euglena gracilis: Evidence from uptake of Ni, Pb and Cd and linkages to hormonomic and metabolomic dynamics

Over the last decade, significant effort has been made to understand phytohormonal functions (e.g., cytokinins (CKs) and abscisic acid (ABA)) in metal stress responses of higher plants and algae. Despite the potential for these phytohormones to improve industrial remediation by Euglena gracilis (Euglenophyceae), no such roles have been elucidated for this highly adaptive species and its response to heavy metals. This study demonstrates that toxic metals (nickel, lead, cadmium) modify hormonal activity profiles (i.e., CK forms and their concentrations) in E. gracilis. Furthermore, exogenous ABA or CK (tZ) enabled higher metal uptake efficiency (i.e., 9.35% in lead and 9.2% in cadmium uptake with CK) and alleviated metal toxicity through the regulation of endogenous CKs (i.e., total CK, isoprenoid CK) and gibberellin (GAs, GA1 and GA3) levels. These responses suggest that E. gracilis regulates multiple phytohormone signals during metal stress acclimation. A deeper approach, using untargeted metabolomic analyses, gave more detailed insight into phytohormone-controlled pathways and associated modified metabolites, which were frequently related to metal accumulation and the physiological acclimation to metal presence. Significant changes in the levels of cellular metabolites, especially those involved in acclimation to metal stress, were under the influence of phytohormones in algal cells. When grown under metal stress conditions, the presence of exogenous ABA or CKs, caused changes in cellular metabolites which included those from: lipid pathways, riboflavin metabolism, the biosynthesis of cofactors/vitamins, and carbohydrate metabolism. Also, bioactive secondary metabolites (e.g., terpenoids, alkaloids, flavonoids, carotenoids) were modified in algal cells treated with phytohormones. Thus, the study gives a detailed view on the regulatory functions of ABA and CKs in algal metal bioremediation strategies, which are attributed to enhanced metal uptake and in the fine-tuning of plant hormone levels during metal stress response. The results can guide efforts to develop efficient, low-cost and environmentally friendly methods for bioremediation.

Cytokinins and auxins in organs of aquatic carnivorous plants: what do they reflect?

Aquatic carnivorous plants have typical rootless linear shoots bearing traps and exhibit steep physiological polarity with rapid apical growth. The aim was to analyse auxin and cytokinin metabolites in traps, leaves/shoots and shoot apices in several species of genera Aldrovanda and Utricularia to elucidate how the hormonal profiles reflect the specific organ functions and polarity. The main auxin and cytokinin metabolites were analysed in miniature samples (>2 mg dry weight) of different organs of Aldrovanda vesiculosa and six Utricularia species using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry. Total contents of biologically active forms (free bases, ribosides) of all four main endogenously occurring cytokinin types were consistently higher in traps than in leaves in four Utricularia species with monomorphic shoots and/or higher than in shoots in two Utricularia species with dimorphic shoots. In Aldrovanda traps, the total content of different cytokinin forms was similar to or lower than that in shoots. In U. australis leaves, feeding on prey increased all cytokinin forms, while no consistent differences occurred in Aldrovanda. In four aquatic Utricularia species with monomorphic shoots, the content of four auxin forms was usually higher in traps than in leaves. Zero IAA content was determined in U. australis leaves from a meso-eutrophic site or when prey-fed. Different cytokinin and auxin profiles estimated in traps and leaves/shoots of aquatic carnivorous plants indicate an association with different dominant functions of these organs: nutrient uptake by traps versus photosynthetic function of traps. Interplay of cytokinins and auxins regulates apical dominance in these plants possessing strong polarity.

Spatiotemporal NF-κB dynamics encodes the position, amplitude, and duration of local immune inputs.

Infected cells communicate through secreted signaling molecules like cytokines, which carry information about pathogens. How differences in cytokine secretion affect inflammatory signaling over space and how responding cells decode information from propagating cytokines are not understood. By computationally and experimentally studying NF-κB dynamics in cocultures of signal-sending cells (macrophages) and signal-receiving cells (fibroblasts), we find that cytokine signals are transmitted by wave-like propagation of NF-κB activity and create well-defined activation zones in responding cells. NF-κB dynamics in responding cells can simultaneously encode information about cytokine dose, duration, and distance to the cytokine source. Spatially resolved transcriptional analysis reveals that responding cells transmit local cytokine information to distance-specific proinflammatory gene expression patterns, creating “gene expression zones.” Despite single-cell variability, the size and duration of the signaling zone are tightly controlled by the macrophage secretion profile. Our results highlight how macrophages tune cytokine secretion to control signal transmission distance and how inflammatory signaling interprets these signals in space and time.

Profiling of phytohormones in apple fruit and buds regarding their role as potential regulators of flower bud formation.

Apple (Malus×domestica Borkh.) cropping behavior, if not regulated, is often manifested by high yields of small-sized fruit in so called ON-years, which are usually followed by strongly reduced crop loads in OFF-years. Such cropping pattern is defined as biennial bearing and causes significant losses in apple production. The growth of apple fruit overlaps with the formation of flower buds, which remain dormant until the following spring. Earlier works proposed that some fruit-derived mobile compounds, as e.g., phytohormones, could suppress flower bud formation that thereby leads to biennial bearing. We addressed this hypothesis by analyzing 39 phytohormones in apple seeds, fruit flesh and by measuring phytohormone export from the fruits of the biennial bearing cultivar 'Fuji' and of the regular bearing cultivar 'Gala'. Moreover, we analyzed the same compounds in bourse buds from fruiting (ON-trees) and non-fruiting (OFF-trees) spurs of both apple cultivars over the period of flower bud formation. Our results showed that apple fruit exported at least 14 phytohormones including indole-3-acetic acid and gibberellin A3; however, their influence on flower bud formation was inconclusive. A gibberellin-like compound, which was detected exclusively in bourse buds, was significantly more abundant in bourse buds from ON-trees compared with OFF-trees. Cultivar differences were marked by the accumulation of trans-zeatin-O-glucoside in bourse buds of 'Gala' ON-trees, whereas the levels of this compound in 'Gala' OFF were significantly lower and comparable to those in 'Fuji' ON- and OFF-trees. Particular phytohormones including five cytokinin forms as well as abscisic acid and its degradation products had higher levels in bourse buds from OFF-trees compared with ON-trees and were therefore proposed as potential promotors of flower bud initiation. The work discusses regulatory roles of phytohormones in flower bud formation in apple based on the novel and to date most comprehensive phytohormone profiles of apple fruit and buds.

Phytohormones in galls on eucalypt trees and in the gall‐forming wasp Leptocybe invasa (Hymenoptera: Eulophidae)

Abstract The role played by phytohormones in insect galls and the mechanisms underlying plant manipulation by gall makers are still unclear. Galls made by Leptocybe invasa on two Eucalyptus species were sampled, the correlations between the levels of nine phytohormones in galls and gall size were analysed, and phytohormone levels in the interior and exterior tissues of galls and L. invasa were determined. During two larval stages, the levels of three forms of cytokinins (CKs) in the galls of the two plant species all showed significantly positive correlations with gall size. Brassinosteroids (BRs) and dihydrozeatin riboside (DHZR) occurred at significantly higher levels in the interior tissues of the galls compared to in the exterior tissues of galls. Isopentenyl adenosine (IPA) was significantly higher in L. invasa larvae themselves, compared to levels in the interior and exterior tissues of galls. However, the levels of DHZR and BR were significantly lower or similar in the L. invasa pupae compared to that in the interior and exterior tissues of the galls. These findings indicate that phytohormones, especially CKs, act as key regulators of L. invasa galls, and that the larvae of L. invasa may themselves be a source of CKs.

Morphogenesis, pigment content, phytohormones and yield of tomatoes under the action of gibberellin and tebuconazole

One of the main tasks of contemporary plant physiology is regulation of growth and development of cultivated plants in order to optimize the productive process. The scientific community focuses its attention on the use of natural activators and growth inhibitors. We investigated the effect of foliar treatment with 0.005% solution of gibberellic acid and 0.025% solution of the anti-gibberellic preparation tebuconazole on morphogenesis, leaf mesostructure, content of photosynthetic pigments, balance of endogenous phytobrybrides and lymphocyte B and productivity of tomatoes. The vegetation experiment was carried out in the conditions of soil-sand culture in vessels with a 10-liter volume. The treatment was carried out in the budding phase. Morphometric parameters were measured every 10 days. The mesostructure of the middle tier leaves was studied in the fruit formation phase, and the chlorophyll content was determined in the raw material by spectrophotometric method. Analytical determination of endogenous phytohormones – indolyl-3-acetic (IAA), gibberellic acid and abscisic (ABA) acids and cytokinins – zeatin (Z), zeatin-O-glucoside (ZG), zeatinribozide (Znila) and isopentenyladenosine (iPA) was performed by high performance liquid chromatography – mass spectrometry (HPLC-MS). With gibberellic acid treatment plant height increased significantly, while with tebuconazole it decreased. Gibberellic acid increased the number of leaves per plant, and tebuconazole did not change it. The preparations increased the number of leaf blades per leaf, the total number of leaf blades per plant, the weight of the raw material of leaves, the area of leaf blades and the area of the leaves at the end of the study period. The dry matter weight of stems and roots under the action of gibberellic acid increased, and during the treatment of tebuconazole decreased. Gibberellic acid increased the dry matter of the whole plant, and tebuconazole did not change it. Under the action of tebuconazole the content of chlorophyll in the leaves increased, while under the action of gibberellic acid it decreased. Both regulators increased the volume of columnar parenchyma cells. Gibberellic acid increased the size of spongy parenchyma cells, while tebuconazole did not change them. It is revealed that the action of exogenous gibberellic acid in stems and leaves increased the content of endogenous IAA and gibberellic acid, and tebuconazole decreased their content. The ABA content in stems and leaves increased with tebuconazole treatments and decreased with exogenous gibberellic acid. The total cytokinin content in the leaves was higher than in the stems in both the control and the experiment samples. Growth regulators induced an increase in the cytokinin pool in leaves and a decrease in stems. Gibberellic acid increased the content of all five forms of cytokinins in the leaves, and tebuconazole increased only two isoforms. In the stems under the action of both growth regulators the content of Z decreased and iP increased. The content of ZR and iPA in stems increased after the application of the retardant and decreased under the action of growth stimulant. The ZG content exceeded the control after gibberellic acid treatment and was in trace concentrations under the action of tebuconazole. Growth regulators optimized the productivity of tomato plants: under the action of gibberellic acid there was a considerable increase in the number of fruits per plant, and after the use of tebuconazole the average weight of one fruit significantly increased. The obtained results demonstrated that anatomical-morphological and structural-functional rearrangements in tomato plants under the action of exogenous gibberellic acid and tebuconazole occurred against the background of changes in the balance and distribution of endogenous hormones. Increased photosynthetic activity, stimulation of growth processes of some plant organs and inhibition of others increased the biological crop capacity.

Secondary Active Transporters Decrease Cytokinin Flow into Barley Shoots and Inhibit their Growth under Phosphate Deficit

Secondary active transport is one of the most important mechanisms controlling cytokinin distribution between shoots and roots which enables the adaptive growth reaction of barley. Concentrations (ng/g of fresh mass) and contents (ng/organ) of different forms of cytokinins were determined in shoots and roots of barley plants by immunoassay. It was shown that under phosphate deficit the maintenance of root growth, inhibition of root branching and decline in shoot mass of barley plants (Hordeum vulgare 'Prairie') was due to decline in shoot cytokinins and their accumulation in the roots. Protonophore carbonyl cyanide-m-chlorophenylhydrazone as an inhibitor of secondary active transport leveled off hormonal reaction to phosphate deficit: it prevented accumulation of cytokinins in roots, increased their content in shoots and changed the percentage of zeatin and its derivatives in total cytokinin content in roots and shoots. A data analysis showed that in roots, protonophore did not change significantly the content of ribosides and nucleotides of zeatin, while uptake of free zeatin by cells was significantly affected by the protonophore and its content decreased dramatically. The role of secondary active trans-membrane transfer is discussed in the context of cytokinins transport from roots to shoots under phosphate deficit

Developmental and hormonal regulation of Arabidopsis thaliana ornithine-delta-aminotransferase.

Ornithine aminotransferase (OAT) catalyzes transfer of the delta-amino group from L-ornithine to oxo-glutarate. In plants, this reaction biochemically connects urea cycle, proline cycle, and polyamine biosynthesis pathway. OAT activity is shown to be associated with biotic and abiotic stress responses and nitrogen metabolism, but its physiological role is still unclear. In our study, we decided to investigate transcriptional regulation of the OAT gene in Arabidopsis thaliana under normal conditions and in response to various growth regulators. In the present work, the reporter gene construct containing the Escherichia coli β-glucuronidase gene (gus) under control of the A. thaliana OAT gene promoter was introduced into the genome of A. thaliana ecotype Columbia plants using the floral dip method; GUS activity was assayed in different experimental conditions including hormone treatment, low and high nitrogen and salinity. The GUS activity was analyzed histochemically. Plants were incubated with staining solution containing X-Gluc. We show that under standard growth conditions, the promoter is active during germination and in developing floral organs. OAT promoter activity specifically activates in response to different forms of auxin (IAA, NAA, and 2,4D), cytokinin (6- BAP), ethylene precursor (ACC), high nitrogen and salinity. Analysis of the OAT expression by qRT-PCR confirmed the pattern observed using the GUS reporter system. The OAT gene showed a significantly elevated expression in fourday- old seedlings and in plant roots in response to auxins and cytokinins. The analysis of the OAT promoter structure reveals cis-acting regulatory DNA elements associated with auxin regulation and abiotic stresses. The results of the study indicate that the OAT gene is involved in developmental processes and is regulated by auxin and cytokinins.

Identification of Potential Cytokinin Responsive Key Genes in Rice Treated With Trans-Zeatin Through Systems Biology Approach.

Rice is an important staple food grain consumed by most of the population around the world. With climate and environmental changes, rice has undergone a tremendous stress state which has impacted crop production and productivity. Plant growth hormones are essential component that controls the overall outcome of the growth and development of the plant. Cytokinin is a hormone that plays an important role in plant immunity and defense systems. Trans-zeatin is an active form of cytokinin that can affect plant growth which is mediated by a multi-step two-component phosphorelay system that has different roles in various developmental stages. Systems biology is an approach for pathway analysis to trans-zeatin treated rice that could provide a deep understanding of different molecules associated with them. In this study, we have used a weighted gene co-expression network analysis method to identify the functional modules and hub genes involved in the cytokinin pathway. We have identified nine functional modules comprising of different hub genes which contribute to the cytokinin signaling route. The biological significance of these identified hub genes has been tested by applying well-proven statistical techniques to establish the association with the experimentally validated QTLs and annotated by the DAVID server. The establishment of key genes in different pathways has been confirmed. These results will be useful to design new stress-resistant cultivars which can provide sustainable yield in stress-specific conditions.

Systemic inflammation alters the neuroinflammatory response: a prospective clinical trial in traumatic brain injury

BackgroundNeuroinflammation following traumatic brain injury (TBI) has been shown to be associated with secondary injury development; however, how systemic inflammatory mediators affect this is not fully understood. The aim of this study was to see how systemic inflammation affects markers of neuroinflammation, if this inflammatory response had a temporal correlation between compartments and how different compartments differ in cytokine composition.MethodsTBI patients recruited to a previous randomised controlled trial studying the effects of the drug anakinra (Kineret®), a human recombinant interleukin-1 receptor antagonist (rhIL1ra), were used (n = 10 treatment arm, n = 10 control arm). Cytokine concentrations were measured in arterial and jugular venous samples twice a day, as well as in microdialysis-extracted brain extracellular fluid (ECF) following pooling every 6 h. C-reactive protein level (CRP), white blood cell count (WBC), temperature and confirmed systemic clinical infection were used as systemic markers of inflammation. Principal component analyses, linear mixed-effect models, cross-correlations and multiple factor analyses were used.ResultsJugular and arterial blood held similar cytokine information content, but brain-ECF was markedly different. No clear arterial to jugular gradient could be seen. No substantial delayed temporal associations between blood and brain compartments were detected. The development of a systemic clinical infection resulted in a significant decrease of IL1-ra, G-CSF, PDGF-ABBB, MIP-1b and RANTES (p < 0.05, respectively) in brain-ECF, even if adjusting for injury severity and demographic factors, while an increase in several cytokines could be seen in arterial blood.ConclusionsSystemic inflammation, and infection in particular, alters cytokine levels with different patterns seen in brain and in blood. Cerebral inflammatory monitoring provides independent information from arterial and jugular samples, which both demonstrate similar information content. These findings could present potential new treatment options in severe TBI patients, but novel prospective trials are warranted to confirm these associations.Graphical abstract