Soil biological activity after a sixty-year fertilization practice in a wheat-maize crop rotation.

This study aimed to survey the long-term effects of fertilization practices on the functional diversity of the soil microbiota. A 60-year fertilization experiment with mineral fertilizers, farmyard manure and combined treatments was sampled in two consecutive years in maize (Zea mays L.) and wheat (Triticum aestivum L.). Soil chemical properties, plant growth and physiological parameters were measured. The MicroRespTM method was applied to assess the community level physiological profiles (CLPPs) of the rhizosphere soil, and the arbuscular mycorrhizal fungal (AMF) colonization of the roots was determined. Samples were taken in the early vegetative stages, at flowering, and at harvest in both years. The measured parameters were analysed using multifactorial ANOVA to determine treatment effects, crop-dependent differences, and seasonality. PCA analysis was performed on the data matrix to reveal more complex correspondences, and Pearson's product-moment correlation was used to confirm relationships between some of the measured soil and plant parameters. Fertilization treatments caused long-term changes in some biological parameters such as: MicroRespTM parameters, citrate utilization, total substrate-induced respiration value, and the ratio of utilization of amino acids and sugars. The rate of AMF colonization responded mainly to the plant nutrition status and the plant requirements, suggesting a plant-mediated effect in the case of mycorrhiza. Mineral nitrogen fertilization and soil acidification were found to be the main factors affecting the catabolic activity of soil microbiota, while AMF colonization responded to the balance of plant nutrition.

Comprehensive analysis of different tumor cell-line produced soluble mediators on the differentiation and functional properties of monocyte-derived dendritic cells.

Developing dendritic cells (DCs) from monocytes is a sensitively regulated process. One possible way for cancers to avoid immune recognition and antitumor response is the modulation of DC differentiation. Although several studies are available on the examination of tumor-associated macrophages, a comprehensive analysis focusing on the effects of tumor-formed DCs is not known to date. We provide a comparative analysis of the tumor-edited-monocyte derived DCs differentiated in the presence of adenocarcinomas (MDA, HT29, HeLa)- and primary (WM278, WM983A) or metastatic (WM1617, WM983B) melanomas. The immunomodulatory effect of tumors is mediated at least partly by secreted mediators. We investigated the impact of tumor cell-derived conditioned media on the differentiation of DCs from CD14+ monocytes, sequentially determining the phenotype, cytokine production, phagocytic, and the T cell polarizing capacity of moDCs. We completed our observations by analyzing our data with bioinformatic tools to provide objective correlations between phenotypical and functional properties of different tumor-educated moDCs. The correlation analysis revealed significant differences in the characteristics of adenocarcinomas- or melanomas-edited moDCs. We highlight the functional differences in the properties of moDCs differentiated in the presence of various cancer cell lines. We offer new information and options for the in vitro differentiation protocols of various tumor-conditioned moDCs. Our results confirm that various immunomodulatory properties of different tumor cell lines result in multiple manipulations of DC differentiation.

Electrical Capacitance versus Minirhizotron Technique: A Study of Root Dynamics in Wheat-Pea Intercrops.

This study evaluated the concurrent application and the results of the root electrical capacitance (CR) and minirhizotron (MR) methods in the same plant populations. The container experiment involved three winter wheat cultivars, grown as sole crops or intercropped with winter pea under well-watered or drought-stressed conditions. The wheat root activity (characterized by CR) and the MR-based root length (RL) and root surface area (RSA) were monitored during the vegetation period, the flag leaf chlorophyll content was measured at flowering, and the wheat shoot dry mass (SDM) and grain yield (GY) were determined at maturity. CR, RL and RSA exhibited similar seasonal patterns with peaks around the flowering. The presence of pea reduced the maximum CR, RL and RSA. Drought significantly decreased CR, but increased the MR-based root size. Both intercropping and drought reduced wheat chlorophyll content, SDM and GY. The relative decrease caused by pea or drought in the maximum CR was proportional to the rate of change in SDM or GY. Significant linear correlations (R2: 0.77-0.97) were found between CR and RSA, with significantly smaller specific root capacitance (per unit RSA) for the drought-stress treatments. CR measurements tend to predict root function and the accompanying effect on above-ground production and grain yield. The parallel application of the two in situ methods improves the evaluation of root dynamics and plant responses.

MSC-like cells increase ability of monocyte-derived dendritic cells to polarize IL-17-/IL-10-producing T cells via CTLA-4.

Mesenchymal stromal cell-like (MSCl) cells generated from human embryonic stem cells are considered to be an eligible cell line to model the immunomodulatory behavior of mesenchymal stromal cells (MSCs) in vitro. Dendritic cells (DCs) are essential players in the maintenance and restoration of the sensitive balance between tolerance and immunity. Here, the effects of MSCl cells on the in vitro differentiation of human monocytes into DCs were investigated. MSCl cells promote the differentiation of CTLA-4 expressing DCs via the production of all-trans retinoic acid (ATRA) functioning as a ligand of RARα, a key nuclear receptor in DC development. These semi-matured DCs exhibit an ability to activate allogeneic, naive T cells and polarize them into IL-10 + IL-17 + double-positive T helper cells in a CTLA-4-dependent manner. Mapping the molecular mechanisms of MSC-mediated indirect modulation of DC differentiation may help to expand MSCs' clinical application in cell-free therapies.

Differences in the sensitivity of classically and alternatively activated macrophages to TAK1 inhibitor-induced necroptosis.

Controlling the balance of pro-inflammatory M1 versus anti-inflammatory M2 macrophages may have paramount therapeutic benefit in cardiovascular diseases, infections, cancer and chronic inflammation. The targeted depletion of different macrophage populations provides a therapeutic option to regulate macrophage-mediated functions. Macrophages are highly sensitive to necroptosis, a newly described regulated cell death mediated by receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3 and mixed lineage kinase domain like pseudokinase. Antagonists of inhibitors of apoptosis proteins (SMAC mimetics) block RIPK1 ubiquitination, while TGF-activated kinase 1 (TAK1) inhibitors prevent the phosphorylation of RIPK1, resulting in increased necroptosis. We compared the sensitivity of monocyte-derived human M1 and M2 cells to various apoptotic and necroptotic signals. The two cell types were equally sensitive to all investigated stimuli, but TAK1 inhibitor induced more intense necroptosis in M2 cells. Consequently, the treatment of co-cultured M1 and M2 cells with TAK1 inhibitor shifted the balance of the two populations toward M1 dominance. Blockage of either Aurora Kinase A or glycogen synthase kinase 3ß, two newly described necroptosis inhibitors, increased the sensitivity of M1 cells to TAK1-inhibitor-induced cell death. Finally, we demonstrated that in vitro differentiated tumor-associated macrophages (TAM-like cells) were as highly sensitive to TAK1 inhibitor-induced necroptosis as M2 cells. Our results indicate that at least two different necroptotic pathways operate in macrophages and the targeted elimination of different macrophage populations by TAK1 inhibitor or SMAC mimetic may provide a therapeutic option to regulate the balance of inflammatory/anti-inflammatory macrophage functions.

Density and Diversity of Microbial Symbionts under Organic and Conventional Agricultural Management.

The influence of organic and conventional farming and agroecology on the diversity and functioning of indigenous soil microbial communities was examined using a multifactorial analysis based on an extended minimum data set of classical status and functional tests. Main soil physicochemical properties and selected microbiological indicators, the quantity of heterotrophic or aerobic spore-forming bacteria, basal and substrate-induced respiration, catabolic activity with MicroResp™, and fluorescein diacetate enzyme activity were characterized. A pot experiment applying the most probable number method was designed with soil dilution series using Pisum sativum L. and Triticum spelta L. to assess the symbiotic infectivity and genetic diversity of key indicator groups of the plant microbiome, e.g. nitrogen-fixing bacteria (rhizobia) and arbuscular mycorrhizal fungi. Soil pH, humus content, CFU, enzyme activity, and soil respiration were significantly higher in organic soils. The activity of soil microorganisms was mainly related to clay, humus, calcium, and magnesium parameters. A redundancy analysis test of catabolic activities showed that samples were grouped according to different substrate utilization patterns and land uses were also clearly separated from each other. Farming practice influenced the abundance and diversity of microbial populations. Dark septate endophytic fungi were only found in conventional soils. In addition to confirming soil health improvements by organic management, our results highlight the importance of a complex evaluation including both classical status and functional parameters of soil microbiota, which may more reliably indicate a shift in the quality status of soils.

Symbiotic Effectivity of Dual and Tripartite Associations on Soybean (Glycine max L. Merr.) Cultivars Inoculated With Bradyrhizobium japonicum and AM Fungi.

Soybean (Glycine max L. Merr.) is regarded worldwide as indisputably one of the most important crops for human food and animal feed. The presence of symbiotic bacteria and fungi is essential for soybean breeding, especially in low-input agricultural systems. Research on the cooperation between different microbial symbionts is a key to understanding how the health and productivity of the plant is supported. The symbiotic effectivity of dual and tripartite symbiotic agents was investigated in two pot experiments on different soybean cultivars with special regard to compatibility. In the Selection experiment, two out of sixteen soybean cultivars (Aliz, Emese) were chosen on the basis of their drought tolerance and used in all the other investigations. In the Compatibility experiment, the compatible coupling of symbiotic partners was selected based on the efficiency of single and co-inoculation with two Bradyrhizobium japonicum strains and two commercial arbuscular mycorrhizal fungal (AMF) products. Significant differences were found in the infectivity and effectivity of the microsymbionts. The rhizobial and AMF inoculation generally improved plant production, photosynthetic efficiency and root activity, but this effect depended on the type of symbiotic assotiation. Despite the low infectivity of AMF, inocula containing fungi were more beneficial than those containing only rhizobia. In the Drought Stress (DS) experiment, co-inoculated and control plants were grown in chernozem soil originating from organic farms. Emese was more resistant to drought stress than Aliz and produced a bigger root system. Under DS, the growth parameters of both microbially inoculated cultivars were better than that of control, proving that even drought tolerant genotypes can strengthen their endurance due to inoculation with AMF and nitrogen fixing bacteria. Root electrical capacitance (CR) showed a highly significant linear correlation with root and shoot dry mass and leaf area. The same root biomass was associated with higher CR in inoculated hosts. As CR method detects the absorptive surface increasing due to inoculation, it may be used to check the efficiency of the microbial treatment.

Application of Electrical Capacitance Method for Prediction of Plant Root Mass and Activity in Field-Grown Crops.

The root electrical capacitance (C R ) method is suitable for assessing root growth and activity, but soil water content (SWC) strongly influences the measurement results. This study aimed to adapt the method for field monitoring by evaluating the effect of SWC on root capacitance to ensure the comparability of C R detected at different SWC. First a pot experiment was conducted with maize and soybean to establish C R -SWC functions for the field soil. Ontogenetic changes in root activity were monitored under field conditions by simultaneously measuring C R and SWC around the roots. The C R values were normalized using SWC data and experimental C R -SWC functions to obtain C R*, the comparable indicator of root activity. The effect of arbuscular mycorrhizal fungi (AMF) inoculation on the C R* and biomass of field-grown soybean was investigated. The pot trial showed an exponential increase in C R with SWC. C R -SWC functions proved to be species-specific. C R showed strong correlation with root dry mass (R2 = 0.83-0.87). The root activity (C R*) of field-grown crops increased until flowering, then decreased during maturity. This was consistent with data obtained with other methods. AMF inoculation of soybean resulted in significantly higher C R* during the late vegetative and early flowering stages, when destructive sampling concurrently showed higher shoot biomass. The results demonstrated that the root capacitance method could be useful for time course studies on root activity under field conditions, and for comparing single-time capacitance data collected in areas with heterogeneous soil water status.

Estimation of AM fungal colonization - Comparability and reliability of classical methods.

The characterization of mycorrhizal status in hosts can be a good indicator of symbiotic associations in inoculation experiments or in ecological research. The most common microscopic-based observation methods, such as (i) the gridline intersect method, (ii) the magnified intersections method and (iii) the five-class system of Trouvelot were tested to find the most simple, easily executable, effective and objective ones and their appropriate parameters for characterization of mycorrhizal status. In a pot experiment, white clover (Trifolium repens L.) host plant was inoculated with 6 (BEG144; syn. Rhizophagus intradices) in pumice substrate to monitor the AMF colonization properties during host growth. Eleven (seven classical and four new) colonization parameters were estimated by three researchers in twelve sampling times during plant growth. Variations among methods, observers, parallels, or individual plants were determined and analysed to select the most appropriate parameters and sampling times for monitoring. The comparability of the parameters of the three methods was also tested. As a result of the experiment classical parameters were selected for hyphal colonization: colonization frequency in the first stage or colonization density in the later period, and arbuscular richness of roots. A new parameter was recommended to determine vesicule and spore content of colonized roots at later stages of symbiosis.