9 research outputs found

    Tissue Printing and Dual Excitation Flow Cytometry for Oxidative Stress—New Tools for Reactive Oxygen Species Research in Seed Biology

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    The intracellular homeostasis of reactive oxygen species (ROS) and especially of superoxide anion and hydrogen peroxide participate in signaling cascades which dictate developmental processes and reactions to stresses. ROS are also biological molecules that play important roles in seed dormancy and germination. Because of their rapid reactivity, short half-life and low concentration, ROS are difficult to measure directly with high accuracy and precision. In presented work tissue printing method with image analysis and dual excitation flow cytometry (FCM) were developed for rapid detection and localization of O2•− and H2O2 in different part of seed. Tissue printing and FCM detection of ROS showed that germination of wild oat seeds was associated with the accumulation of O2•− and H2O2 in embryo (coleorhiza, radicle and scutellum), aleurone layer and coat. To verify if printing and FCM signals were specified, the detection of O2•− and H2O2 in seeds incubated in presence of O2•− generation inhibitor (DPI) or H2O2 scavenger (CAT) were examined. All results were a high level of agreement among the level of ROS derived from presented procedures with the ones created from spectrophotometric measured data. In view of the data obtained, tissue printing with image analysis and FCM are recommended as a simple and fast methods, which could help researchers to detection and level determination of ROS in the external and inner parts of the seeds

    Nanopriming of Barley Seeds—A Shotgun Approach to Improve Germination under Salt Stress Conditions by Regulating of Reactive Oxygen Species

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    Abiotic stresses are the most important environmental factors affecting seed germination, and negatively affect crop production worldwide. Water availability is essential for proper seed imbibition and germination. The mechanism by which seeds can germinate in areas with high soil salinity is, however, still unclear. The present study aims to investigate the protective roles of AgNPs in alleviating stress symptoms caused by salinity exposure in barley seeds. For this purpose, different treatment combinations of seed priming with PVP-AgNPs in salinity stress conditions were used. Salt stress (150 and 200 mM) was found to reduce seed germination by 100% when compared to the control. Under NaCl concentrations, seed priming with PVP-AgNPs (40 mg L−1) only for 2 h, reduced salinity effects. Salinity resulted in increased reactive oxygen species (ROS) generation compared to the control. However, increased antioxidants in the NPs treatments, such as SOD, CAT, GR, GPX (expression at both genes, such as HvSOD, HvCAT, HvGR or HvGPX, and protein levels) and glutathione content, scavenged these ROS. Considering all of the parameters under study, priming alleviated salt stress. To summarize, seed priming with AgNPs has the potential to alleviate salinity stress via reduced ROS generation and activation of the antioxidant enzymatic system during germination

    Universal mtDNA fragment for Cervidae barcoding species identification using phylogeny and preliminary analysis of machine learning approach

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    Abstract The aim of the study was to use total DNA obtained from bone material to identify species of free-living animals based on the analysis of mtDNA fragments by molecular methods using accurate bioinformatics tools Bayesian approach and the machine learning approach. In our research, we present a case study of successful species identification based on degraded samples of bone, with the use of short mtDNA fragments. For better barcoding, we used molecular and bioinformatics methods. We obtained a partial sequence of the mitochondrial cytochrome b (Cytb) gene for Capreolus capreolus, Dama dama, and Cervus elaphus, that can be used for species affiliation. The new sequences have been deposited in GenBank, enriching the existing Cervidae mtDNA base. We have also analysed the effect of barcodes on species identification from the perspective of the machine learning approach. Machine learning approaches of BLOG and WEKA were compared with distance-based (TaxonDNA) and tree-based (NJ tree) methods based on the discrimination accuracy of the single barcodes. The results indicated that BLOG and WEKAs SMO classifier and NJ tree performed better than TaxonDNA in discriminating Cervidae species, with BLOG and WEKAs SMO classifier performing the best

    IoT in Water Quality Monitoring—Are We Really Here?

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    The Internet of Things (IoT) has become widespread. Mainly used in industry, it already penetrates into every sphere of private life. It is often associated with complex sensors and very complicated technology. IoT in life sciences has gained a lot of importance because it allows one to minimize the costs associated with field research, expeditions, and the transport of the many sensors necessary for physical and chemical measurements. In the literature, we can find many sensational ideas regarding the use of remote collection of environmental research. However, can we fully say that IoT is well established in the natural sciences

    Post-Effort Changes in Autophagy- and Inflammation-Related Gene Expression in White Blood Cells of Healthy Young Men

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    Acute, strenuous physical exertion requiring high levels of energy production induces the production of reactive oxygen species and metabolic disturbances that can damage the mitochondria. Thus, selective autophagic elimination of defective mitochondria may improve resistance to oxidative stress and potentially to inflammation. The main goal of this study was to evaluate the impacts of intense effort on changes in the expression of select genes related to post-effort inflammation and autophagy. Thirty-five men aged 16–21 years were recruited to the study. The impacts of both aerobic (endurance) and anaerobic (speed) efforts on selected genes encoding chemokines (CXCL5, 8–12) were analyzed. Significant increases in the expression of all studied genes excluding CXCL12 were observed. Moreover, both types of effort induced an increase in the expression of genes encoding IL-2, -4, -6, -10, IFN-γ and TNF-α, excluding IL-17A. Generally, these efforts caused a significant increase in the relative expression of apoptosis- (BCL2 and BAX) and autophagy- (BNIP3, BECN1, MAP1LC3B, ATG5, ATG7, ATG12, ATG16L1 and SQSTM1) related genes. It seems that the duration of physical activity and its bioenergetic cost has an important impact on the degree of increase in expression of this panel of autophagy-related genes. Anaerobic effort is more strenuous than aerobic effort and requires a higher bioenergetic investment. This may explain the stronger impact of anaerobic effort on the expression of the studied genes. This observation seems to support the protective role of autophagy proposed in prior studies
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