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Abstract: Hypersensitive response (HR)-conferred resistance to viral infection restricts the virus spread and is accompanied by the induction of cell death, manifested as the formation of necrotic lesions. While it is known that salicylic acid is the key component in the orchestration of the events restricting viral spread in HR, the exact function of the cell death in resistance is still unknown. We show that potato virus Y (PVY) can be detected outside the cell death zone in Ny-1-mediated HR in potato plants (cv. Rywal), observed as individual infected cells or small clusters of infected cells outside the cell death zone. By exploiting the features of temperature dependent Ny-1-mediated resistance, we confirmed that the cells at the border of the cell death zone are alive and harbor viable PVY that is able to reinitiate infection. To get additional insights into this phenomenon we further studied the dynamics of both cell death zone expansion and occurrence of viral infected cell islands outside it. We compared the response of Rywal plants to their transgenic counterparts, impaired in SA accumulation (NahG-Rywal), where the lesions occur but the spread of the virus is not restricted. We show that the virus is detected outside the cell death zone in all lesion developmental stages of HR lesions. We also measured the dynamics of lesions expansion in both genotypes. We show that while rapid lesion expansion is observed in SA-depleted plants, virus spread is even faster. On the other hand the majority of analyzed lesions slowly expand also in HR-conferred resistance opening the possibility that the infected cells are eventually engulfed by cell death zone. Taken altogether, we suggest that the HR cell death is separated from the resistance mechanisms which lead to PVY restriction in Ny-1 genetic background. We propose that HR should be regarded as a process where the dynamics of events is crucial for effectiveness of viral arrest albeit the exact mechanism conferring this resistance remains unknown.
Keywords: potato virus Y, salicylic acid, hypersensitive response, programmed cell death, callose deposits, necrotic lesion
Published in DiRROS: 24.07.2024; Views: 266; Downloads: 236
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Abstract: Cell lines which are currently used in genotoxicity tests lack enzymes which activate/detoxify mutagens. Therefore, rodent-derived liver preparations are used which reflect their metabolism in humans only partly; as a consequence misleading results are often obtained. Previous findings suggest that certain liver cell lines express phase I/II enzymes and detect promutagens without activation; however, their use is hampered by different shortcomings. The aim of this study was the identification of a suitable cell line. The sensitivity of twelve hepatic cell lines was investigated in single cell gel electrophoresis assays. Furthermore, characteristics of these lines were studied which are relevant for their use in genotoxicity assays (mitotic activity, p53 status, chromosome number, and stability). Three lines (HuH6, HCC1.2, and HepG2) detected representatives of five classes of promutagens, namely, IQ and PhIP (HAAs), B(a)P (PAH), NDMA (nitrosamine), and AFB1 (aflatoxin), and were sensitive towards reactive oxygen species (ROS). In contrast, the commercially available line HepaRG, postulated to be a surrogate for hepatocytes and an ideal tool for mutagenicity tests, did not detect IQ and was relatively insensitive towards ROS. All other lines failed to detect two or more compounds. HCC1.2 cells have a high and unstable chromosome number and mutated p53, these features distract from its use in routine screening. HepG2 was frequently employed in earlier studies, but pronounced inter-laboratory variations were observed. HuH6 was never used in genotoxicity experiments and is highly promising, it has a stable karyotype and we demonstrated that the results of genotoxicity experiments are reproducible.
Keywords: hepatic cell lines, p53, comet assay, genotoxicity
Published in DiRROS: 24.07.2024; Views: 286; Downloads: 189
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Abstract: Sustainable water basin management requires characterization of flow regime in river networks impacted by anthropogenic pressures. Flow regime in ungauged catchments under current, future, or natural conditions can be assessed with hydrological models. Developing hydrological models is, however, resource demanding such that decision makers might revert to models that have been developed for other purposes and are made available to them (‘off-the-shelf’ models). In this study, the impact of epistemic uncertainty of flow regime indicators on flow-ecological assessment was assessed at selected stations with drainage areas ranging from about 400 to almost 90,000 km2 in four South European basins (Adige, Ebro, Evrotas and Sava). For each basin, at least two models were employed. Models differed in structure, data input, spatio-temporal resolution, and calibration strategy, reflecting the variety of conditions and purposes for which they were initially developed. The uncertainty of modelled flow regime was assessed by comparing the modelled hydrologic indicators of magnitude, timing, duration, frequency and rate of change to those obtained from observed flow. The results showed that modelled flow magnitude indicators at medium and high flows were generally reliable, whereas indicators for flow timing, duration, and rate of change were affected by large uncertainties, with correlation coefficients mostly below 0.50. These findings mirror uncertainty in flow regime indicators assessed with other methods, including from measured streamflow. The large indicator uncertainty may significantly affect assessment of ecological status in freshwater systems, particularly in ungauged catchments. Finally, flow-ecological assessments proved very sensitive to reference flow regime (i.e., without anthropogenic pressures). Model simulations could not adequately capture flow regime in the reference sites comprised in this study. The lack of reliable reference conditions may seriously hamper flow-ecological assessments. This study shows the pressing need for improving assessment of natural flow regime at pan-European scale.
Keywords: flow regime, ecohydrological assessment, SWAT, mHM, lisflood, HYPERstream
Published in DiRROS: 24.07.2024; Views: 326; Downloads: 173
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Keywords: arhitekturna zgodovina
Published in DiRROS: 24.07.2024; Views: 284; Downloads: 244
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Abstract: Instances of sexual size dimorphism (SSD) provide the context for rigorous tests of biological rules of size evolution, such as Cope’s rule (phyletic size increase), Rensch’s rule (allometric patterns of male and female size), as well as male and female body size optima. In certain spider groups, such as the golden orbweavers (Nephilidae), extreme female-biased SSD (eSSD, female:male body length 2) is the norm. Nephilid genera construct webs of exaggerated proportions, which can be aerial, arboricolous, or intermediate (hybrid). First, we established the backbone phylogeny of Nephilidae using 367 anchored hybrid enrichment markers, then combined these data with classical markers for a reference species-level phylogeny. Second, we used the phylogeny to test Cope and Rensch’s rules, sex specific size optima, and the coevolution of web size, type, and features with female and male body size and their ratio, SSD. Male, but not female, size increases significantly over time, and refutes Cope’s rule. Allometric analyses reject the converse, Rensch’s rule. Male and female body sizes are uncorrelated. Female size evolution is random, but males evolve toward an optimum size (3.2–4.9 mm). Overall, female body size correlates positively with absolute web size. However, intermediate sized females build the largest webs (of the hybrid type), giant female Nephila and Trichonephila build smaller webs (of the aerial type), and the smallest females build the smallest webs (of the arboricolous type). We propose taxonomic changes based on the criteria of clade age, monophyly and exclusivity, classification information content, and diagnosability. Spider families, as currently defined, tend to be between 37 million years old and 98 million years old, and Nephilidae is estimated at 133 Ma (97–146), thus deserving family status. We, therefore, resurrect the family Nephilidae Simon 1894 that contains Clitaetra Simon 1889, the Cretaceous GeratonephilaPoinar and Buckley (2012), Herennia Thorell 1877, IndoetraKuntner 2006, new rank, Nephila Leach 1815, Nephilengys L. Koch 1872, Nephilingis Kuntner 2013, Palaeonephila Wunderlich 2004 from Tertiary Baltic amber, and TrichonephilaDahl 1911, new rank. We propose the new clade Orbipurae to contain Araneidae Clerck 1757, Phonognathidae Simon 1894, new rank, and Nephilidae. Nephilid female gigantism is a phylogenetically ancient phenotype (over 100 Ma), as is eSSD, though their magnitudes vary by lineage.
Keywords: spiders, evolution of sexual size dimorphism, golden orbweavers, analyses
Published in DiRROS: 24.07.2024; Views: 482; Downloads: 229
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