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Abstract: The emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants poses a challenge to current therapies and emphasises the need for targets that are less susceptible to mutation. The SARS-CoV-2 envelope protein (Epro) is a highly conserved viroporin and is of central importance to the viral life cycle, yet it remains underexplored as a therapeutic target. In this study, we have identified and characterised a novel lead candidate (LC) − (S)-N-(2-((1S,3S,5S,7S)-adamantan-2-yl)ethyl)-2-(butylamino)-3-(1H-indol-3-yl)propanamide − for inhibition of the SARS-CoV-2 Epro ion channel using combined in silico and in vitro approaches. Molecular dynamics (MD) simulations showed that LC forms stable complexes at the N-terminal vestibule, with key interactions at GLU8, THR9, THR11, ASN15, and LEU18, and a calculated binding affinity higher than that of the reference compound rimantadine within the applied MD/molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) framework. Results of in vitro experiments indicated that LC inhibits the model coronavirus murine hepatitis virus at late stages of the viral cycle. Activity under co-treatment conditions further suggests a direct virucidal effect or interference with early entry stages; an EC50 of approximately 12 μM was within the same micromolar range as those observed for the reference Epro inhibitors 5-(N,N-hexamethylene)amiloride and rimantadine. Quantitative PCR experiments showed delayed RNA replication in LC-treated infected cells, while light and transmission electron microscopy displayed the reduced release of virions and prevention of cell lysis. These results emphasise the central role of the Epro ion channel in the coronavirus life cycle and present LC as a promising candidate for the further development of novel coronavirus inhibitors.
Keywords: SARS-CoV-2 envelope protein, viroporin, ion channel blockers, molecular docking, molecular dynamics simulations, drug discovery, murine hepatitis virus, in vitro
Published in DiRROS: 11.06.2026; Views: 337; Downloads: 360
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Keywords: allelopathy, Bohemian knotweed, Japanese knotweed, oxidative stress, root hair, transcriptomics
Published in DiRROS: 26.09.2025; Views: 710; Downloads: 355
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Abstract: Potato (Solanum tuberosum) is highly water and space efficient but susceptible to abiotic stresses such as heat, drought, and flooding, which are severely exacerbated by climate change. Our understanding of crop acclimation to abiotic stress, however, remains limited. Here, we present a comprehensive molecular and physiological high-throughput profiling of potato (Solanum tuberosum, cv. Désirée) under heat, drought, and waterlogging applied as single stresses or in combinations designed to mimic realistic future scenarios. Stress responses were monitored via daily phenotyping and multi-omics analyses of leaf samples comprising proteomics, targeted transcriptomics, metabolomics, and hormonomics at several timepoints during and after stress treatments. Additionally, critical metabolites of tuber samples were analyzed at the end of the stress period. We performed integrative multi-omics data analysis using a bioinformatic pipeline that we established based on machine learning and knowledge networks. Waterlogging produced the most immediate and dramatic effects on potato plants, interestingly activating ABA responses similar to drought stress. In addition, we observed distinct stress signatures at multiple molecular levels in response to heat or drought and to a combination of both. In response to all treatments, we found a downregulation of photosynthesis at different molecular levels, an accumulation of minor amino acids, and diverse stress-induced hormones. Our integrative multi-omics analysis provides global insights into plant stress responses, facilitating improved breeding strategies toward climate-adapted potato varieties.
Keywords: potato, Solanum tuberosum, abiotic stress responses, heat, drought, waterlogging, multi-omics, integrative omics, adaptomics, panomics
Published in DiRROS: 14.04.2025; Views: 1512; Downloads: 1267
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Abstract: Glucanases are enzymes regulating the size exclusion limit and permeability of plasmodesmata and play a role in biotic stress. In plant genomes, they are encoded as relatively large gene families divided into four classes. Most studies of plant virus interactions have focused on glucanases from classes I and II. In our study, we have evaluated the role of the β-1,3-glucanase class III (Glu-III) gene in the potato–potato virus YNTN (PVYNTN) interaction and implemented the findings to plant biotechnology application. Potato cultivars Désirée and Santé, which are tolerant and extremely resistant to PVYNTN, respectively, were stably transformed with Agrobacterium tumefaciens harbouring constructs for Glu-III overexpression. Localization of Glu-III protein in patches within the cell wall was determined by tagging the Glu-III protein with green fluorescent protein. Transgenic and non-transgenic plants were challenged with PVYNTN and its multiplication and spreading was followed. Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé. In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested. The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.
Keywords: plant biotechnology, plant-virus interaction, potato virus Y, agroinfiltration, beta-1, 3-glucanase
Published in DiRROS: 04.03.2025; Views: 1258; Downloads: 1296
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Abstract: Abscission occurs specifically in the abscission zone (AZ) tissue as a natural stage of plant development. Previously, we observed delay of tomato (Solanum lycopersicum) leaf abscission when the LX ribonuclease (LX) was inhibited. The known association between LX expression and programmed cell death (PCD) suggested involvement of PCD in abscission. In this study, hallmarks of PCD were identified in the tomato leaf and flower AZs during the late stage of abscission. These included loss of cell viability, altered nuclear morphology, DNA fragmentation, elevated levels of reactive oxygen species and enzymatic activities, and expression of PCD-associated genes. Overexpression of antiapoptotic proteins resulted in retarded abscission, indicating PCD requirement. PCD, LX, and nuclease gene expression were visualized primarily in the AZ distal tissue, demonstrating an asymmetry between the two AZ sides. Asymmetric expression was observed for genes associated with cell wall hydrolysis, leading to AZ, or associated with ethylene biosynthesis, which induces abscission. These results suggest that different abscission-related processes occur asymmetrically between the AZ proximal and distal sides. Taken together, our findings identify PCD as a key mechanism that occurs asymmetrically during normal progression of abscission and suggest an important role for LX in this PCD process.
Keywords: plant cell, cell death, abscission
Published in DiRROS: 05.08.2024; Views: 1421; Downloads: 1096
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