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Two-dimensional MXene materials perform excellently in supercapacitor applications, but self-stacking and overlap limit their applications. Constructing a reasonable layered structure by combining MXene and graphene can effectively inhibit the restacking and overlap of MXene and improve the performance of supercapacitors. In this work, we studied the energy storage performance of a conventional MXene electrode and MXene/graphene composite electrode in sulfuric acid aqueous electrolyte by molecular dynamics (MD) simulation and analyzed their energy storage mechanisms. The simulation results reveal that the MXene/graphene composite electrode showed faster charge-discharge speed and larger capacity and had more obvious advantages as a cathode. The charging process of the composite cathode can be divided into two stages. In the first stage, SO42- and H3O+ enter the electrode as a whole in a nearly 1:2 ratio, and a unique three-layer structure is formed in the graphene area, while a large number of HSO4- leaves the electrode. In the second stage, SO42- with a part of H3O+ (ratio of 2:2 to 2:3) leave the electrode, and the three-layer structure is gradually destroyed. The cooperation of these two stages leads to a particular "concave" in the total energy change of the composite cathode. The introduction of graphene has brought about changes in ion distribution, migration mechanism, and energy change, making the MXene/graphene cathode show significant advantages in energy storage. This work is of great significance for understanding the microscopic energy storage mechanism of MXene/graphene-based electrodes.
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BACKGROUND: Gastric remnant bleeding is a special case of upper gastrointestinal bleeding with certain specific disease characteristics, and some matters of transcatheter arterial embolization (TAE) for hemostasis need attention. In this study, we aimed to explore the clinical use of TAE in patients with nonvariceal gastric remnant bleeding and identify the factors influencing the clinical efficacy of these interventions. METHODS: Data were retrospectively analyzed from 42 patients for whom angiography and embolization were performed but could not be treated endoscopically or had failed endoscopic management in our department between January 2018 and January 2023 due to nonvariceal gastric remnant bleeding. We investigated the relationship between the incidence of re-bleeding and the following variables: sex, age, pre-embolization gastroscopy/contrast-enhanced computer tomography, embolization method, aortography performance, use of endoscopic titanium clips, and the presence of collateral gastric-supplying arteries. RESULTS: Forty-two patients underwent 47 interventional embolizations. Of these, 16 were positive for angiographic findings, and 26 were negative. Based on arteriography results, different embolic agents were selected, and the technical success rate was 100%. The incidence of postoperative re-bleeding was 19.1% (9/47), and the overall clinical success rate was 81.0% (34/42). Logistic regression analysis of the relationship between the incidence of early re-bleeding following embolization and the proportion of collateral gastric supply arteries revealed an odds ratio of 10.000 (p = 0.014). CONCLUSIONS: Utilizing TAE for nonvariceal gastric remnant bleeding is safe and effective. The omission of collateral gastric-supplying arteries can lead to early re-bleeding following an intervention.
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Embolización Terapéutica , Muñón Gástrico , Humanos , Estudios Retrospectivos , Hemorragia Gastrointestinal/etiología , Hemorragia Gastrointestinal/terapia , Embolización Terapéutica/efectos adversos , GastroscopíaRESUMEN
Sogatella furcifera is a destructive agricultural pest causing large threats to rice production in China and Southeast Asian countries. Despite recent breakthroughs in long-read sequencing, high quality genomic data are very limited in S. furcifera. In present study, a chromosome-level assembly of the S. furcifera genome was completed (0.64 GB), comprising 15 chromosomes covered 95.04% of the estimated genome size, along with other 624 small scaffolds making up the remaining 4.96% of the genome of S. furcifera. A total of 24,669 protein-coding genes, 1211 long noncoding RNA and 7595 circular RNA transcripts were predicted in this study. Comparative genomic analysis revealed rapidly evolved genes were associated with multiple immune-related pathways in S. furcifera. Genome resequencing of 44 individuals from 12 geographic populations revealed frequent gene flow among populations. The systemic genomic analysis will provide more insights into the understanding of the immunity and evolutionary adaptation of S. furcifera.
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Hemípteros , Metagenómica , Humanos , Animales , Genómica , China , Asia , Hemípteros/genética , CromosomasRESUMEN
Due to the unique properties of room temperature ionic liquids (RTILs), most researchers' interest in RTIL-based electrolytes in electric double-layer capacitors (EDLCs) stems from molecular simulations, which are different from experimental scientific research fields. The knowledge of RTIL-based electrolytes in EDLCs began with a supposition obtained from the results of molecular simulations of molten salts. Furthermore, experiments and simulations were promoted and developed rapidly on this topic. In some instances, the achievements of molecular simulations are ahead of even those obtained from experiments in quantity and quality. Molecular simulations offer more information on the impacts of overscreening, quasicrowding, crowding, and underscreening for RTIL-based electrolytes than experimental studies, which can be helpful in understanding the mechanisms of EDLCs. With the advancement of experimental technology, these effects have been verified by experiments. The simulation prediction of the capacitance curve was in good agreement with the experiment for pure RTILs. For complex systems, such as RTIL-solvent mixtures and RTIL mixture systems, both molecular simulations and experiments have reported that the change in capacitance curves is not monotonous with RTIL concentrations. In addition, there are some phenomena that are difficult to explain in experiments and can be well explained through molecular simulations. Finally, experiments and molecular simulations have maintained synchronous developments in recent years, and this paper discusses their relationship and reflects on their application.
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Rice stripe mosaic virus (RSMV) is an emerging pathogen which significantly reduces rice yields in the southern region of China. It is transmitted by the leafhopper Recilia dorsalis, which overwinters in rice fields. Our field investigations revealed that RSMV infection causes delayed rice heading, resulting in a large number of green diseased plants remaining in winter rice fields. This creates a favorable environment for leafhoppers and viruses to overwinter, potentially contributing to the rapid spread and epidemic of the disease. Next, we explored the mechanism by which RSMV manipulates the developmental processes of the rice plant. A rice heading-related E3 ubiquitin ligase, Heading date Associated Factor 1 (HAF1), was found to be hijacked by the RSMV-encoded P6. The impairment of HAF1 function affects the ubiquitination and degradation of downstream proteins, HEADING DATE 1 and EARLY FLOWERING3, leading to a delay in rice heading. Our results provide new insights into the development regulation-based molecular interactions between virus and plant, and highlights the importance of understanding virus-vector-plant tripartite interactions for effective disease management strategies.
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Ibuprofen (IBU), a widely used antipyretic and analgesic, has been frequently detected in various natural water systems. Advanced oxidation processes (AOPs) are effective ways to remove pollutants from water. The degradation of IBU under UV/H2O2 conditions in the presence of various kinds of natural dissolved matter was investigated using density functional theory (DFT). The eco-toxicological properties were predicted based on a quantitative structure-activity relationship (QSAR) model. The calculated results showed that two H-abstraction reactions occurring at the side chain are predominant pathways in the initial reaction. H2O, NH3, CH3OH, C2H5OH, HCOOH and CH3COOH can catalyze the H transfer in the degradation process through decreasing the energy barriers and the catalysis effects follow the order of NH3 > alcohols > acids > H2O. The catalysis effects differ under acid or alkaline conditions. The overall rate coefficient of the reaction of IBU with ËOH is calculated to be 5.04 × 109 M-1 s-1 at 298 K. IBU has harmful effects on aquatic organisms and human beings and the degradation process cannot significantly reduce its toxicity. Among all products, 2-(4-formylphenyl)propanoic acid, which is more toxic than IBU, is the most toxic with acute and chronic toxicity, developmental toxicity, mutagenicity, genotoxic carcinogenicity and irritation/corrosivity to skin. The findings in this work provide new insights into the degradation of IBU and can help to assess its environmental risks.
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Contaminantes Químicos del Agua , Purificación del Agua , Humanos , Ibuprofeno/química , Peróxido de Hidrógeno/química , Contaminantes Químicos del Agua/química , Oxidación-Reducción , Agua , Rayos UltravioletaRESUMEN
Accurately detecting student classroom behaviors in classroom videos is beneficial for analyzing students' classroom performance and consequently enhancing teaching effectiveness. To address challenges such as object density, occlusion, and multi-scale scenarios in classroom video images, this paper introduces an improved YOLOv8 classroom detection model. Firstly, by combining modules from the Res2Net and YOLOv8 network models, a novel C2f_Res2block module is proposed. This module, along with MHSA and EMA, is integrated into the YOLOv8 model. Experimental results on a classroom detection dataset demonstrate that the improved model in this paper exhibits better detection performance compared to the original YOLOv8, with an average precision (mAP@0.5) increase of 4.2%.
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The traditional Transformer model primarily employs a self-attention mechanism to capture global feature relationships, potentially overlooking local relationships within sequences and thus affecting the modeling capability of local features. For Support Vector Machine (SVM), it often requires the joint use of feature selection algorithms or model optimization methods to achieve maximum classification accuracy. Addressing the issues in both models, this paper introduces a novel network framework, CTSF, specifically designed for Industrial Internet intrusion detection. CTSF effectively addresses the limitations of traditional Transformers in extracting local features while compensating for the weaknesses of SVM. The framework comprises a pre-training component and a decision-making component. The pre-training section consists of both CNN and an enhanced Transformer, designed to capture both local and global features from input data while reducing data feature dimensions. The improved Transformer simultaneously decreases certain training parameters within CTSF, making it more suitable for the Industrial Internet environment. The classification section is composed of SVM, which receives initial classification data from the pre-training phase and determines the optimal decision boundary. The proposed framework is evaluated on an imbalanced subset of the X-IIOTID dataset, which represent Industrial Internet data. Experimental results demonstrate that with SVM using both "linear" and "rbf" kernel functions, CTSF achieves an overall accuracy of 0.98875 and effectively discriminates minor classes, showcasing the superiority of this framework.
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Phytoplasmas are uncultivable, phloem-limited, phytopathogenic bacteria that represent a major threat to agriculture worldwide. Phytoplasma membrane proteins are in direct contact with hosts and presumably play a crucial role in phytoplasma spread within the plant as well as by the insect vector. Three highly abundant types of immunodominant membrane proteins (IDP) have been identified within the phytoplasmas: immunodominant membrane protein (Imp), immunodominant membrane protein A (IdpA), and antigenic membrane protein (Amp). Although recent results indicate that Amp is involved in host specificity by interacting with host proteins such as actin, little is known about the pathogenicity of IDP in plants. In this study, we identified an antigenic membrane protein (Amp) of rice orange leaf phytoplasma (ROLP), which interacts with the actin of its vector. In addition, we generated Amp-transgenic lines of rice and expressed Amp in tobacco leaves by the potato virus X (PVX) expression system. Our results showed that the Amp of ROLP can induce the accumulation of ROLP and PVX in rice and tobacco plants, respectively. Although several studies have reported interactions between major phytoplasma antigenic membrane protein (Amp) and insect vector proteins, this example demonstrates that Amp protein can not only interact with the actin protein of its insect vector but can also directly inhibit host defense responses to promote the infection. The function of ROLP Amp provides new insights into the phytoplasma-host interaction.
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Oryza , Phytoplasma , Actinas/metabolismo , Phytoplasma/metabolismo , Oryza/metabolismo , Proteínas de la Membrana/metabolismo , Virulencia , Plantas/metabolismo , Nicotiana/metabolismo , Hojas de la Planta/metabolismo , Enfermedades de las Plantas/microbiologíaRESUMEN
Coronary angiography (CAG) as a typical imaging modality for the diagnosis of coronary diseases hasbeen widely employed in clinical practices. For CAG-based computer-aided diagnosis systems, accurate vessel segmentation plays a fundamental role. However, patients with bradycardia usually have a pacemaker which frequently interferes the vessel segmentation. In this case, the segmentation of vessels will be hard. To mitigate interferences of pacemakers and then extract main vessels more effectively in CAG images, we propose an approach. At first, a pseudo CAG (pCAG) image is generated through a part of a CAG sequence, in which the pacemaker exists. Then, a local feature descriptor is employed to register the relative location of pacemaker between the pCAG image and the target CAG image. Finally, combining the registration result and segmentation results of main vessels and pacemaker, interferences of pacemaker are removed and the segmentation of main vessels is improved. The proposed method is evaluated based on 11 CAG images with pacemakers acquired in clinical practices. An optimization ratio of the Dice coefficient is 12.04%, which demonstrates that our method can remove overlapping pacemakers and achieve the improvement of main vessel segmentation in CAG images.Our method can further become a helpful component in a CAG-based computer-aided diagnosis system, improving its diagnosis accuracy and efficiency.
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Diagnóstico por Computador , Marcapaso Artificial , Humanos , Angiografía Coronaria/métodos , Procesamiento de Imagen Asistido por Computador/métodos , AlgoritmosRESUMEN
Cap-independent translation initiation on picornavirus mRNAs is mediated by an internal ribosomal entry site (IRES) in the 5' untranslated region. The regulation of internal initiation requires the interaction of IRES-transacting factors (ITAFs) with the IRES. In this study, we identified a novel ITAF, heterogeneous nuclear ribonucleoprotein K (hnRNP K), which negatively regulates foot-and-mouth disease virus (FMDV) translation and viral replication. Further investigation revealed that the KH2 and KH3 domains of hnRNP K directly bind to domains II, III, and IV of the FMDV IRES, resulting in the inhibition of IRES-mediated translation by interfering with the recognition of another positive ITAF, polypyrimidine tract-binding protein (PTB). Conversely, hnRNP K-mediated inhibition was antagonized by the viral 3C protease through the cleavage of hnRNP K at the Glu-364 residue during FMDV infection. Interestingly, the N-terminal cleavage product, hnRNP K1-364, retained partial inhibitory effects on IRES activity, whereas the C-terminal cleavage product, hnRNP K364-465, became a positive regulator of FMDV replication. Our findings expand the current understanding of virus-host interactions concerning viral recruitment and the modulation of ITAFs, providing new insights into translational control during viral infection.IMPORTANCE The translation of picornaviral genome RNA mediated by the internal ribosomal entry site (IRES) is a crucial step for virus infections. Virus-host interactions play a critical role in the regulation of IRES-dependent translation, but the regulatory mechanism remains largely unknown. In this study, we identified an ITAF, hnRNP K, that negatively regulates FMDV replication by inhibiting viral IRES-mediated translation. In addition, we describe a novel translational regulation mechanism involving the proteolytic cleavage of hnRNP K by FMDV protease 3C. The cleavage of hnRNP K yields two cleavage products with opposite functions: the cleavage product hnRNP K1-364 retains a partial inhibitory effect on IRES activity, and the cleavage product hnRNP K364-465 becomes a positive regulator of FMDV replication. Our findings shed light on the effect of a novel ITAF on the translational regulation of picornavirus and provide new insights into translational control during viral infection.
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Cisteína Endopeptidasas/metabolismo , Virus de la Fiebre Aftosa/fisiología , Ribonucleoproteína Heterogénea-Nuclear Grupo K/metabolismo , Sitios Internos de Entrada al Ribosoma/fisiología , Transactivadores/metabolismo , Proteínas Virales/metabolismo , Replicación Viral/fisiología , Proteasas Virales 3C , Animales , Línea Celular , Cricetinae , Virus de la Fiebre Aftosa/genética , Regulación Viral de la Expresión Génica , Células HEK293 , Humanos , Proteína de Unión al Tracto de Polipirimidina , ARN Mensajero , Proteínas Virales/genéticaRESUMEN
Upon infection, the highly structured 5' untranslated region (5' UTR) of picornavirus is involved in viral protein translation and RNA synthesis. As a critical element in the 5' UTR, the internal ribosome entry site (IRES) binds to various cellular proteins to function in the processes of picornavirus replication. Foot-and-mouth disease virus (FMDV) is an important member in the family Picornaviridae, and its 5' UTR contains a functional IRES element. In this study, the cellular heterogeneous nuclear ribonucleoprotein L (hnRNP L) was identified as an IRES-binding protein for FMDV by biotinylated RNA pulldown assays, mass spectrometry (MS) analysis, and determination of hnRNP L-IRES interaction regions. Further, we found that hnRNP L inhibited the growth of FMDV through binding to the viral IRES and that the inhibitory effect of hnRNP L on FMDV growth was not due to FMDV IRES-mediated translation, but to influence on viral RNA synthesis. Finally, hnRNP L was demonstrated to coimmunoprecipitate with RNA-dependent RNA polymerase (3Dpol) in an FMDV RNA-dependent manner in the infected cells. Thus, our results suggest that hnRNP L, as a critical IRES-binding protein, negatively regulates FMDV replication by inhibiting viral RNA synthesis, possibly by remaining in the replication complex.IMPORTANCE Picornaviruses, as a large family of human and animal pathogens, cause a bewildering array of disease syndromes. Many host factors are implicated in the pathogenesis of these viruses, and some proteins interact with the viral IRES elements to affect function. Here, we report for the first time that cellular hnRNP L specifically interacts with the IRES of the picornavirus FMDV and negatively regulates FMDV replication through inhibiting viral RNA synthesis. Further, our results showed that hnRNP L coimmunoprecipitates with FMDV 3Dpol in a viral RNA-dependent manner, suggesting that it may remain in the replication complex to function. The data presented here would facilitate further understanding of virus-host interactions and the pathogenesis of picornavirus infections.
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Regiones no Traducidas 5' , Virus de la Fiebre Aftosa/fisiología , Ribonucleoproteína Heterogénea-Nuclear Grupo L/metabolismo , Sitios Internos de Entrada al Ribosoma/fisiología , ARN Viral/biosíntesis , Replicación Viral/fisiología , Animales , Línea Celular , Virus de la Fiebre Aftosa/genética , Regulación Viral de la Expresión Génica , Técnicas de Inactivación de Genes , Células HEK293 , Ribonucleoproteína Heterogénea-Nuclear Grupo L/genética , Interacciones Microbiota-Huesped/fisiología , Humanos , Inmunoprecipitación , Unión Proteica , ARN Viral/genética , TranscriptomaRESUMEN
The low fidelity of foot-and-mouth disease virus (FMDV) RNA-dependent RNA polymerase allows FMDV to exhibit high genetic diversity. Previously, we showed that the genetic diversity of FMDV plays an important role in virulence in suckling mice. Here, we mutated the amino acid residue Phe257, located in the finger domain of FMDV polymerase and conserved across FMDV serotypes, to a cysteine (F257C) to study the relationship between viral genetic diversity, virulence, and transmissibility in natural hosts. The single amino acid substitution in FMDV polymerase resulted in a high-fidelity virus variant, rF257C, with growth kinetics indistinguishable from those of wild-type (WT) virus in cell culture, but it displayed smaller plaques and impaired fitness in direct competition assays. Furthermore, we found that rF257C was attenuated in vivo in both suckling mice and pigs (one of its natural hosts). Importantly, contact exposure experiments showed that the rF257C virus exhibited reduced transmissibility compared to that of wild-type FMDV in the porcine model. This study provides evidence that FMDV genetic diversity is important for viral virulence and transmissibility in susceptible animals. Given that type O FMDV exhibits the highest genetic diversity among all seven serotypes of FMDV, we propose that the lower polymerase fidelity of the type O FMDV could contribute to its dominance worldwide.IMPORTANCE Among the seven serotypes of FMDV, serotype O FMDV have the broadest distribution worldwide, which could be due to their high virulence and transmissibility induced by high genetic diversity. In this paper, we generated a single amino acid substitution FMDV variant with a high-fidelity polymerase associated with viral fitness, virulence, and transmissibility in a natural host. The results highlight that maintenance of viral population diversity is essential for interhost viral spread. This study provides evidence that higher genetic diversity of type O FMDV could increase both virulence and transmissibility, thus leading to their dominance in the global epidemic.
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Virus de la Fiebre Aftosa/patogenicidad , Fiebre Aftosa/virología , ARN Polimerasa Dependiente del ARN/fisiología , Proteínas no Estructurales Virales/fisiología , Animales , Línea Celular , Cricetinae , Virus de la Fiebre Aftosa/enzimología , Virus de la Fiebre Aftosa/genética , Aptitud Genética , Variación Genética , Ratones , Mutación , Fenotipo , ARN Polimerasa Dependiente del ARN/genética , Porcinos , Proteínas no Estructurales Virales/genética , VirulenciaRESUMEN
Foot-and-mouth disease (FMD), which is caused by FMD virus (FMDV), remains a major plague among cloven-hoofed animals worldwide, and its outbreak often has disastrous socioeconomic consequences. A live-attenuated FMDV vaccine will greatly facilitate the global control and eradication of FMD, but a safe and effective attenuated FMDV vaccine has not yet been successfully developed. Here, we found that the internal ribosome entry site (IRES) element in the viral genome is a critical virulence determinant of FMDV, and a nucleotide substitution of cytosine (C) for guanine (G) at position 351 of the IRES endows FMDV with temperature-sensitive and attenuation (ts&att) phenotypes. Furthermore, we demonstrated that the C351G mutation of IRES causes a temperature-dependent translation defect by impairing its binding to cellular pyrimidine tract-binding protein (PTB), resulting in the ts&att phenotypes of FMDV. Natural hosts inoculated with viruses carrying the IRES C351G mutation showed no clinical signs, viremia, virus excretion, or viral transmission but still produced a potent neutralizing antibody response that provided complete protection. Importantly, the IRES C351G mutation is a universal determinant of the ts&att phenotypes of different FMDV strains, and the C351G mutant was incapable of reversion to virulence during in vitro and in vivo passages. Collectively, our findings suggested that manipulation of the IRES, especially its C351G mutation, may serve as a feasible strategy to develop live-attenuated FMDV vaccines.IMPORTANCE The World Organization for Animal Health has called for global control and eradication of foot-and-mouth disease (FMD), the most economically and socially devastating disease affecting animal husbandry worldwide. Live-attenuated vaccines are considered the most effective strategy for prevention, control, and eradication of infectious diseases due to their capacity to induce potent and long-lasting protective immunity. However, efforts to develop FMD virus (FMDV) live-attenuated vaccines have achieved only limited success. Here, by structure-function study of the FMDV internal ribosome entry site (IRES), we find that the C351 mutation of the IRES confers FMDV with an ideal temperature-sensitive attenuation phenotype by decreasing its interaction with cellular pyrimidine tract-binding protein (PTB) to cause IRES-mediated temperature-dependent translation defects. The temperature-sensitive attenuated strains generated by manipulation of the IRES address the challenges of FMDV attenuation differences among various livestock species and immunogenicity maintenance encountered previously, and this strategy can be applied to other viruses with an IRES to rationally design and develop live-attenuated vaccines.
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Virus de la Fiebre Aftosa/genética , Sitios Internos de Entrada al Ribosoma/genética , Animales , Anticuerpos Neutralizantes/metabolismo , Bovinos , Femenino , Fiebre Aftosa/virología , Virus de la Fiebre Aftosa/metabolismo , Virus de la Fiebre Aftosa/patogenicidad , Regulación Viral de la Expresión Génica/genética , Sitios Internos de Entrada al Ribosoma/fisiología , Masculino , Ratones , Ratones Endogámicos BALB C , Mutación/genética , Ribosomas/genética , Porcinos , Vacunas Atenuadas , Virulencia/genética , Replicación Viral/genéticaRESUMEN
Viral diseases severely affect crop yield and quality, thereby threatening global food security. Genetic improvement of plant virus resistance is essential for sustainable agriculture. In the last decades, several modern technologies were applied in plant antiviral engineering. Here we summarized breakthroughs of the two major antiviral strategies, RNA silencing and genome editing. RNA silencing strategy has been used in antiviral breeding for more than thirty years, and many crops engineered to stably express small RNAs targeting various viruses have been approved for commercial release. Genome editing technology has emerged in the past decade, especially CRISPR/Cas, which provides new methods for genetic improvement of plant virus resistance and accelerates resistance breeding. Finally, we discuss the potential of these technologies for breeding crops, and the challenges and solutions they may face in the future.
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Productos Agrícolas , Resistencia a la Enfermedad , Edición Génica , Sistemas CRISPR-Cas , Productos Agrícolas/genética , Productos Agrícolas/virología , Resistencia a la Enfermedad/genética , Genoma de Planta/genética , Virus de Plantas , Interferencia de ARNRESUMEN
Senecavirus A (SVA) is a reemerging virus, and recent evidence has emphasized the importance of SVA recombination in vivo on virus evolution. In this study, we report the development of an infectious cDNA clone for the SVA/HLJ/CHA/2016 strain. We used this strain to develop a reporter virus expressing enhanced green fluorescent protein (eGFP), which we then used to screen for a recombination-deficient SVA by an eGFP retention assay. Sequencing of the virus that retained the eGFP following passage allowed us to identify the nonsynonymous mutations (S460L alone and I212V-S460L in combination) in the RNA-dependent RNA polymerase (RdRp) region of the genome. We developed a Senecavirus-specific cell culture-based recombination assay, which we used to elucidate the role of RdRp in SVA recombination. Our results demonstrate that these two polymerase variants (S460L and I212/S460L) have reduced recombination capacity. These results indicate that the RdRp plays a central role in SVA replicative recombination. Notably, our results showed that the two recombination-deficient variants have higher replication fidelity than the wild type (WT) and display decreased ribavirin sensitivity compared to the WT. In addition, these two mutants exhibited significantly increased fitness in vitro compared to the WT. These results demonstrate that recombination and mutation rates are intimately linked. Our results have important implications for understanding the crucial role of the RdRp in virus recombination and fitness, especially in the molecular mechanisms of SVA evolution and pathogenicity.IMPORTANCE Recent evidence has emphasized the importance of SVA recombination on virus evolution in vivo We describe the first assays to study Senecavirus A recombination. The results show that the RNA-dependent RNA polymerase plays a crucial role in recombination and that recombination can impact the fitness of SVA in cell culture. Further, SVA polymerase fidelity is closely related to recombination efficiency. The results provide key insights into the role of recombination in positive-strand RNA viruses.
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Picornaviridae/genética , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/genética , ARN , Recombinación Genética , Animales , Antivirales/farmacología , Línea Celular , ADN Complementario , Farmacorresistencia Viral/efectos de los fármacos , Regulación Viral de la Expresión Génica , Genotipo , Modelos Moleculares , Mutación , Tasa de Mutación , Fenotipo , Picornaviridae/efectos de los fármacos , Conformación Proteica , ARN Polimerasa Dependiente del ARN/química , Ribavirina/farmacología , Análisis de SecuenciaRESUMEN
Rice orange leaf disease (ROLD), caused by rice orange leaf phytoplasma (ROLP), is transmitted by leafhopper vectors Recilia dorsalis and Nephotettix cinticeps. ROLD severely devastates rice production in Asia. Accurate detection of the pathogen is important for disease management. Current nested polymerase chain reaction (nested PCR) method using phytoplasma universal primers is widely used to detect phytoplasmas; however, it has shortcoming of inconvenience and inaccuracy, for it needs two round of PCR reactions and could produce false positive results due to nontarget amplification. In this study, we developed a PCR assay using a set of primers designed based on the ROLP genome sequence to amplify house-keeping gene FtsH-1 in rice and leafhopper vector samples. This method is simple and rapid, and its sensitivity up to 10 pg/µl of total ROLP DNA. It also minimizes the false positive problem produced by nested PCR. This method was used to survey the geographic distribution of ROLD in southern China from 2016 to 2018. The results showed that the distribution areas and vector carrying rate of ROLD had gradually increased.
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Oryza , Phytoplasma , Animales , China , Enfermedades de las Plantas , Reacción en Cadena de la PolimerasaRESUMEN
BACKGROUND: Rice stripe mosaic virus (RSMV) is a tentative new Cytorhabdovirus species in family Rhabdoviridae transmitted by the leafhopper Recilia dorsalis. Although the virus was first detected in southern China in 2015, few studies have investigated rice symptoms and yield losses caused by RSMV infection. METHODS: In this study, we observed and systematically compared symptoms of three virally infected, representative varieties of indica, hybrid and japonica rice and determined the yield parameters of the artificially inoculated plants. RESULTS: The three RSMV-infected cultivated rice varieties exhibited slight dwarfing, striped mosaicism, stiff, crinkled or even twisted leaves, an increased number of tillers, delayed heading, cluster-shaped shortening of panicles and mostly unfilled grains. Slight differences in symptom occurrence time were observed under different environmental conditions. For example, mosaic symptoms appeared earlier and crinkling symptoms appeared later, with both symptoms later receding in some infected plants. Yield losses due to RSMV also differed among varieties. The most serious yield reduction was experienced by indica rice (cv. Meixiangzhan), followed by hybrid indica rice (cv. Wuyou 1179) and then japonica (cv. Nipponbare). Single panicle weight, seed setting rate and 1000-kernel weight were reduced in the three infected varieties compared with healthy plants-by 85.42, 94.85 and 31.56% in Meixiangzhan; 52.43, 53.06 and 25.65% in Wuyou 1179 and 25.53, 49.32 and 23.86% in Nipponbare, respectively. CONCLUSIONS: Our findings contribute basic data for field investigations, formulation of prevention and control strategies and further study of the pathogenesis of RSMV.
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Oryza/crecimiento & desarrollo , Oryza/virología , Enfermedades de las Plantas/virología , Infecciones por Rhabdoviridae/virología , Rhabdoviridae/crecimiento & desarrollo , Quimera/anatomía & histología , Quimera/crecimiento & desarrollo , Quimera/virología , China , Oryza/anatomía & histologíaRESUMEN
Recently, CRISPR-Cas (clustered, regularly interspaced short palindromic repeats-CRISPR-associated proteins) system has been used to produce plants resistant to DNA virus infections. However, there is no RNA virus control method in plants that uses CRISPR-Cas system to target the viral genome directly. Here, we reprogrammed the CRISPR-Cas9 system from Francisella novicida to confer molecular immunity against RNA viruses in Nicotiana benthamiana and Arabidopsis plants. Plants expressing FnCas9 and sgRNA specific for the cucumber mosaic virus (CMV) or tobacco mosaic virus (TMV) exhibited significantly attenuated virus infection symptoms and reduced viral RNA accumulation. Furthermore, in the transgenic virus-targeting plants, the resistance was inheritable and the progenies showed significantly less virus accumulation. These data reveal that the CRISPR/Cas9 system can be used to produce plant that stable resistant to RNA viruses, thereby broadening the use of such technology for virus control in agricultural field.
Asunto(s)
Francisella/genética , Enfermedades de las Plantas/virología , Plantas Modificadas Genéticamente/inmunología , Plantas Modificadas Genéticamente/virología , Virus ARN/patogenicidad , Sistemas CRISPR-Cas , Enfermedades de las Plantas/inmunología , Plantas Modificadas Genéticamente/metabolismo , Virus ARN/inmunologíaRESUMEN
BACKGROUND: Virus-derived siRNAs (vsiRNAs)-mediated RNA silencing plays important roles in interaction between plant viruses and their hosts. Southern rice black-streaked dwarf virus (SRBSDV) is a newly emerged devastating rice reovirus with ten dsRNA genomic segments. The characteristics of SRBSDV-derived siRNAs and their biological implications in SRBSDV-rice interaction remain unexplored. METHODS: VsiRNAs profiling from SRBSDV-infected rice samples was done via small RNA deep sequencing. The putative rice targets of abundantly expressed vsiRNAs were bioinformatically predicted and subjected to functional annotation. Differential expression analysis of rice targets and RNA silencing components between infected and healthy samples was done using RT-qPCR. RESULTS: The vsiRNA was barely detectable at 14 days post infection (dpi) but abundantly present along with elevated expression level of the viral genome at 28 dpi. From the 28-dpi sample, 70,878 reads of 18 ~ 30-nt vsiRNAs were recognized (which mostly were 21-nt and 22-nt), covering 75 ~ 91% of the length of the ten genomic segments respectively. 86% of the vsiRNAs had a <50% GC content and 79% of them were 5'-uridylated or adenylated. The production of vsiRNAs had no strand polarity but varied among segment origins. Each segment had a few hotspot regions where vsiRNAs of high abundance were produced. 151 most abundant vsiRNAs were predicted to target 844 rice genes, including several types of host resistance or pathogenesis related genes encoding F-box/LRR proteins, receptor-like protein kinases, universal stress proteins, tobamovirus multiplication proteins, and RNA silencing components OsDCL2a and OsAGO17 respectively, some of which showed down regulation in infected plants in RT-qPCR. GO and KEGG classification showed that a majority of the predicted targets were related to cell parts and cellular processes and involved in carbohydrate metabolism, translation, and signal transduction. The silencing component genes OsDCL2a, OsDCL2b, OsDCL4, and OsAGO18 were down regulated, while OsAGO1d, OsAGO2, OsRDR1 and OsRDR6 were up regulated, significantly, upon SRBSDV infection. CONCLUSIONS: SRBSDV can regulate the expression of rice RNA silencing pathway components and the virus might compromise host defense and influence host pathogenesis via siRNA pathways.