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Background: Plant viral diseases, namely 'plant cancer', are extremely difficult to control. Even worse, few antiviral agents can effectively control and totally block viral infection. There is an urgent need to explore and discover novel agrochemicals with high activity and a unique mode of action to manage these refractory diseases. Results: Forty‐one new phenothiazine derivatives were prepared and their inhibitory activity against tobacco mosaic virus (TMV) was assessed. Compound A8 had the highest protective activity against TMV, with a half‐maximal effective concentration (EC50) of 115.67 μg/mL, which was significantly better than that of the positive controls ningnanmycin (271.28 μg/mL) and ribavirin (557.47 μg/mL). Biochemical assays demonstrated that compound A8 could inhibit TMV replication by disrupting TMV self‐assembly, but also enabled the tobacco plant to enhance its defense potency by increasing the activities of various defense enzymes. Conclusion: In this study, novel phenothiazine derivatives were elaborately fabricated and showed remarkable anti‐TMV behavior that possessed the dual‐action mechanisms of inhibiting TMV assembly and invoking the defense responses of tobacco plants. Moreover, new agrochemical alternatives based on phenothiazine were assessed for their antiviral activities and showed extended agricultural application. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Tobacco Mosaic Virus (TMV) and Potato Virus Y (PVY) pose significant threats to crop production. Non-destructive and accurate surveillance is crucial to effective disease control. In this study, we propose the adoption of hyperspectral and machine learning technologies to discern the type and severity of tobacco leaves affected by PVY and TMV infection. Initially, we applied three preprocessing methods - Multivariate Scattering Correction (MSC), Standard Normal Variate (SNV), and Savitzky-Golay smoothing filter (SavGol) - to corrected the leaf full-length spectral sheet data (350-2500nm). Subsequently, we employed two classifiers, support vector machine (SVM) and random forest (RF), to establish supervised classification models, including binary classification models (healthy/diseased leaves or PVY/TMV infected leaves) and six-class classification models (healthy and various severity levels of diseased leaves). Based on the core evaluation index, our models achieved accuracies in the range of 91-100% in the binary classification. In general, SVM demonstrated superior performance compared to RF in distinguishing leaves infected with PVY and TMV. Different combinations of preprocessing methods and classifiers have distinct capabilities in the six-class classification. Notably, SavGol united with SVM gave an excellent performance in the identification of different PVY severity levels with 98.1% average precision, and also achieved a high recognition rate (96.2%) in the different TMV severity level classifications. The results further highlighted that the effective wavelengths captured by SVM, 700nm and 1800nm, would be valuable for estimating disease severity levels. Our study underscores the efficacy of integrating hyperspectral technology and machine learning, showcasing their potential for accurate and non-destructive monitoring of plant viral diseases. [ABSTRACT FROM AUTHOR]

Since many waterborne diseases are caused by human pathogenic viruses, virus monitoring of drinking water (DW) and DW sources is crucial for public health. Therefore, the aim of this review was to describe the occurrence of human pathogenic viruses in DW and DW sources; the occurrence of two viruses proposed as novel indicators of human faecal contamination (Pepper mild mottle virus and Tobacco mosaic virus) was also reported. This research was focused on articles that assessed viral occurrence using molecular methods in the surface water used for DW production (SW-D), groundwater used for DW production (GW-D), DW and bottled-DW (BW). A total of 1544 studies published in the last 10 years were analysed, and 79 were ultimately included. In considering the detection methods, filtration is the most common concentration technique, while quantitative polymerase chain reaction is the most common quantification technique. Regarding virus occurrence in SW-D, GW-D, and DW, high percentages of positive samples were reported for adenovirus, polyomavirus and Pepper mild mottle virus. Viral genomes were frequently detected in SW-D and rarely in GW-D, suggesting that GW-D may be a safe DW source. Viral genomes were also detected in DW, posing a possible threat to human health. The lowest percentages of positive samples were found in Europe, while the highest were found in Asia and South America. Only three articles assessed viral occurrence in BW. This review highlights the lack of method standardization and the need for legislation updates. [Display omitted] [ABSTRACT FROM AUTHOR]

Mixed infection between two or more begomoviruses is commonly found in tomato fields and can affect disease outcomes by increasing symptom severity and viral accumulation compared with single infection. Viruses that affect tomato include tomato severe rugose virus (ToSRV) and tomato rugose mosaic virus (ToRMV). Previous work showed that in mixed infection, ToRMV negatively affects the infectivity and accumulation of ToSRV. ToSRV and ToRMV share a high degree of sequence identity, including cis-elements in the common region (CR) and their specific recognition sites (iteron-related domain, IRD) within the Rep gene. Here, we investigated if divergent sites in the CR and IRD are involved in the interaction between these two begomoviruses. ToSRV clones were constructed containing the same nucleotides as ToRMV in the CR (ToSRV-A(ToR:CR)), IRD (ToSRV-A(ToR:IRD)) and in both regions (ToSRV-A(ToR:CR+IRD)). When plants were co-inoculated with ToRMV and ToSRV-A(ToR:IRD), the infectivity and accumulation of ToSRV were negatively affected. In mixed inoculation of ToRMV with ToSRV-A(ToR:CR), high infectivity of both viruses and high DNA accumulation of ToSRV-A(ToR:CR) were observed. A decrease in viral accumulation was observed in plants inoculated with ToSRV-A(ToR:CR+IRD). These results indicate that differences in the CR, but not the IRD, are responsible for the negative interference of ToRMV on ToSRV. [ABSTRACT FROM AUTHOR]

Vasconcellea cauliflora plants exhibiting chlorotic spots, mottling and mosaic with yellowing on leaves were observed in Pune, India in October 2020. The disease incidence was 16%. The samples were screened for cucumber mosaic virus (CMV), papaya ringspot virus, tobacco mosaic virus and zucchini yellow mosaic virus using ELISA. Sap inoculation on healthy V. cauliflora plants using CMV positive DAS-ELISA samples reproduced field symptoms. The transmission upon mechanical inoculation was 100%. For further confirmation, RT-PCR was carried out using CMV coat protein gene specific primers. Amplicons of expected size (c. 650 bp) were obtained in all symptomatic and sap inoculated samples. No amplification was observed in the asymptomatic samples. BLASTn analysis revealed that the amplicon sequence shares more than 99% identity with CMV isolates and in phylogenetic analysis it clustered with CMV IB isolates. This is the first record of V. cauliflora as a natural host of CMV worldwide. [ABSTRACT FROM AUTHOR]

Summary: The hyperosmolality‐gated calcium permeable channel 4.1 (OSCA4.1) belongs to an evolutionarily conserved small family of mechano‐sensitive channels. OSCA members may represent key players in plant resistance to drought and to pathogen infection but are scarcely studied. After screening for resistance to pepino mosaic virus (PepMV) a collection of 1000 mutagenized tomato families, we identified a mutant showing no symptoms and reduced virus accumulation. Resistance was mapped to chromosome 2 between positions 46 309 531 to 47 044 163, where a missense mutation caused the putative truncation of the OSCA4.1 protein. A CRISPR/Cas9 slosca4.1 mutant was resistant to PepMV, but not to tobacco mosaic virus or potato virus X. Inoculation of mutant and wild type tomato protoplasts showed that resistance was expressed in single cells, suggesting a role for SlOSCA4.1 in early viral function(s); congruently, SlOSCA4.1 re‐localized to structures reminiscent of viral replication complexes. We propose that SlOSCA4.1 contributes to the correct regulation of the Ca2+ homeostasis necessary for optimal PepMV infection. PepMV is a pandemic virus that causes significant losses in tomato crops worldwide. In spite of its importance, no tomato‐resistant varieties have been deployed yet; the mutant identified here has great potential to breed tomato varieties resistant to PepMV. [ABSTRACT FROM AUTHOR]

We demonstrate a simple, low-energy method whereby tomato mosaic virus (ToMV) particles can be used to template the production of nanowires and particles consisting of alloys of gold (Au), platinum (Pt) and palladium (Pd) in various combinations. Selective nanowire growth within the inner channel of the particles was achieved using the polymeric capping agent polyvinylpyrrolidone (PVPK30) and the reducing agent ascorbic acid. The reaction conditions also resulted in the deposition of alloy nanoparticles on the external surface of the rods in addition to the nanowire structures within the internal cavity. The resulting materials were characterized using a variety of electron microscopic and spectroscopic techniques, which revealed both the structural and chemical composition of the alloys within the nanomaterials. [ABSTRACT FROM AUTHOR]

Mixed infection by plant viruses is common in nature, but how a key viral protein of one virus affects the infection by heterologous viruses is not yet fully understood. The helper component proteinase (HCPro) is a widely studied RNA silencing suppressor encoded by viruses of the family Potyviridae. Here, we investigated the defence response of Nicotiana tabacum plants overexpressing HCPro of chilli veinal mottle virus (HCPro‐OX) to tobacco mosaic virus and cucumber mosaic virus infection. We monitored the physiological and molecular changes of HCPro‐OX plants in response to virus infection. The results showed that HCPro‐OX plants under virus infection exhibited higher susceptibility at the early stage but stronger tolerance at the later stage compared to wild‐type plants. The tolerance to heterologous virus infection of HCPro‐OX plants corresponded to a lower level of reactive oxygen species accumulation and higher activities of several antioxidant enzymes. Reverse transcription‐quantitative PCR assays showed that the expression of genes related to salicylic acid (SA) pathways was significantly upregulated, but the expression of genes related to the auxin pathways was downregulated at the late stage of virus infection in HCPro‐OX plants compared to wild‐type plants. By contrast, the situation in the early stage of virus infection was reversed. In addition, pretreatment with SA, the auxin naphthylacetic acid (NAA) and their respective inhibitors 1‐aminobenzotriazole (ABT) and naphthalam (NPA) further confirmed the antagonistic effects of SA and NAA in the response of HCPro‐OX plants to heterologous virus infection. Thus, our results demonstrate that HCPro affects heterologous virus infection through SA and auxin pathways. [ABSTRACT FROM AUTHOR]

Abstract—In a series of experiments aimed at testing the reproducibility of stimulated inelastic scattering spectra in suspensions of tobacco mosaic virus nanoparticles, it is found that "freshly prepared" samples feature identical spectra, including those for different virus concentrations in liquid. Sample preparation was accompanied by control measurements within standard biochemical procedures. Using standard control procedures with these virus suspension samples, no changes were detected. However, further laser experiments revealed that virus suspension storage for three years at a temperature of +4°C led to a noticeable change in the stimulated inelastic light scattering spectrum. Differences in the spectra can be due to substructural changes in the virion, for the detection of which standard methods used in virological studies are inapplicable. It is proposed to use stimulated laser light scattering spectroscopy for diagnosing small changes in virions in the native environment. [ABSTRACT FROM AUTHOR]

Background: The discovery and identification of targets is of far‐reaching significance for developing novel pesticide candidates and increasing the probability of success. To explore and identify highly effective tobacco mosaic virus (TMV) helicase‐targeted lead structures, a series of novel phosphonate derivatives containing a 1,2,3‐triazole motif were rationally engineered and their antiviral activity was assessed. Results: Bioassay results showed that the optimized B17 exhibited more potent curative activity (EC50 = 271.5 μg mL−1) against TMV in vivo, which was superior to that of commercial Ribavirin (EC50 = 689.3 μg mL−1). B17 presented a stronger binding capacity through binding analysis with helicase, affording a corresponding value of 12.7 μM. Enzyme activity assay showed B17 exhibited excellent inhibitory activity on TMV helicase (39.2% at 300 μM). Furthermore, molecular docking simulations demonstrated that B17 displayed strong hydrogen‐bond interactions (2.1, 2.1, 2.2, and 3.2 Å) with Ala‐33, Gly‐10, Gly‐8, and Glu‐217 of TMV helicase. Encouragingly, transmission electron microscopy analysis revealed that B17 could remarkably disrupt surface morphology and inhibit TMV proliferation. Additionally, these compounds also displayed potential anti‐CMV (cucumber mosaic virus) and antipathogens (Xanthomonas oryzae pv. oryzae and Xanthomonas axonopodis pv. citri) by expanding their applications in agriculture. Conclusion: Current research demonstrated that B17 could be considered as a potential antiviral agent alternative though targeting TMV helicase. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Plant non-specific lipid transfer proteins (nsLTPs) are small, cysteine-rich proteins that play significant roles in biotic and abiotic stress responses; however, the molecular mechanism of their functions against viral infections remains unclear. In this study, we employed virus-induced gene-silencing and transgenic overexpression to functionally analyse a type-I nsLTP in Nicotiana benthamiana , NbLTP1 , in the immunity response against tobacco mosaic virus (TMV). NbLTP1 was inducible by TMV infection, and its silencing increased TMV-induced oxidative damage and the production of reactive oxygen species (ROS), compromised local and systemic resistance to TMV, and inactivated the biosynthesis of salicylic acid (SA) and its downstream signaling pathway. The effects of NbLTP1 -silencing were partially restored by application of exogenous SA. Overexpressing NbLTP1 activated genes related to ROS scavenging to increase cell membrane stability and maintain redox homeostasis, confirming that an early ROS burst followed by ROS suppression at the later phases of pathogenesis is essential for resistance to TMV infection. The cell-wall localization of NbLTP1 was beneficial to viral resistance. Overall, our results showed that NbLTP1 positively regulates plant immunity against viral infection through up-regulating SA biosynthesis and its downstream signaling component, NONEXPRESSOR OF PATHOGENESIS-RELATED 1 (NPR1), which in turn activates pathogenesis-related genes, and by suppressing ROS accumulation at the later phases of viral pathogenesis. [ABSTRACT FROM AUTHOR]

Grafting is a common horticultural practice used to confer desirable traits between rootstock and scion, including disease resistance. To investigate graft‐conferred resistance against viral diseases a novel heterografting system was developed using Nicotiana benthamiana scions grafted onto different tomato rootstocks. N. benthamiana is normally highly susceptible to tobacco mosaic virus (TMV) infection. However, specific tomato rootstock varieties were found to confer a range of resistance levels to N. benthamiana scions inoculated with TMV. Conferred resistance was associated with delays in virus accumulation and the reduction in virus spread. RNA sequencing analysis showed the enrichment of transcripts associated with disease resistance and plant stress in N. benthamiana scions grafted onto resistance‐inducing tomato rootstocks. Genome sequencing of resistance‐ and nonresistance‐conferring rootstocks was used to identify mobile tomato transcripts within N. benthamiana scions. Within resistance‐induced N. benthamiana scions, enriched mobile tomato transcripts were predominantly associated with defence, stress, and abscisic acid signalling when compared to similar scions grafted onto nonresistance‐inducing rootstock. Combining these findings suggests that graft‐induced resistance is modulated by rootstock scion transcriptional responses and rootstock‐specific mobile transcripts. [ABSTRACT FROM AUTHOR]

Host ranges of plant viruses are poorly known, as studies have focused on pathogenic viruses in crops and adjacent wild plants. High-throughput sequencing (HTS) avoids the bias toward plant–virus interactions that result in disease. Here we study the host ranges of tobamoviruses, important pathogens of crops, using HTS analyses of an extensive sample of plant communities in four habitats of a heterogeneous ecosystem. Sequences of 17 virus operational taxonomic units (OTUs) matched references in the Tobamovirus genus, eight had narrow host ranges, and five had wide host ranges. Regardless of host range, the OTU hosts belonged to taxonomically distant families, suggesting no phylogenetic constraints in host use associated with virus adaptation, and that tobamoviruses may be host generalists. The OTUs identified as tobacco mild green mosaic virus (TMGMV), tobacco mosaic virus (TMV), pepper mild mottle virus, and Youcai mosaic virus had the largest realized host ranges that occurred across habitats and exhibited host use unrelated to the degree of human intervention. This result is at odds with assumptions that contact-transmitted viruses would be more abundant in crops than in wild plant communities and could be explained by effective seed-, contact-, or pollinator-mediated transmission or by survival in the soil. TMGMV and TMV had low genetic diversity that was not structured according to habitat or host plant taxonomy, which indicated that phenotypic plasticity allows virus genotypes to infect new hosts with no need for adaptive evolution. Our results underscore the relevance of ecological factors in host range evolution, in addition to the more often studied genetic factors. [ABSTRACT FROM AUTHOR]

Objective. This research aimed at laboratory simulation of anaerobic digestion of plant biomass contaminated by a mixture of viruses with a wide host range to evaluate if viruses can retain their infectivity and pose a biohazard during the use of plant-based reusable resources such as digestate. Methods. Anaerobic digestion was simulated in air-tight desiccators using anaerobic bags and indicator strips to ensure proper conditions, and using fresh biomass of naturallyvirus-infected plants. In one of the variants, a commercial mixture of anaerobic bacteria and enzymes was also added to see if it would be more efficient in terms of plant digestion and virus inactivation. For serological detection of the tomato mosaic virus, cucumber mosaic virus, and potato virus Y, DAS-ELISA was used with commercial diagnostic kits and positive controls. Absorbance values were measured at 405 nm. Total RNA was extracted using a RNeasy Plant Mini kit following the manufacturer's instructions. Molecular detection of viruses was carried out by two-step RT-PCR using specific oligonucleotide primers available from the literature. To check if the viruses can retain their infectivity after the anaerobic digestion of plant biomass waste, a bioassay was used where Nicotiana rustica plants producing necrotic lesions upon infection with many viruses were mechanically inoculated at the stage of 2 true leaves. For statistical processing of data, the mean and standard error were calculated using Microsoft Excel soft ware. Results. After prolonged anaerobic digestion, tomato mosaic virus was still detected in plant residues, but it is important to note that it was also found in high concentration in the liquid phase. Hence, during the anaerobic digestion, a significant proportion of the virus was successfully eluted from plant residues into the digestate. Interestingly and contrary to tomato mosaic virus, cucumber mosaic virus has not practically eluted to the liquid phase. On the other hand, potato virus Y was supposedly completely decomposed during the anaerobic digestion of plant biomass waste, as it was not detected either by ELISA or by PCR. It is likely that not all of the investigated viruses represent a potential biohazard in the context of the use of renewable resources. RT-PCR outcomes showed the presence of tomato mosaic virus and cucumber mosaic virus both in the solid and the liquid phases, augmenting the ELISA data. Lastly, bioassay results have confirmed that at least tomato mosaic tobamovirus may escape the aggressive process of waste decomposition and remain able to infect host plants raising issues about the safety of reusable plant resources. Conclusions. The survival of plant viruses during plant biomass waste processing was studied in the context of biological safety. The preservation of serological markers and genetic signatures of model viruses in the process of anaerobic digestion has been confirmed. For the first time, it was proven that tomato mosaic virus may preserve its infectivity during long-term incubation of plant biomass waste in anaerobic conditions, which requires further attention when using renewable resources. On the example of PVY, it has been shown that potyviruses do not survive during anaerobic digestion and therefore may not pose an epidemic risk in using processed renewable resources. It has been demonstrated that artificial introduction of anaerobic microorganisms does not have a decisive effect on the stability of the studied model viruses. [ABSTRACT FROM AUTHOR]

In this study, we used RT-qPCR to examine how PR genes were expressed in model tomato (Solanum lycopersicum L.) plants that had been infected with TMV or CMV. Under greenhouse conditions, the indirect ELISA data showed that both viruses were detected for the first time at 6 dpi. Then, the levels of accumulation increased very quickly, reaching a peak of 15 dpi. During the course of the study (1–15 dpi), the Delta CT, NormFinder, BestKeeper, and GeNorm software tools revealed that the β-actin gene was the most informative reference gene in the virally infected tomato tissues. For both the TMV- and CMV-infected tomato plants, the transcriptional expression levels of most tested genes changed between activation and repression, especially in the first 12 dpi. Compared to mock-inoculated plants, the expression levels of PR-1 were induced at all time intervals except at 8 dpi for CMV and at 6, 7, and 8 dpi for TMV infection. Conversely, the greater activation and accumulation of both viruses were associated with the greater up-regulation of PR-2 at 8 dpi, with relative expression levels of 7.28- and 5.84-fold for TMV and CMV, respectively. The up-regulated expression of PR-3, PR-4, and PR-7 was shown at 4 dpi. In contrast, the PR-5 gene was inhibited in TMV at 1 dpi until 9 dpi, and the induction of this gene at 10 dpi increased by 1.72-fold, but PR-5 was observed to up-regulate the expression of CMV at 1 dpi. This study provides the first valuable information on the comparative transcriptional levels of these tomato genes between TMV and CMV infections.

The article "In the Spotlight: The plant growth‐defence dilemma: a hormonal balancing act" published in Physiologia Plantarum discusses how plants face challenges in balancing growth and defense mechanisms, particularly in the context of increasing temperatures and new diseases due to climate change. Research by Huang et al. highlights the roles of plant hormones, such as salicylic acid and auxins, in this balancing act, showing how they interact with catalases to either boost defenses or enhance growth. Understanding these interactions is crucial for plant health and food security, especially as climate change impacts the prevalence of plant viruses like TMV and CMV, emphasizing the need for targeted genetic engineering strategies. [Extracted from the article]

Essential in halting the COVID-19 pandemic caused by SARS-CoV-2, it is crucial to have stable, effective, and easy-to-manufacture vaccines. We developed a potential vaccine using a tobacco mosaic virus (TMV) epitope display model presenting peptides derived from the SARS-CoV-2 spike protein. The TMV-epitope fusions in laboratory tests demonstrated binding to the SARS-CoV-2 polyclonal antibodies. The fusion constructs maintained critical epitopes of the SARS-CoV-2 spike protein, and two in particular spanned regions of the receptor-binding domain that have mutated in the more recent SARS-CoV-2 variants. This would allow for the rapid modification of vaccines in response to changes in circulating variants. The TMV-peptide fusion constructs also remained stable for over 28 days when stored at temperatures between −20 and 37 °C, an ideal property when targeting developing countries. Immunogenicity studies conducted on BALB/c mice elicited robust antibody responses against SARS-CoV-2. A strong IFNγ response was also observed in immunized mice. Three of the six TMV-peptide fusion constructs produced virus-neutralizing titers, as measured with a pseudovirus neutralization assay. These TMV-peptide fusion constructs can be combined to make a multivalent vaccine that could be adapted to meet changing virus variants. These findings demonstrate the development of a stable COVID-19 vaccine candidate by combining SARS-CoV-2 spike protein-derived peptides presented on the surface of a TMV nanoparticle.

Rotavirus infection is a leading cause of severe dehydrating gastroenteritis in children under 5 years of age. Although rotavirus-associated mortality has decreased considerably because of the introduction of the worldwide rotavirus vaccination, the global burden of rotavirus-associated gastroenteritis remains high. Current vaccines have a number of disadvantages; therefore, there is a need for innovative approaches in rotavirus vaccine development. In the current study, a universal recombinant rotavirus antigen (URRA) for a novel recombinant vaccine candidate against rotavirus A was obtained and characterised. This antigen included sequences of the VP8* subunit of rotavirus spike protein VP4. For the URRA, for the first time, two approaches were implemented simultaneously—the application of a highly conserved neutralising epitope and the use of the consensus of the extended protein’s fragment. The recognition of URRA by antisera to patient-derived field rotavirus isolates was proven. Plant virus-based spherical particles (SPs), a novel, effective and safe adjuvant, considerably enhanced the immunogenicity of the URRA in a mouse model. Given these facts, a URRA + SPs vaccine candidate is regarded as a prospective basis for a universal vaccine against rotavirus.

Reversibly glycosylated polypeptides (RGPs) have been identified in many plant species and play an important role in cell wall formation, intercellular transport regulation, and plant–virus interactions. Most plants have several RGP genes with different expression patterns depending on the organ and developmental stage. Here, we report on four members of the RGP family in N. benthamiana. Based on a homology search, NbRGP1-3 and NbRGP5 were assigned to the class 1 and class 2 RGPs, respectively. We demonstrated that NbRGP1–3 and 5 mRNA accumulation increases significantly in response to tobacco mosaic virus (TMV) infection. Moreover, all identified class 1 NbRGPs (as distinct from NbRGP5) suppress TMV intercellular transport and replication in N. benthamiana. Elevated expression of NbRGP1–2 led to the stimulation of callose deposition at plasmodesmata, indicating that RGP-mediated TMV local spread could be affected via a callose-dependent mechanism. It was also demonstrated that NbRGP1 interacts with TMV movement protein (MP) in vitro and in vivo. Therefore, class 1 NbRGP1–2 play an antiviral role by impeding intercellular transport of the virus by affecting plasmodesmata callose and directly interacting with TMV MP, resulting in the reduced viral spread and replication. [ABSTRACT FROM AUTHOR]

Catalases (CATs) play important roles in plant growth, development and defense responses. Previous studies have shown that CATs exhibit different or even opposite effects on plant immunity in different plant–pathogen interactions, but little is known about the mechanisms. In this study, Nicotiana tabacum plants with overexpression or knockout of CAT genes, tobacco mosaic virus (TMV) and cucumber mosaic virus (CMV) were employed to investigate the role of CAT in compatible plant–virus interactions. The results showed that there were dynamic changes in the effect of CAT on N. tabacum defense responses. Overexpression of catalase 1 (CAT1) and catalase 3 (CAT3) improved N. tabacum resistance in the early stage of virus infection but depressed it during the late stages of pathogenesis, especially in CAT3 overexpressing plants. The lower level of electrolyte leakage, lower contents of malonaldehyde and hydrogen peroxide (H2O2), higher activities of antioxidant enzymes and improved functions of photosystem II corresponded to the milder symptoms and higher resistance of infected tobacco plants. In addition, the infection of TMV and CMV resulted in expression changes of CATs in tobacco plants, and pretreatment with H2O2 facilitated TMV and CMV infection. Further experiments showed that the content of salicylic acid (SA) and the expression of genes related to SA signaling pathway were positively correlated with plant resistance, whereas auxin and its related signaling pathway were related to the viral susceptibility of plants. Taken together, our results demonstrated that CAT1 and CAT3 mediated tobacco resistance to virus infection through crosstalk between SA and auxin signaling pathways. [ABSTRACT FROM AUTHOR]

The tomato Tm‐22 gene was considered to be one of the most durable resistance genes in agriculture, protecting against viruses of the Tobamovirus genus, such as tomato mosaic virus (ToMV) and tobacco mosaic virus (TMV). However, an emerging tobamovirus, tomato brown rugose fruit virus (ToBRFV), has overcome Tm‐22, damaging tomato production worldwide. Tm‐22 encodes a nucleotide‐binding leucine‐rich repeat (NLR) class immune receptor that recognizes its effector, the tobamovirus movement protein (MP). Previously, we found that ToBRFV MP (MPToBRFV) enabled the virus to overcome Tm‐22‐mediated resistance. Yet, it was unknown how Tm‐22 remained durable against other tobamoviruses, such as TMV and ToMV, for over 60 years. Here, we show that a conserved cysteine (C68) in the MP of TMV (MPTMV) plays a dual role in Tm‐22 activation and viral movement. Substitution of MPToBRFV amino acid H67 with the corresponding amino acid in MPTMV (C68) activated Tm‐22‐mediated resistance. However, replacement of C68 in TMV and ToMV disabled the infectivity of both viruses. Phylogenetic and structural prediction analysis revealed that C68 is conserved among all Solanaceae‐infecting tobamoviruses except ToBRFV and localizes to a predicted jelly‐roll fold common to various MPs. Cell‐to‐cell and subcellular movement analysis showed that C68 is required for the movement of TMV by regulating the MP interaction with the endoplasmic reticulum and targeting it to plasmodesmata. The dual role of C68 in viral movement and Tm‐22 immune activation could explain how TMV was unable to overcome this resistance for such a long period. [ABSTRACT FROM AUTHOR]

Tomato mottle mosaic virus (ToMMV) is an emerging seed-transmissible tobamovirus that infects tomato and pepper. Since the first report in 2013 in Mexico, ToMMV has spread worldwide, posing a serious threat to the production of both crops. To prevent the spread of this virus, early and accurate detection of infection is required. In this study, we developed a detection method for ToMMV based on reverse-transcription loop-mediated isothermal amplification (RT-LAMP). A LAMP primer set was designed to target the genomic region spanning the movement protein and coat protein genes, which is a highly conserved sequence unique to ToMMV. This RT-LAMP detection method achieved 10-fold higher sensitivity than conventional RT-polymerase chain reaction methods and obtained high specificity without false positives for closely related tobamoviruses or healthy tomato plants. This method can detect ToMMV within 30 min of direct sampling of an infected tomato leaf using a toothpick and therefore does not require RNA purification. Given its high sensitivity, specificity, simplicity, and rapidity, the RT-LAMP method developed in this study is expected to be valuable for point-of-care testing in field surveys and for large-scale testing. [ABSTRACT FROM AUTHOR]

Viral infection, which entails synthesis of viral proteins and active reproduction of the viral genome, effects significant changes in the functions of many intracellular systems in plants. Along with these processes, a virus has to suppress cellular defense to create favorable conditions for its successful systemic spread in a plant. The virus exploits various cellular factors of a permissive host modulating its metabolism as well as local and systemic transport of macromolecules and photoassimilates. The Nicotiana benthamiana stress-induced gene encoding Kunitz peptidase inhibitor-like protein (KPILP) has recently been shown to be involved in chloroplast retrograde signaling regulation and stimulation of intercellular transport of macromolecules. In this paper we demonstrate the key role of KPILP in the development of tobamovius infection. Systemic infection of N. benthamiana plants with tobacco mosaic virus (TMV) or the closely related crucifer-infecting tobamovirus (crTMV) induces a drastic increase in KPILP mRNA accumulation. KPILP knockdown significantly reduces the efficiency of TMV and crTMV intercellular transport and reproduction. Plants with KPILP silencing become partially resistant to tobamovirus infection. Therefore, KPILP could be regarded as a novel proviral factor in the development of TMV and crTMV infection in N. benthamiana plants. [ABSTRACT FROM AUTHOR]

Vaccines are the cornerstone of infectious disease control and prevention. The outbreak of SARS-CoV-2 has confirmed the urgent need for a new approach to the design of novel vaccines. Plant viruses and their derivatives are being used increasingly for the development of new medical and biotechnological applications, and this is reflected in a number of preclinical and clinical studies. Plant viruses have a unique combination of features (biosafety, low reactogenicity, inexpensiveness and ease of production, etc.), which determine their potential. This review presents the latest data on the use of plant viruses with different types of symmetry as vaccine components and adjuvants in cancer immunotherapy. The discussion concludes that the most promising approaches might be those that use structurally modified plant viruses (spherical particles) obtained from the Tobacco mosaic virus. These particles combine high adsorption properties (as a carrier) with strong immunogenicity, as has been confirmed using various antigens in animal models. According to current research, it is evident that plant viruses have great potential for application in the development of vaccines and in cancer immunotherapy. [ABSTRACT FROM AUTHOR]

Agricultural production without pollution is possible using bioactive plant compounds, which include essential oils as important substances of plant origin. The aim of this study was to evaluate the antiphytoviral and antibacterial potentials of lipid (essential oil, EO) and water (hydrosol, HY) extracts from five Veronica species (Plantaginaceae) obtained by Clevenger hydrodistillation (HD) and microwave-assisted extraction (MAE), with analysis by gas chromatography coupled with mass spectrometry. The antiphytoviral activities of both extracts were tested on local host plants infected with tobacco mosaic virus (TMV). The antibacterial potential was tested against ten strains of opportunistic pathogens using the broth microdilution test. Species V. chamaedrys EO-MAE extract, V. arvensis EO from both extractions and V. montana, V. serpyllifolia, and V. persica EO-HD extracts were more effective in inhibiting TMV infection. Furthermore, HY- HD extracts of V. arvensis, V. chamaedrys and V. persica showed significant antiphytoviral activity. HY fractions had no effect on bacterial growth, regardless of the Veronica species tested, likely due to the fact that the maximum concentrations of the HY fractions tested in this study were low (1.83 and 2.91 mg/mL). EOs showed significant antibacterial activity independent of the extraction method. Notably, V. chamaedrys EO-MAE fraction, showed significantly better activity against Listeria monocytogenes and Enterococcus faecalis. Also, the EO-HD fraction of V. arvensis showed slightly better antibacterial activity. By combining extracts and using different extraction methods, valuable bioproducts can be obtained from the investigated Veronica species for safe use in agricultural production and food conservation. [ABSTRACT FROM AUTHOR]

Plants respond to heat stress by producing heat-shock proteins. These are regulated by heat-shock promoters containing regulatory elements, which can be harnessed to control protein expression both temporally and spatially. In this study, we designed heat-inducible promoters to produce the diterpene casbene in Nicotiana benthamiana, through a multi-step metabolic pathway. To potentially increase gene transcription, we coupled heat-shock elements from Arabidopsis thaliana Hsp101 or Glycine max GmHsp17.3-B promoters, CAAT and TATA boxes from CaMV 35S, and the 5′UTR from the tobacco mosaic virus. The resulting four chimeric promoters fused to a green fluorescent protein (GFP) reporter showed that the variant Ara2 had the strongest fluorescent signal after heat shock. We next created a 4-gene cassette driven by the Ara2 promoter to allow for exogenous synthesis of casbene and transformed this multigene construct along with a selectable marker gene into Nicotiana benthamiana. Metabolic analysis on the transgenic lines revealed that continuous heat outperforms heat shock, with up to 1 μg/mg DW of casbene detected after 32 h of uninterrupted 40 °C heat. These results demonstrate the potential of heat-inducible promoters as synthetic biology tools for metabolite production in plants. [ABSTRACT FROM AUTHOR]

Simple Summary: The Epstein–Barr virus is a human herpesvirus that infects the majority of the human population. Initially, infection can cause mild symptoms in infants and children, but can also cause short-lived infectious mononucleosis in teenage and young adults. But, in a small minority of older patients, persistent expression of virus antigens can cause B cell transformation to a cancer. Patients with Epstein–Barr virus-positive B cell lymphoma are often more resistant to treatment with standard chemotherapy. One difficulty in developing new methods of treatment is that Epstein–Barr only infects human cells, and there are no mouse B cell models of the disease. Our goal was to develop a mouse model of disease, in order to study the potential for effective vaccination against an Epstein–Barr virus antigen. We were successful in generating a mouse B cell line expressing latent membrane protein 1, and could show that vaccination protected mice against B cell tumors that expressed latent membrane protein 1. This system will allow us to further study the potential for vaccination in patients with cancer, to help the immune system recognize and eliminate tumor cells. In this study, we demonstrate that expression of viral latent membrane protein 1 (LMP1) in a mouse B cell line renders the animals responsive to protection from a 38C13-LMP1 tumor challenge with a novel vaccine. The Epstein–Barr virus (EBV) preferentially infects circulating B lymphocytes, has oncogenic potential, and is associated with a wide variety of B cell lymphomas. EBV is ectotrophic to human cells, and currently there are no B cell animal models of EBV-associated lymphoma that can be used to investigate vaccine immunotherapy. Since most EBV-infected human tumor cells express latent membrane protein 1 (LMP1) on their surface, this viral antigen was tested as a potential target for an anticancer vaccine in a mouse model. Here, we describe a new mouse model of LMP1-expressing B cell lymphoma produced with plasmid transduction of 38C13 into mouse B cells. The expression of LMP-1 was confirmed with a western blot analysis and immunocytochemistry. We then designed a novel LMP1 vaccine, by fusing viral antigen LMP1 surface loop epitopes to the surface of a viral antigen carrier, the Tobacco Mosaic virus (TMV). Vaccinated mice produced high titer antibodies against the TMV-LMP1 vaccine; however, cellular responses were at the baseline, as measured with IFNγ ELISpot. Despite this, the vaccine showed significant protection from a 38C13-LMP1 tumor challenge. To provide additional immune targets, we compared TMV-LMP1 peptide immunization with DNA immunization with the full-length LMP1 gene. Anti-LMP1 antibodies were significantly higher in TMV-LMP1-vaccinated mice compared to the DNA-immunized mice, but, as predicted, DNA-vaccinated mice had improved cellular responses using IFNγ ELISpot. Surprisingly, the TMV-LMP1 vaccine provided protection from a 38C13-LMP1 tumor challenge, while the DNA vaccine did not. Thus, we demonstrated that LMP1 expression in a mouse B cell line is responsive to antibody immunotherapy that may be applied to EBV-associated disease. [ABSTRACT FROM AUTHOR]

RNA viruses tend to mutate during transmission and host infection, which is critical to viral adaptation and evolution. Tomato mosaic virus (ToMV) is a member of the genus Tobamovirus (family Virgaviridae) and an economically important virus with detrimental effects on tomatoes worldwide. Although the ToMV gene sequences have been completed in China, their genetic diversity and population structure remain unclear. We collected 425 tomato samples from tomato-growing areas in three northern Chinese provinces 2016. Reverse transcription PCR results showed that the average incidence of the virus in the field samples was 67.15%, and ToMV was detected in all test areas. The analysis of ToMV single nucleotide polymorphisms in China showed that ToMV was evolutionarily conserved, and the variation in the whole genome was uneven. Pairwise identity analysis showed significant variability in genome sequences among ToMV strains with genomic nucleotide identities of 73.2–99.6%. The ToMV population in the northern Chinese provinces had purification and selection functions, which were beneficial in the evolution of the ToMV population. Although there has been some distribution of ToMV strains in China, the virus was generally stabilized as a uniform strain under the pressure of purification selection. Our findings show how to monitor the prevalent strains of ToMV and their virulence in China and provide useful information for its prevention and control. [ABSTRACT FROM AUTHOR]

pepper plants treated with the same Tobacco mosaic virus (TMV) showed no signs and symptoms of disease, in contrast to plants treated with these compounds as a control, which was thought to act as an inhibitory start to the viral infection. This resistance was sparked by the pepper plants' exposure to the TMV virus. Furthermore, the ELISA test findings indicated a wave reaction based on the yellow color that developed in the ELISA plate wells containing the extracts of pepper plants that have been treated with Ganoderma lucidum extract. When compared to the control treatment, which is infected with the virus alone, the wave absorption amount at 405 nm was 0.296 nm, indicating that the highest percentage of virus duplication occurred (69%) after the second week of the G. lucidum treatment and before six days of infection. On the other hand, 58% of the virus was duplicated when the G. lucidum extract was sprayed three days prior to the virus infection, and the wave absorption value was 0.404 nm. The values of wave absorption following splashing with the same extract were 0.612 and 0.598 after three and six days of virus infection, respectively, and the percentages of virus duplication were 37 and 38%. Lastly, it should be emphasized that, rather than using chemical pesticides that are unsafe to use against the causes of plant diseases, more research must be done in the future regarding the extraction of these anti-viral plant extracts as well as extracts of naturally occurring substances that are environmentally friendly. [ABSTRACT FROM AUTHOR]

Botrytis cinerea is the causative agent of gray mold disease, and infects more than 1400 plant species, including important crop plants. In tomato, B. cinerea causes severe damage in greenhouses and post-harvest storage and transport. Plant viruses of the Tobamovirus genus cause significant damage to various crop species. In recent years, the tobamovirus tomato brown rugose fruit virus (ToBRFV) has significantly affected the global tomato industry. Most studies of plant-microbe interactions focus on the interaction between the plant host and a single pathogen, however, in agricultural or natural environments, plants are routinely exposed to multiple pathogens. Here, we examined how preceding tobamovirus infection affects the response of tomato to subsequent infection by B. cinerea. We found that infection with the tobamoviruses tomato mosaic virus (ToMV) or ToBRFV resulted in increased susceptibility to B. cinerea. Analysis of the immune response of tobamovirus-infected plants revealed hyperaccumulation of endogenous salicylic acid (SA), upregulation of SA-responsive transcripts, and activation of SA-mediated immunity. Deficiency in SA biosynthesis decreased tobamovirus-mediated susceptibility to B. cinerea, while exogenous application of SA enhanced B. cinerea symptoms. These results suggest that tobamovirus-mediated accumulation of SA increases the plants' susceptibility to B. cinerea, and provide evidence for a new risk caused by tobamovirus infection in agriculture. [ABSTRACT FROM AUTHOR]

Numerous, diverse plant viruses encode movement proteins (MPs) that aid the virus movement through plasmodesmata, the plant intercellular channels. MPs are essential for virus spread and propagation in distal tissues, and several unrelated MPs have been identified. The 30K superfamily of MPs (named after the molecular mass of tobacco mosaic virus (TMV) MP, the classical model of plant virology) is the largest and most diverse MP variety, represented in 16 virus families, but its evolutionary origin remained obscure. Here, we show that the core structural domain of the 30K MPs is homologous to the jelly-roll domain of the capsid proteins (CPs) of small RNA and DNA viruses, in particular, those infecting plants. The closest similarity was observed between the 30K MPs and the CPs of the viruses in the families Bromoviridae and Geminiviridae. We hypothesize that the MPs evolved via duplication or horizontal acquisition of the CP gene in a virus that infected an ancestor of vascular plants, followed by neofunctionalization of one of the paralogous CPs, potentially through the acquisition of unique N- and C-terminal regions. During the subsequent coevolution of viruses with diversifying vascular plants, the 30K MP genes underwent explosive horizontal spread among emergent RNA and DNA viruses, likely permitting viruses of insects and fungi that coinfected plants to expand their host ranges, molding the contemporary plant virome. Movement proteins, a signature of plant RNA and DNA viruses, enable cell-to-cell movement of the virus via plasmodesmata channels. This study shows that the most common movement proteins of the 30K superfamily originated from jelly-roll capsid proteins of plant viruses, emphasizing the key role of exaptation in virus evolution. [ABSTRACT FROM AUTHOR]

The first infectious agent to bear the name 'virus' was described in 1898: a plant pathogen called tobacco mosaic virus that infects a wide range of plants and results in a yellow mosaic of the leaves. Since then, the study of plant viruses has facilitated new discoveries in both virology and plant biology. Traditionally, research has focused on viruses that cause severe disease in plants used for human and animal food or recreation. However, closer inspection of the plant-associated virome is now revealing interactions that range from pathogenic to symbiotic. Although they are often studied in isolation, plant viruses are usually found as part of a broader community that includes other plant-associated microbes and pests. For example, biological vectors of plant viruses (arthropods, nematodes, fungi, and protists) can facilitate the transmission of viruses between plants in an intricate interaction. To enhance transmission, viruses can induce the plant to 'lure' the vector by modulating plant chemistry and defenses. Once delivered to a new host, viruses are dependent on specific proteins that modify the structural components of the cell to enable transport of viral proteins and genomic material. Links between antiviral plant defenses and key steps in virus movement and transmission are being revealed. Upon infection, a suite of antiviral responses is triggered, including the expression of resistance genes — a favored strategy to control plant viruses. In this primer, we discuss these features and more, highlighting the exciting world of plant–virus interactions. In this Primer, Cesar Xavier and Anna Whitfield introduce plant viruses, and discuss the many ways in which the interactions between virus and plant hosts influence plant biology. [ABSTRACT FROM AUTHOR]

Strain K-11 was isolated from the highly saline brown semi-desert soil of the Astrakhan region. Based on analysis of the 16S rRNA gene sequence, this strain was identified as Streptomyces carpaticus K-11 RCAM04697 (SCPM-O-B-9993). Whole genome sequencing of the strain was performed. Phytotoxicity, antiviral, antioxidant, antifungal, and insecticidal activities of the strain were studied. All extracts and suspensions of S. carpaticus strain RCAM04697 had plant-stimulating activity. Antiviral properties was exhibited as suppression of development and propagation of viral pathogens in laboratory conditions: Tomato mosaic virus (ToMV)—26.3%, Cucumber mosaic virus (CMV)—33.8%, Y-Potato virus (Potato Y potyvirus, PVY)—51.3%, Potato X-virus (PVX) (Potato X potyvirus, PVX)—41.3%. The highest antioxidant activity was shown by a suspension of S. carpaticus strain RCAM04697 (88.8%) and its aqueous-alcoholic (20 : 80) extract (76.0%). The strain inhibited growth of the phytopathogenic fungus Fusarium sporotrichioides to varying degrees. The insecticidal activity against Aphis fabae after 6 h of treatment was 100% in the variants with suspension treatment, water-alcohol (80 : 20, 50 : 50), methanol, and hexane extracts. The metabolites of the S. carpaticus RCAM04697 strain included flavonoids, alkaloids, glycosides, organic acids (isocitric, acetic, fumaric, lactic, pyruvic, and malic), alcohols, aldehydes, hydrocarbons, ethers, sulfur-containing compounds, and other groups of low-molecular weight organic compounds. [ABSTRACT FROM AUTHOR]

In this study, we used RT-qPCR to examine how PR genes were expressed in model tomato (Solanum lycopersicum L.) plants that had been infected with TMV or CMV. Under greenhouse conditions, the indirect ELISA data showed that both viruses were detected for the first time at 6 dpi. Then, the levels of accumulation increased very quickly, reaching a peak of 15 dpi. During the course of the study (1–15 dpi), the Delta CT, NormFinder, BestKeeper, and GeNorm software tools revealed that the β-actin gene was the most informative reference gene in the virally infected tomato tissues. For both the TMV- and CMV-infected tomato plants, the transcriptional expression levels of most tested genes changed between activation and repression, especially in the first 12 dpi. Compared to mock-inoculated plants, the expression levels of PR-1 were induced at all time intervals except at 8 dpi for CMV and at 6, 7, and 8 dpi for TMV infection. Conversely, the greater activation and accumulation of both viruses were associated with the greater up-regulation of PR-2 at 8 dpi, with relative expression levels of 7.28- and 5.84-fold for TMV and CMV, respectively. The up-regulated expression of PR-3, PR-4, and PR-7 was shown at 4 dpi. In contrast, the PR-5 gene was inhibited in TMV at 1 dpi until 9 dpi, and the induction of this gene at 10 dpi increased by 1.72-fold, but PR-5 was observed to up-regulate the expression of CMV at 1 dpi. This study provides the first valuable information on the comparative transcriptional levels of these tomato genes between TMV and CMV infections. [ABSTRACT FROM AUTHOR]

Tobacco mosaic virus (TMV) is a major pathogen affecting tomato plants worldwide. The efficacy of silver nanoparticles (Ag-NPs) mediated by Punica granatum biowaste peel extract in mitigating the negative impact of TMV infection on tomato growth and oxidative stress was investigated through scanning electron microscopy (SEM), transmission electron microscopy (TEM), UV-Visible (UV-Vis) spectrophotometer, X-ray Diffraction (XRD), dynamic light scattering (DLS), zeta potential, energy-dispersive X-ray spectroscopy (EDX), and Fourier-transform infrared spectra (FTIR). Results of SEM analysis of green Ag-NPs revealed the presence of condensed spherical or round NPs with diameters ranging between 61 and 97 nm. TEM confirmed the SEM results and showed round-shaped Ag-NPs with an average size of 33.37 ± 12.7 nm. The elemental analysis (EDX) of prepared Ag-NPs revealed the presence of elemental Ag as a major peak (64.43%) at 3–3.5 KeV. The FTIR revealed several functional groups on the prepared Ag-NPs, for which three treatment strategies for Ag-NP applications were evaluated in the greenhouse study and compared to inoculated TMV and control plants: pre-infection treatment (TB), post-infection treatment (TA), and dual treatment (TD). The results showed that the TD strategy is the most effective in improving tomato growth and reducing viral replication, whereas all Ag-NP treatments (TB, TA, and TD) were found to significantly increase expression of the pathogenesis-related (PR) genes PR-1 and PR-2, as well as polyphenolic compounds, HQT, and C4H genes compared to control plants. In contrast, the flavonoid content of tomato plants was not affected by the viral infection, while the phenolic content was significantly reduced in the TMV group. Furthermore, TMV infection led to a significant increase in oxidative stress markers MDA and H2O2, as well as a reduction in the enzymatic activity of the antioxidants PPO, SOD, and POX. Our results clearly showed that the application of Ag-NPs on TMV-infected plants reduces virus accumulation, delays viral replication in all treatments, and greatly enhances the expression of the CHS gene involved in flavonoid biosynthesis. Overall, these findings suggest that treatment with Ag-NPs may be an effective strategy to mitigate the negative impact of TMV infection on tomato plants. [ABSTRACT FROM AUTHOR]

A convenient two-step method is reported for the ligation of alkoxyamine- or hydrazine-bearing cargo to proline N-termini. Using this approach, bifunctional proline N-terminal bioconjugates are constructed and proline N-terminal proteins are immobilized.

Virus infection in chili plants may cause various symptoms. The complexity of the symptoms and human vision ability often become limiting factors during disease investigations. Digital image analysis is expected to become a method to assist in comprehensively describing the symptoms of plant viruses. A disease survey was conducted on cayenne pepper fields in Southeast Sulawesi Province to observe symptoms of virus infection virtually and to record the symptomatic plant using an RGB camera. The split-channel method is used to process images, followed by multidimensional scaling statistical analysis. Later on, viruses associated with plants were detected serologically. Single or mixed infection of Tobacco mosaic virus, Cucumber mosaic virus, Chili veinal mottle virus, and Pepper mottle virus was confirmed by plant leaves showing yellow-mosaic and mottle symptoms. The digital image analysis method could show variations in the characteristics of symptoms based on digital numbers in that cannot be recognized based on the observation of visual symptoms. A new approach to study the interactions between plant infecting viruses and their effects based on image analysis has also been developed during this research. This method needs to be further validated through testing under controlled conditions, such as inoculating plants with a predetermined type of virus.

Compounds with acylhydrazone fragments contain amide and imine groups that can act as electron donors and acceptors, so they are easier to bind to biological targets and thus generally exhibit significant biological activity. In this work, acylhydrazone fragments were introduced to the C-14 or C-11 position of matrine, a natural alkaloid, aiming to enhance their biological activities. The result of this bioassay showed that many synthesized compounds exhibited excellent anti-virus activity against the tobacco mosaic virus (TMV). Seventeen out of 25 14-acylhydrazone matrine derivatives and 17 out of 20 11-butanehydrazone matrine derivatives had a higher inhibitory activity against TMV than the commercial antiviral agent Ribavirin (the in vitro activity, in vivo inactivation, curative and protection activities at 500 µg/mL were 40.9, 36.5 ± 0.9, 38.0 ± 1.6 and 35.1 ± 2.2%, respectively), and four 11-butanehydrazone matrine derivatives even had similar to or higher activity than the most efficient antiviral agent Ningnanmycin (55.4, 57.8 ± 1.4, 55.3 ± 0.5 and 60.3 ± 1.2% at 500 µg/mL for the above four test modes). Among them, the N-benzyl-11-butanehydrazone of matrine formed with 4-bromoindole-3-carboxaldehyde exhibited the best anti-TMV activity (65.8, 71.8 ± 2.8, 66.8 ± 1.3 and 69.5 ± 3.1% at 500 µg/mL; 29, 33.5 ± 0.7, 24.1 ± 0.2 and 30.3 ± 0.6% at 100 µg/mL for the above four test modes), deserving further investigation as an antiviral agent. Other than these, the two series of acylhydrazone-containing matrine derivatives were evaluated for their insecticidal and fungicidal activities. Several compounds were found to have good insecticidal activities against diamondback moth (Plutella xylostella) and mosquito larvae (Culex pipiens pallens), showing broad biological activities. [ABSTRACT FROM AUTHOR]

Tobacco mosaic virus (TMV) is a systemic virus that poses a serious threat to crops worldwide. In the present study, a series of novel 1-phenyl-4-(1,3,4-thiadiazole-5-thioether)-1H-pyrazole-5-amine derivatives was designed and synthesized. In vivo antiviral bioassay results indicated that some of these compounds exhibited excellent protective activity against TMV. Among the compounds, E2 (EC50 = 203.5 μg/mL) was superior to the commercial agent ningnanmycin (EC50 = 261.4 μg/mL). Observation of tobacco leaves infected with TMV-GFP revealed that E2 could effectively inhibit the spread of TMV in the host. Further plant tissue morphological observation indicated that E2 could induce the tight arrangement and alignment of the spongy mesophyll and palisade cells while causing stomatal closure to form a defensive barrier to prevent viral infection in the leaves. In addition, the chlorophyll content of tobacco leaves was significantly increased after treatment with E2, and the net photosynthesis (Pn) value was also increased, which demonstrated that the active compound could improve the photosynthetic efficiency of TMV-infected tobacco leaves by maintaining stable chlorophyll content in the leaves, thereby protecting host plants from viral infection. The results of MDA and H2O2 content determination revealed that E2 could effectively reduce the content of peroxides in the infected plants, reducing the damage to the plants caused by oxidation. This work provides an important support for the research and development of antiviral agents in crop protection. [ABSTRACT FROM AUTHOR]

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Plant diseases and insect pests seriously affect the yield and quality of crops and are difficult to control. Natural products are an important source for the discovery of new pesticides. In this work, naphthoquinones plumbagin and juglone were selected as parent structures, and a series of their derivatives were designed, synthesized and evaluated for their fungicidal activities, antiviral activities and insecticidal activities. We found that the naphthoquinones have broad-spectrum anti-fungal activities against 14 types of fungus for the first time. Some of the naphthoquinones showed higher fungicidal activities than pyrimethanil. Compounds I, I-1e and II-1a emerged as new anti-fungal lead compounds with excellent fungicidal activities (EC50 values: 11.35–17.70 µg/mL) against Cercospora, arachidicola Hori. Some compounds also displayed good to excellent antiviral activities against the tobacco mosaic virus (TMV). Compounds I-1f and II-1f showed similar level of anti-TMV activities with ribavirin, and could be used as new antiviral candidates. These compound also exhibited good to excellent insecticidal activities. Compounds II-1d and III-1c displayed a similar level of insecticidal activities with matrine, hexaflumuron and rotenone against Plutella xylostella. In current study, plumbagin and juglone were discovered as parent structures, which lays a foundation for their application in plant protection. [ABSTRACT FROM AUTHOR]

Different diseases are observed in vegetables, fruits, cereals, and commercial crops by farmers and agricultural experts. Nonetheless, this evaluation process is time-consuming, and initial symptoms are primarily visible at microscopic levels, limiting the possibility of an accurate diagnosis. This paper proposes an innovative method for identifying and classifying infected brinjal leaves using Deep Convolutional Neural Networks (DCNN) and Radial Basis Feed Forward Neural Networks (RBFNN). We collected 1100 images of brinjal leaf disease that were caused by five different species (Pseudomonas solanacearum, Cercospora solani, Alternaria melongenea, Pythium aphanidermatum, and Tobacco Mosaic Virus) and 400 images of healthy leaves from India's agricultural form. First, the original plant leaf is preprocessed by a Gaussian filter to reduce the noise and improve the quality of the image through image enhancement. A segmentation method based on expectation and maximization (EM) is then utilized to segment the leaf's-diseased regions. Next, the discrete Shearlet transform is used to extract the main features of the images such as texture, color, and structure, which are then merged to produce vectors. Lastly, DCNN and RBFNN are used to classify brinjal leaves based on their disease types. The DCNN achieved a mean accuracy of 93.30% (with fusion) and 76.70% (without fusion) compared to the RBFNN (82%—without fusion, 87%—with fusion) in classifying leaf diseases. [ABSTRACT FROM AUTHOR]

Cucumber mosaic virus (CMV) is vectored by aphids, including Myzus persicae. Tobacco (Nicotiana tabacum 'Xanthi') plants infected with a mutant of the Fny strain of CMV (Fny‐CMVΔ2b, which cannot express the CMV 2b protein) exhibit strong resistance against M. persicae, which is manifested by decreased survival and reproduction of aphids confined on the plants. Previously, we found that the Fny‐CMV 1a replication protein elicits aphid resistance in plants infected with Fny‐CMVΔ2b, whereas in plants infected with wild‐type Fny‐CMV this is counteracted by the CMV 2b protein, a counterdefence protein that, among other things, inhibits jasmonic acid (JA)‐dependent immune signalling. We noted that in nontransformed cv. Petit Havana SR1 tobacco plants aphid resistance was not induced by Fny‐CMVΔ2b, suggesting that not all tobacco varieties possess the factor(s) with which the 1a protein interacts. To determine if 1a protein‐induced aphid resistance is JA‐dependent in Xanthi tobacco, transgenic plants were made that expressed an RNA silencing construct to diminish expression of the JA co‐receptor CORONATINE‐INSENSITIVE 1. Fny‐CMVΔ2b did not induce resistance to M. persicae in these transgenic plants. Thus, aphid resistance induction by the 1a protein requires JA‐dependent defensive signalling, which is countered by the CMV 2b protein. [ABSTRACT FROM AUTHOR]

Seed lots of tomato and capsicum (Solanum lycopersicon and Capsicum annuum, respectively) are required to be free of quarantine pests before their entry to Australia is permitted. Testing of samples from 118 larger seed lots in the period 2019–2021 revealed that 31 (26.3%) carried one or more of four Tobamovirus species, including tomato mottle mosaic virus (ToMMV), which is a quarantine pest for Australia. Testing of samples from a further 659 smaller seed lots showed that 123 (18.7%) carried a total of five Tobamovirus species, including ToMMV and tomato brown rugose fruit virus (ToBRFV), which is also a quarantine pest for Australia. Estimated prevalence of contamination by tobamoviruses ranged from 0.388% to 0.004% in contaminated larger seed lots. Analyses of these data allow us to estimate probabilities of detection of contamination under different regulatory settings. [ABSTRACT FROM AUTHOR]

Pesticides are essential for the development of agriculture. It is urgent to develop green, safe and efficient pesticides. Bisindole alkaloids have unique and concise structures and broad biological activities, which make them an important leading skeleton in the creation of new pesticides. In this work, we synthesized bisindole alkaloid barakacin in a simple seven-step process, and simultaneously designed and synthesized a series of its derivatives. Biological activity research indicated that most of these compounds displayed good antiviral activities against tobacco mosaic virus (TMV). Among them, compound 14b exerted a superior inhibitory effect in comparison to commercially available antiviral agent ribavirin, and could be expected to become a novel antiviral candidate. Molecular biology experiments and molecular docking research found that the potential target of compound 14b was TMV coat protein (CP). These compounds also showed broad-spectrum anti-fungal activities against seven kinds of plant fungi. [ABSTRACT FROM AUTHOR]

Tomato mottle mosaic virus (ToMMV) poses a threat to production and quality of tomato fruits. The Tm-22 gene confers resistance to some tobamoviruses by recognizing viral movement proteins. However, Tm-22-mediated resistance against ToMMV is not well known. Here, we found that ToMMV could infect wild-type but not Tm-22 transgenic Nicotiana benthamiana plants and could also infect tomato cultivar Moneymaker but not resistant cultivar Jili with homozygous Tm-22. Chimeric viral ToMMVToBRFV−MP with swapped ToMMV MP to MP of tomato brown rugose fruit virus could systemically infect Tm-22 transgenic N. benthamiana and tomato cultivars Jili plants. Further, transient expression of ToMMV MP in the leaves of Tm-22 transgenic N. benthamiana plants induced hypersensitive response-associated cell death, suggesting that the MP of ToMMV was the avirulent factor for the Tm-22 resistance gene. ToMMV could infect Tm-22-containing cultivar Jinpeng 1 but not Chaobei. Sequence analysis revealed that cultivars Chaobei and Jinpeng 1 were heterozygous, where Chaobei consists of Tm-22 and Tm-2 genes, while Jinpeng 1 consists of Tm-22 and tm-2 genes. Transient co-expression assays showed that both Tm-22 and Tm-2 but not tm-2 could recognize ToMMV MP and induce hypersensitivity response-associated cell death in N. benthamiana leaves, suggesting that homozygous tomato harboring Tm-22 and heterozygous tomato containing Tm-22 and Tm-2 may exhibit more durable resistance to ToMMV than heterozygous tomato carrying Tm-22 and tm-2. Further, Tm-22 transgenic N. benthamiana and tomato cultivar Jili plants with silenced Tm-22 gene were susceptible to ToMMV. Also, silencing type-I J-domain MIP1 gene compromised Tm-22-mediated resistance to ToMMV in Tm-22 transgenic N. benthamiana and tomato cultivar Jili. Moreover, we found that viral RNA could accumulate in the systemic leaves of Tm-22 transgenic N. benthamiana plants and tomato cultivar Jili at 35°C, but not at 20, 25, or 30°C. Altogether, our findings reveal that the Tm-22 confers resistance to ToMMV by recognizing MP, and the resistance is regulated by the allele combinations, accumulation levels of Tm-22, MIP1, and the temperature. [ABSTRACT FROM AUTHOR]

Summary: Molecular farming technology using transiently transformed Nicotiana plants offers an economical approach to the pharmaceutical industry to produce an array of protein targets including vaccine antigens and therapeutics. It can serve as a desirable alternative approach for those proteins that are challenging or too costly to produce in large quantities using other heterologous protein expression systems. However, since cost metrics are such a critical factor in selecting a production host, any system‐wide modifications that can increase recombinant protein yields are key to further improving the platform and making it applicable for a wider range of target molecules. Here, we report on the development of a new approach to improve target accumulation in an established plant‐based expression system that utilizes viral‐based vectors to mediate transient expression in Nicotiana benthamiana. We show that by engineering the host plant to support viral vectors to spread more effectively between host cells through plasmodesmata, protein target accumulation can be increased by up to approximately 60%. [ABSTRACT FROM AUTHOR]