Atgstu19 and Atgstu24 knockout mutants to mechanical inoculation of Turnip mosaic virus (TuMV) was examined. The systemic infection of TuMV was more dynamically promoted in Atgstu19 mutants than in wild-type (Col-0) plants, suggesting the role of GSTU19 in TuMV resistance. However, Atgstu24 mutants displayed virus limitation and downregulation of the relative expression of TuMV capsid protein, accompanied rarely by TuMV particles only in vacuoles, and ultrastructural analyses of inoculated leaves revealed the lack of virus cytoplasmic inclusions. These findings indicated that Atgstu24 mutants displayed a resistance-like reaction to TuMV, suggesting that GSTU24 may suppress the plant resistance. In addition, these findings confirmed that GSTU1 and GSTU24 are induced and contribute to the susceptible reaction to TuMV in the Atgstu19–TuMV interaction. However, the upregulation of GSTU19 and GSTU13 highly correlated with virus limitation in the resistance-like reaction in the Atgstu24–TuMV interaction. Furthermore, the highly dynamic upregulation of GST and glutathione reductase (GR) activities resulted in significant induction (between 1 and 14 days post inoculation [dpi]) of the total glutathione pool (GSH + GSSG) in response to TuMV, which was accompanied by the distribution of active glutathione in plant cells. On the contrary, in Atgstu19, which is susceptible to TuMV interaction, upregulation of GST and GR activity only up to 7 dpi symptom development was reported, which resulted in the induction of the total glutathione pool between 1 and 3 dpi. These observations indicated that GSTU19 and GSTU24 are important factors in modulating the response to TuMV in Arabidopsis thaliana. Moreover, it was clear that glutathione is an important component of the regulatory network in resistance and susceptible response of A. thaliana to TuMV. These results help achieve a better understanding of the mechanisms regulating the Arabidopsis–TuMV pathosystem.Plants produce glutathione as a response to the intercellular redox state. Glutathione actively participates in the reactive oxygen species (ROS)-dependent signaling pathway, especially under biotic stress conditions. Most of the glutathione S-transferases (GSTs) are induced in cells during the defense response of plants not only through highly specific glutathione-binding abilities but also by participating in the signaling function. The tau class of GSTs has been reported to be induced as a response under stress conditions. Although several studies have focused on the role of the tau class of GSTs in plant–pathogen interactions, knowledge about their contribution to the response to virus inoculation is still inadequate. Therefore, in this study, the response of  NTN)–potato interaction [phi class in Solanum tuberosum, regardless of interaction type was reported, among which StGSTF2 acts as a marker of resistance response in hypersensitive reaction to PVYNTN.Glutathione (in reduced form—GSH) is a ubiquitous thiol in eukaryotes. GSH plays a vital role in plant development  and medieraction . Furthertau (GSTU) and phi (GSTF) classes being the largest groups [Arabidopsis thaliana belong to the GSTU class, gaps in knowledge about the functions of GSTUs in plant stress response still remain. Moreover, recent studies focus mainly on the role of GSTUs under abiotic stress conditions [A. thaliana to fungal pathogens Eryshiphe pisi, Colletotrichum, and Plectosphaerella [tau class member, is a part of the response to Verticillium dahliae. Although some information on the involvement of individual AtGSTUs in plant response to the pathogen is available, their role in plant–virus interactions is not adequately investigated. We chose Turnip mosaic virus (TuMV), which is the member of the largest family of RNA plant viruses—Potyviridae as a model in studies plant–virus interaction [Atgstu19 knockdown mutants have only been analyzed under salt treatments so far [Peronospora parasitica. Taking into account data presented by Wagner et al. [Peronospora parasitica isolate NOCO (compatible interaction) and P. parasitica isolate EMWA (incompatible interaction), expression of the AtGSTs was compared to the expression of the pathogenesis related protein (PR1), which can act as a marker for defense gene induction. In the compatible interaction, the expression of AtGSTU19  was found to be co-induced with PR1 three days after inoculation. Whereas in the incompatible interaction, the AtGSTFs showed much weaker or even no induction at all, while AtGSTU19 was transiently induced two days after infection, which preceded the induction of PR1 at 3 dpi. Therefore, we can surmise that AtGSTU19 can contribute in virus biotic stress as well, as we were intrigued how these plants would react to potyvirus inoculation.GSTs (E.C. 2.5.1.18) play the role of catalysts in GSH-dependent reactions, such as conjugation to substrates, hydroxyperoxidase activity, and dehydroascorbate reductase activity ,20. Sevet groups . Among tt groups ,24. Studt groups . GSTUs at groups . The expt groups , salinitt groups ,29,30. Tt groups ,31. Althnditions . GSTU13 haerella . Moreovehaerella  have reperaction . In thiseraction . Moreoves so far . Furthers so far . In addis so far . Howevers so far , who repr et al.  of inocuAtgstu24 mutant lines are redox homeostasis regulators as an effect of salt stress [Moreover, knowledge about AtGSTU24 under stress conditions is much poorer. GSTU24 transcripts are strongly induced by treatments with a wide range of xenobiotics ,42 and lt stress .Atgstu19 and Atgstu24 knockout mutants to TuMV pathogenic stress was examined. Significant differences were observed in the expression of the selected Atgstu genes, which indicated the specific contribution of individual AtGSTUs in different reactions to TuMV. Moreover, as the limitation virus content was highly dynamic, the induction of the total glutathione pool, accompanied by the upregulation of the activity GSTs and glutathione reductases (GRs), correlated with resistance-like reaction in the Atgstu24–TuMV interaction. On the contrary, significant changes in the distribution of glutathione in plant cells and glutathione pool induction of up to just 3 days post-inoculation (dpi) after TuMV infection showed susceptible Atgstu19–TuMV interaction. Moreover, these findings revealed the importance of selected AtGSTUs in the regulation of TuMV infection in Arabidopsis.In the present study, for the first time, the response of Atgstu19 and Atgstu24) were selected. Wild-type Col-0 displayed susceptibility to TuMV isolate PV-0104 [405 nm values were significantly higher in Atgstu19 plants than in Atgstu24 plants and susceptible Col-0 plants by 3 to 14 dpi  and also in Atgstu19 (4.47-fold between 3 and 7 dpi and 1.21-fold between 7 and 14 dpi) plants. However, in TuMV-inoculated Atgstu24 plants, the corrected mean OD405 nm values only indicated a significant increase in virus concentrations between 3 and 7 dpi (approximately 1.31-fold) and a high level of decrease in virus concentrations between 7 and 14 dpi  gene, which was analyzed based on two plant host reference genes—AtEf1α and AtF-Box—to show changes in TuMV concentrations in the inoculated leaves  and in Atgstu19 (2.48-fold between 3 and 7 dpi and 2.84-fold between 7 and 14 dpi) plants. Furthermore, the TuMV concentration and the dynamics of the increase in TuMV-CP expression were much higher in Atgstu19 plants than in Col-0 plants. However, in virus-inoculated Atgstu24 plants, TuMV-CP expression indicated an increase in virus concentration between 3 and 7 dpi (2.32-fold) and a high reduction in virus concentration between 7 and 14 dpi  between 7 and 14 dpi  experiments were carried out to analyze the levels of different glutathione forms, such as reduced glutathione—GSH  than in Col-0 plants. These results also revealed that 7 dpi is a crucial time point in the susceptible interaction between TuMV and Arabidopsis plants. All plants started showing problems in counteracting oxidative stress at 7 dpi. However, more severe problems in counteracting the oxidative stress were observed in the more susceptible Atgstu19 plants.The changes in the ione—GSH A, oxidizione—GSH B, and toione—GSH C after T to TuMV A. Moreovd 14 dpi A. Virus-Atgstu24 plants, the GSSG concentration steadily increased 1.77-fold between 1 and 14 dpi and was much higher than in mock-inoculated plants. This also suggested that TuMV inoculation modulated the GSH concentration and resulted in the direct conversion of GSH to GSSG. This observation underlined that virus-inoculated Atgstu24 plants showed precise protection against oxidative stress using upregulated levels of GSH, which was observed with the increased production of the GSSG form. However, in virus-inoculated susceptible Col-0 and Atgstu19 plants, the GSSG concentration was downregulated 4-fold and 4.29-fold, respectively. This demonstrated the problems in counteracting the oxidative stress in Col-0 and Atgstu19 plants. Further evaluation using HPLC analyses revealed that not only GSH and GSSG forms were modulated, but there were also changes in the total glutathione (GSH+GSSG) level  and TuMV–Atgstu19-susceptible plants, a slight upregulation was observed only between 1 and 7 dpi, with a strong reduction between 7 and 14 dpi—1.89-fold for Col-0 and 3.48-fold for Atgstu19.The concentration of GSSG was modulated during TuMV infection B. In virG) level C. In resAtgstu24 interaction, whereas in the TuMV–Atgstu19 and TuMV–Col-0 interactions, an increase up to the 7 dpi time point was observed. However, this observation did not take into account the subcellular distribution of glutathione in plant cell compartments, which could be important in response to TuMV.Significant changes in GSH and GSSG concentrations indicated that the glutathione content was highly increased, especially in the TuMV–Atgstu19, Atgstu24, and Col-0. Based on the glutathione content analyses, 7 and 14 dpi time points were selected to present the glutathione distribution changes at the ultrastructural level. Subcellular distribution confirmed that TuMV inoculation induced a higher glutathione deposition between 7 and 14 dpi in plant cells, compared with mock-inoculated plants, but the most dynamic induction in the resistance-like reaction was observed during the TuMV–Atgstu24 interaction  analyses, described as the pathogen reaction related to GSTU genes, such as AtGSTU1 and AtGSTU13 [Atgstu19 and Atgstu24 mutants, the expression levels of AtGSTU19 and AtGSTU24 were tested in Col-0, Atgstu19, and Atgstu24 plants as well  and Atgstu19 (1.27-fold) plants , and, compared with mock-inoculated plants (1.12-fold), it was drastically reduced between 7 and 14 dpi.The normalized expression of the ) plants A. HoweveAtGSTU13 expression was more significantly upregulated (1.43-fold) between 3 and 14 dpi, compared with mock-inoculated plants, and to the highest levels in the Atgstu24–TuMV interaction  between 3 and 14 dpi. However, in susceptible Col-0 plants, AtGSTU19 expression was upregulated only 1.1-fold between 3 and 7 dpi and highly downregulated (6.25-fold decrease). On the contrary, Atgstu24 plants showed stable and high induction of the AtGSTU19 gene during all analyzed time intervals (3.54-fold increase between 3 and 14 dpi). Moreover, the AtGSTU19 gene was also slightly induced in Col-0 plants between 3 and 14 dpi.The analyses of ve genes A,B. MoreAtGSTU gene expression showed that the resistance-like reaction was associated with virus limitation in the TuMV–Atgstu24 interaction and revealed the upregulation of AtGSTU13 and AtGSTU19 genes and the downregulation of AtGSTU1 gene. However, virus propagation susceptibility observed in the TuMV–Atgstu19 interaction was accompanied by the increased expression of AtGSTU1 and AtGSTU24 genes with the downregulation of AtGSTU13. Therefore, for an in-depth understanding of the relationship between Atgstu19 and Atgstu24 plants’ responses to TuMV inoculation, the correlation between the change of susceptibility/or resistance (based on TuMV concentration) and modulation of the AtGSTU gene expression  was evaluated, which is considered the marker for resistant/or even hypersensitive reaction to viral pathogens [Atgstu24 (resistance-like reaction) plants from 3 to 14 dpi, 1.55-fold and 1.2-fold, respectively (Atgstu19 (susceptibility) plants, they were induced only between 3 and 7 dpi, following which they significantly decreased at the 14 dpi time point  and mock-inoculated plants. Moreover, A. thaliana plants with a knockout of single GST from the tau group showed different responses to TuMV inoculation. In general, plant GSTs are postulated as enzymes involved in diverse functions, ranging from plant detoxification and ROS homeostasis to signaling molecules and adaptors in biotic stress [phi class StGSTF genes is differentially regulated during interactions between S. tuberosum cultivars and PVYNTN [StGSTF2 corresponded to HR induction and reduced PVYNTN concentration. Moreover, as reported in previous studies, StGSTF2 participated not only in resistance response but also in systemic infection in susceptible potato reaction. Therefore, we decided to examine whether the second largest GST class—tau—has an impact on plant–TuMV interactions. In Arabidopsis, the GSTU class includes 28 members [GSTU genes show enhanced stress resistance [A. thaliana gstu19 knockout mutants exhibited susceptible response to TuMV, which was confirmed by the induction of TuMV-CP expression, a higher TuMV concentration between 3 and 14 dpi, ultrastructural alterations with virus particles, and virus cytoplasmic inclusion depositions. Moreover, among the selected AtGSTU genes, AtGSTU1 and AtGSTU24 were highly induced between 3 and 14 dpi during TuMV infection, compared with not only mock-inoculated plants but also wild-type susceptible Col-0. Furthermore, in our studies AtGSTU13 was activated in a susceptible reaction only up to 7 dpi until symptoms appeared (7 dpi time point); therefore, it can be potentially possible that GSTU13 contributed in symptoms development in Agstu19-TuMV pathosystem. Moreover, AtGSTU13 was also dynamically activated (between 3 to 14 dpi) in an Atgstu24-TuMV resistance-like reaction. Furthermore, PCCs analyses indicated the potential role of AtGSTU13 with an increased expression level in the resistance-like reaction, when AtGSTU13 correlated with the reduction in virus concentration, in contrast to the susceptible reaction in Atgstu19. Therefore, AtGSTU13 can potentially act a dual role, contributing in resistance reaction, as well as a participant in symptoms development in susceptible reaction. These findings on AtGSTU13 are partially consistent with those of the report presented by Zhang and co-authors [Soybean mosaic virus (SMV) at both transcriptional and protein levels.This study explored for the first time the response of c stress ,47,48. Ac stress . Our pred PVYNTN . We conf members . Some st members . It is rsistance . Moreovesistance ,51. Howe-authors , in whicAtGSTU1 and AtGSTU24 expression was observed between 3 and 7 dpi in the susceptible TuMV–Atgstu19 interaction. Therefore, it can be postulated that AtGSTU1 and AtGSTU24 contribute to the susceptibility of A. thaliana to TuMV and the limitation of oxidative stress, whereas, similar to StGSTF2 in PVYNTN–potato interactions, AtGSTU13 is involved in the induction of virus symptoms in susceptible reactions.Moreover, a strong correlation between TuMV concentration and Obuda pepper virus [Rice stripe virus disease in Arabidopsis [Beta vulgaris–Beet necrotic yellow vein virus (BNYVV) interactions [Cucumber green mottle mosaic virus [A. thaliana to Cauliflower mosaic virus, systemic induction of GST1 with increased virus titers and development of symptoms was observed [Mungbean yellow mosaic India virus and Mungbean yellow mosaic virus. In addition, Skopelitou et al. [Bamboo mosaic virus–Nicotiana benthamiana interaction, confirming the ability of binding to the UTR region of (+) s virus RNA, leading to effective replication. The protein–protein interaction between host plants and pathogens is an active area of research that can not only widen our understanding of virus–plant interaction but also facilitate resistance breeding [Potyviridae family is a frequent feature during induction and support of virus infection in different host reaction [N. benthamiana–Colletotrichum destructivum and N. benthamiana–Colletotrichum orbiculare interactions, two GSTU genes were highly induced: NbGSTU1 and NBGSTU3. When the transcription of NbGSTU1 was blocked by gene silencing, the resistance to C. orbiculare was highly suppressed. Moreover, a 67% higher fungi colonization and a 130% higher number of lesions caused by C. orbiculare were observed, compared with mock-inoculated plants. Moreover, in A. thaliana, a tau class gene GSTU13 was identified as an indispensable component of the immune pathway, as the lack of functional GSTU13 resulted in enhanced susceptibility to the following fungal pathogens: Plectosphaerella cucumerina, Colletotrichum, or Erysiphe [GSTU genes may suppress the resistance of plants to pathogens. Therefore, we postulated that GSTU19 in the A. thaliana–TuMV interaction can act as a crucial factor for the resistance of plants to TuMV.Interestingly, transcriptomic analyses revealed that the GST expression profile can be differentially regulated in plant–virus interactions, such as pepper leaves infected with er virus , Rice stbidopsis , Beta vuractions , and resic virus . Moreoveobserved . Furtherobserved  observedu et al.  reportedu et al.  observedbreeding . The abireaction ,60. In treaction  reportedErysiphe . SimilarAtgstu24–TuMV pathosystem. In Atgstu24 mutants, virus titers quantified by DAS-ELISA were lower between 3 and 14 dpi, which was confirmed by the decrease in the relative expression of TuMV-CP, especially between 7 and 14 dpi, compared with Col-0 and Atgstu19. Moreover, ultrastructural analyses with virus capsid immunogold labeling indicated that in the TuMV–Atgstu24 interaction, virus particles were observed in lower frequency and only in vacuoles. Furthermore, in these interactions, neither the induction of virus cytoplasmic inclusion typical for Potyvirus susceptible reaction nor chloroplast lamella disorganization [Atgstu24–TuMV pathosystem, higher levels of virus limitation and resistance-like reaction were observed, compared with wild-type reaction and, in particular, Atgstu19. This was accompanied by a significant reduction in TuMV-CP deposition between 7 and 14 dpi. In the present study, relative expression analyses of the selected AtGSTU genes indicated that AtGSTU13 and AtGSTU19 were highly induced between 3 and 14 dpi. Moreover, the dynamic increase in AtGSTU19 and AtGSTU13 expression strongly correlated with the reduction in virus concentration, in contrast to the susceptible reaction in Atgstu19. Furthermore, in Atgstu24 response to TuMV, GSTU1 was also induced, only up to the time point when infection symptoms appeared at 7 dpi, whereas it was highly reduced between 7 and 14 dpi. We postulated that in the response of Atgstu24 to TuMV, GSTU19 and GSTU13 contribute to resistance-like reaction induction, whereas GSTU1 contributes to symptom development. The induction of AtGSTU19 and AtGSTU13 in Arabidopsis response to TuMV was similar to the activation of StGSTF2 in the hypersensitive response of potato to PVYNTN, which correlated with PVY-CP downregulation [NtGSTU1 was observed between 3 and 6 h after virus inoculation, causing a reduction in TMV replication [Barley mosaic virus and Potato virus X but had no influence on Cucumber mosaic virus. Satoh et al. [Rice tungro spherical virus of rice cultivars. Wang et al. [Triticum aestivum has an important role in resistance to Bgt (Blumeria graminis f. sp. tritici) but not to Pst (Puccinia stitiformis f. sp. tritici). The overexpression of TaGSTU6 in Arabidopsis induced resistance to Pseudomonas syringae pv. Tomato DC3000 [A. thaliana during Alternaria brassicicola infection [GSTU1, GSTU11, and GSTU10 genes were actively induced two days after fungus inoculation. Glutathione transferases are known for their role in maintaining the physiological redox state of the plant cell [Atgstu19 and TuMV–Atgstu24 interactions. Numerous studies have shown that the induction of the glutathione content is important for plant disease resistance response. For example, Király et al. [Atgstu24–TuMV interaction: the highly dynamic increase in the glutathione form between 1 and 14 dpi, accompanied by a significant increase in the GSSG form. Moreover, the levels of GSH and GSSG were the highest in Atgstu24–TuMV interactions, compared with Atgstu19–TuMV and Col-0–TuMV interactions. These observations were quite different from those presented by Horváth et al. [Atgstu19 knockdown line and Atgstu24 knockout plants under salt stress conditions. However, plants that underwent Atgstu24 knockout mutant salt treatment showed an increased content of total glutathione and a high GSH/GSSG ratio. A similar tendency to Atgstu24–TuMV interactions was observed in PVYNTN–resistant potato Neptun [NTN inoculation between 1 and 21 dpi, compared with susceptible reactions and mock-inoculated plants. These findings are consistent with those of Singh et al. [Quite a different scenario was observed in the nization  was obsegulation . As repogulation , GST in lication . Furtherlication . As obselication , GSTU4 dh et al.  postulatg et al.  reportedo DC3000 . In addinfection . GSTU1, ant cell . Severalant cell . Therefoy et al.  demonstry et al. . The finh et al. , where to Neptun . Higher h et al. , who reph et al.  and Künsh et al. , a high h et al. ,73.Atgstu19–TuMV and Col-0–TuMV interactions. Moreover, these observations are consistent with the data obtained in susceptible PVYNTN–potato Irys reaction [Atgstu19–TuMV interaction, the GSH content was highly induced up to 7 dpi, whereas the GSSG pool was only induced at the 1 dpi time point. Moreover, in susceptible Atgstu19–TuMV interactions, the total glutathione content (GSH + GSSG) increased up to 3 dpi, whereas it increased only between 3 and 7 dpi in Col-0 plants. Importantly, the decrease in the GSSG form started earlier (at 3 dpi) and was more dynamic than in Col-0. Finally, in susceptible Atgstu19–TuMV and Col-0, the GSH and the total glutathione content dynamically decreased at the 7 dpi time point, especially in Atgstu19–TuMV to a level even lower than in mock-inoculated wild-type Col-0. Considering the dynamic nature of the glutathione content during Col-0 and Atgstu19 induction in response to virus inoculation, TuMV-CP expression induction, and the tendency of virus titers, it can be concluded that the Atgstu19–TuMV interaction was more enhanced for susceptibility reaction than in Col-0 plants. This was also confirmed by ultrastructural alterations visualized by transmission electron microscopy and TuMV-CP deposition between Atgstu19–TuMV and Col-0 interactions [N. benthamiana susceptible to pepper mild mottle virus. It can be assumed that in line with Hernandez et al. [Significant differences in the glutathione concentration were observed in reaction . During ractions . A similractions , where tz et al. , glutathA. thaliana leaf tissues also revealed significant differences in the deposition between mock- and virus-inoculated plants and between Atgstu19 and Atgstu24 interactions. In mock-inoculated Col-0 plants, the most intense GSH deposition was detected in the following order: chloroplasts, mitochondria, and cytoplasm, similar to Atgstu19-mock-inoculated plants. In Atgstu24-mock plants as well, the deposition was the highest in chloroplasts, but quantification of gold granules indicated the highest GSH deposition in vacuoles, followed by chloroplasts. In general, gold labeling confirmed the glutathione content tendency in response to the virus in all interactions observed by HPLC analyses. Therefore, in Atgstu24 interactions, a steady increase in GSH localization was observed. On the contrary, in Col-0 and Atgstu19 interactions, localization was induced only up to 7 dpi, whereas an active decrease in localization was observed between 7 and 14 dpi, compared to mock-inoculated and to Atgstu24. Indeed, the highest levels of glutathione localization were reported in Atgstu24–TuMV. In Atgstu24–TuMV, the highest deposition was observed in the chloroplasts (much higher than in mock-inoculated Atgstu24), followed by the cytoplasm and the nucleus. The increase in the glutathione content in chloroplasts is an important factor for controlling ROS and symptom development. The findings of this study are consistent with those presented by Zechmann [Plum pox virus. Moreover, individual genes from Plum pox virus and TuMV expressed in plants can repress or change the virulence of virus to plants [Atgstu19 plants could disturb the natural defense process located in specific cell regions and promote viral infection. Moreover, in the Atgstu24–TuMV interaction, a highly induced GSH localization at the 7 dpi time point and a steady increase between 7 and 14 dpi in all cell compartments, except for mitochondria, were observed when the GSH content significantly decreased. Király et al. [Botrytis cinerea. Moreover, the lower level of glutathione in mitochondria may lead to its dysfunction and activation of the resistance response to the pathogen. Furthermore, only in the resistance-like reaction in Atgstu24–TuMV was a statistically steady stable significant cell wall deposition (7–14 dpi) observed. However, in virus-inoculated Col-0 and Atgstu19, cell wall localization was detected, but at the 7 dpi time point, it decreased to a nonsignificant level. As previously postulated by Tolin et al. [Atgstu19 and Col-0 inoculated tissues, the highest glutathione induction at 7 dpi was observed in the following order: nucleus, mitochondria, chloroplasts, and vacuoles. After symptom development between 7 and 14 dpi, a significant reduction in localization was noticed in all cell compartments in both interactions. The tendency of glutathione localization in the nucleus and the cytoplasm is quite similar to data reported in PVYNTN–potato interactions [The ultrastructural analyses of glutathione localization in inoculated Zechmann , where pZechmann , who preo plants . Becauseo plants  postulato plants  and Garco plants  suggestey et al.  and Simoy et al.  reportedn et al.  and Vanan et al. , the GSHractions . Glutathractions ,83,84. MGSTU genes were differentially expressed in Atgstu19–TuMV and Atgstu24–TuMV pathosystems, which was followed by significantly differentially changed glutathione reduced and oxidized forms and was accompanied by changes in GST and GR activities. Gullner et al. [Sugarcane mosaic virus (ScMV). They observed a more than 50% increase in the GST activity in the first 3 dpi in ScMV-resistant response, whereas susceptible cultivars exhibited a decreased GST activity. Moreover, they postulated that the GST activity may be associated with the resistance response to the virus. An increase or decrease in the GST activity can be a marker of susceptibility. Therefore, strong induction in Atgstu24–TuMV interaction 3–14 dpi was correlated with the resistance-like reaction, whereas weaker induction only between 3 and 7 dpi and a decrease between 7 and 14 dpi were associated with susceptibility. Moreover, Atgstu19 knockout line–TuMV interaction revealed a lower GST activity than Col-0-virus-inoculated and Col-0-mock inoculated plants. Col-0 exhibited a less susceptible reaction than Atgstu19 to TuMV. Horváth et al. [Atgstu19 knockdown plants treated with salt stress. Similarly, a long-term systemic infection caused by Plum pox virus revealed a strong decrease in the GST and GR activities [Atgstu19. Virus-inoculated Col-0 plants revealed a higher GST activity than in Atgstu19–TuMV interaction. However, in Atgstu19–TuMV, a lower activity was observed than in the resistance-like reaction in Atgstu24–TuMV. Moreover, the highest activity was reported in Atgstu24–TuMV between 3 and 14 dpi, compared with mock- and all virus-inoculated plants. Fodor et al. [White clover mosaic virus (WCMV)–bean interactions 10 dpi virus inoculation. Li and Burrit [Dactylis glomerata–Cocksfoot mottle virus (CfMV) interaction between 3 and 5 dpi. Amari et al. [Prunus necrotic ringspot virus (PNRSV)–apricot interaction. The pool of reduced glutathione GSH may be fueled by the GR activity. GR catalyzes the reduction of glutathione disulfide (GSSG) to two molecules of GSH, and thus electron transfer from NADPH and is crucial for maintaining the glutathione redox potential in different plant cell compartments.In our experiments, several r et al.  and Wu er et al.  studied h et al.  reportedtivities . In addir et al.  reportedr et al.  reportedd Burrit  reportedi et al.  reportedArabidopsis thaliana (L.) Heynh wild-type (Col-0) plants and A. thaliana lines containing a T-DNA insertion were used: in AtGSTU24  compensation of mutation in Arabidopsis glutathione transferase (AtGSTU) genes under control or salt stress conditions and in AtGSTU19 . Athgstu24 are knockout mutants, as reported by Horváth et al. [Atgstu19 is also a knockout mutant. All homozygous mutant seeds were kindly provided by Jolán Csiszár Laboratory. For TuMV inoculation described below, 18-day-old plants without any lesions and/or alterations were used. Healthy and mock-inoculated Athgstu24 and Athgstu19 revealed phenotype differences  using the Statistica software , as described by Kozieł et al. [405 nm values were computed as presented in a previous study [ł et al. , Tomilsoł et al. , and Walł et al.  using thł et al. . InteresAtgstu24 . Leaves ł et al. . For a mus study  and usedus study  and presus study .405 nm were compared to the calculated cutoff point, and all OD405 nm values greater than 0.1412 were considered positive (confirmed presence of virus) [A. thaliana plants. Moreover, to double-check the level of TuMV, qPCR of the TuMV-CP gene fragment was performed using the primers presented by Arous et al. [AtEf1α : At5g60390) and AtF-Box : At5g15710), as presented by Lilly et al. [TuMV-CP gene. For DAS-ELISA, TuMV-CP expression, and other analyses , 50 plants were used (25 virus-inoculated and 25 mock-inoculated of each Col-0 and two glutathione transferase mutants). The analyses were performed in triplicate using a new set of plants every time.This calculated cutoff point was 0.1412. The readings of ODf virus) . Signifis et al. , and they et al. . The levA. thaliana GSTU genes in the plant host, molecular analyses were performed on the samples collected at 3, 7, and 14 dpi. Briefly, leaf samples (0.1 g of each sample) were collected from 25 mock- (buffer) or virus-infected plants of different types of Col-0 and mutant plants. RNA isolation, purification, and quality analyses were carried out following previously described procedures [AtEf1α  and AtF-Box  as reference standards [To estimate the expression of ocedures ,99,100. tandards , which cAtEf1α and AtF-Box reference genes. All qPCR tests were calibrated using previously prepared 5-point calibration curves (based on cDNA and gDNA). The following genes were selected based on pathogen reaction involvement [A. thaliana GSTU1  and GSTU13 . In addition, GSTU19  and GSTU24  were analyzed using qPCR expression. These host genes encoded protein products that were associated with the utilization of glutathione in response to stress (GSTU) [A. thaliana was analyzed, and complete sequences were acquired from the TAIR database [AtEf1α and AtF-Box as reference standards, as previously described [AtGSTU1, AtGSTU13, AtGSTU19, and AtGSTU24 and concentration of TuMV (based on the expression of TuMV-CP), correlation analyses were performed. To compare/check the likelihood between the expression of different GSTU genes and the level of the virus, PCCs were estimated according to Wu et al. [GSTU gene changes and levels of TuMV were estimated at 3, 7, and 14 dpi in Col-0 and mutant plants. The results were presented in the form of a heat map generated using PCC values, and values higher than 0.68 were considered to reflect the strong positive correlation between analyzed pairs.A real-time qPCR was performed using the Bio-Rad CFX96TouchTM apparatus  and Fast SG qPCR Master Mix (2×)  for olvement ,33,44 ans (GSTU) . The expdatabase . Moreoveescribed . The priescribed ,97. All u et al.  and Mandu et al.  by usingAtgstu19, and Atgstu24 plants were measured by reversed-phase HPLC with fluorescence detection, as reported by Kranner [The GSH and GSSG contents in mock- and TuMV-inoculated Col-0,  Kranner , using t Kranner . They weAtgstu19, and Atgstu24 leaves were collected at the 3, 7, and 14 dpi time points after the inoculation of mock or TuMV. The GST activity was validated as described by Islam et al. [−1 cm−1. To validate the enzymatic activity, 50 plants (25 virus-inoculated and 25 mock-inoculated of each Col-0 and two glutathione transferase mutants) were used. All analyses were performed in triplicate using a new group of plants every time.To validate the GST and GR activity, Col-0, m et al.  and Otulm et al. , and them et al. . The resm et al. . The GR 2) per image. For each combination , gold particles from 200 photographs were counted to determine the presence of glutathione or TuMV content.To analyze the virus concentration using microscopic studies, the leaf samples of mock- and virus-inoculated potato plants at the 7 and 14 dpi time point were embedded and treated following the procedure of Zechmann et al.  and KolbAtgstu19 and Atgstu24 knockout mutants to TuMV inoculation was examined and compared. Even though the general function of GSTUs has been documented, the potential function of AtGSTU-mediated response in TuMV remains unexplored so far.Many works have demonstrated that glutathione and glutathione metabolism enzymes play an important role under different plant stress conditions, especially pathogen challenges. However, glutathione metabolism in specific plant–virus pathosystems can be differentially modulated, and our knowledge about this is far from sufficient. Therefore, in this work, the response of Atgstu24–TuMV interactions, a more intense reduction in virus titers and a significant reduction in the TuMV-CP relative gene expression level were documented, compared with Col-0–TuMV and Atgstu19–TuMV. Ultrastructural analyses confirmed the localization of rare virus particles in vacuoles of inoculated leaf tissues and the lack of virus cytoplasmic inclusions and organelle alterations. Therefore, we postulated a resistance-like reaction to TuMV in Atgstu24, followed by a dynamic increase in the reduced GSH and total glutathione contents with high GST and GR activity. Importantly, in Atgstu24–TuMV, AtGSTU19 and AtGSTU13 contributed to a resistance-like reaction, whereas AtGSTU1 only participated in the early step of interaction until the symptoms appeared. Furthermore, glutathione activated plant defense, reduced the virus content, and decreased the potential damage to the host plant cell. Additionally, when a resistance-like reaction was induced in GSTU24 knockout mutants, AtGSTU24 may suppress plant resistance.In Atgstu19–TuMV interactions, induction of virus infection typical for the susceptible reaction was observed. Moreover, AtGSTU1 and AtGSTU24 highly correlated with susceptibility, but AtGSTU13 with symptom development at the 7 dpi time point, similar to Col-0 wild-type. Furthermore, the GSH content was only upregulated until 7 dpi and the total glutathione content until 3 dpi. However, the GSSG content decreased earlier and quicker (at 3 dpi), compared with Col-0, accompanied by GST and GR activity downregulation between 7 and 14 dpi. Therefore, taking into account much more intense virus content and glutathione modulation, enhanced susceptibility was observed in the Atgstu19–TuMV pathosystem. Additionally, when GSTU19 knockout mutants revealed susceptible reaction, resistant plants may require AtGSTU19. A comparison of A. thaliana knockout mutant–TuMV interaction indicated that different GSTUs can be involved in the differential modulation of plant response to TuMV inoculation. Further molecular and cellular studies on overexpressing GST mutants are needed to elucidate the possible role of other active components in the TuMV–A. thaliana pathosystem.On the contrary, in


Phage & Host Daily), a web application that offers a comprehensive, up-to-date catalog of known phage-host associations that allows users to select viruses targeting specific bacterial and archaeal taxa of interest. Our service combines the latest information on virus-host interactions from seven source databases with current taxonomic classification retrieved directly from the groups and institutions responsible for its maintenance. The web application also provides summary statistics on host and virus diversity, their pairwise interactions, and the host range of deposited phages. PHD is updated daily and available at http://phdaily.info or http://combio.pl/phdaily.Understanding phage-host relationships is crucial for the study of virus biology and the application of phages in biotechnology and medicine. However, information concerning the range of hosts for bacterial and archaeal viruses is scattered across numerous databases and is difficult to obtain. Therefore, here we present PHD ( Phages play a pivotal role in many ecosystems by shaping the structure of bacterial communities . They arBacillus megaterium” instead of Priestia megaterium) or misspelled . These issues have been addressed in two excellent databases, Virus-Host DB  such as RefSeq  or GenBa-Host DB  and NCBI-Host DB , both of-Host DB  and anno-Host DB . The MVP-Host DB . ConsequPhage & Host Daily), a daily updated web application that combines information on phage-host interactions from seven sources — NCBI Virus, Virus-Host DB, MVP, RefSeq, GenBank, UniProt, and IntAct. PHD provides information on hosts for prokaryotic viruses at the species level using two alternative taxonomic classification systems, NCBI Taxonomy  is extracted from assembly report files. Nucleotide sequences of viruses present in GenBank or RefSeq but not in the Assembly database are retrieved in FASTA and flat-file formats from NCBI Virus  taxonomy tables are downloaded from the NCBI FTP server. The ICTV taxonomy of viruses is retrieved from the Virus Metadata Resource at the ICTV website, and species are mapped to the corresponding NCBI taxonomy identifiers based on the RefSeq/GenBank genome accessions provided by ICTV. The GTDB taxonomy of Bacteria and Archaea is obtained from metadata files provided in the latest GTDB release. The bacterial and archaeal lineages are mapped between NCBI and GTDB taxonomies based on the NCBI taxonomy identifiers provided in the GTDB archaeal and bacterial metadata files.Virus-host assignments are retrieved from: (i) the NCBI Virus website, (ii) the TSV file provided by VirusHost DB, (iii) the text files from MVP, (iv) the GenBank flat files , (v) the protein-protein interactions from the IntAct FTP server, and (vi) the protein sequence entries in UniProt-SwissProt (“OH” line in UniProt entry). The extracted names and taxonomy identifiers of hosts are queried against NCBI Taxonomy using TaxonKit v. 0.10.1  to retriFor a prokaryotic virus infecting only one host species, the host range is set to this species. For a virus infecting multiple host species, we defined the host range as the taxonomic rank of the last common ancestor of all its hosts in the NCBI taxonomic database.The PHD web interface was developed in React.js (v. 17.0.2), Next.js (v. 11.1.3) and Highcharts.js (v. 10.0.0). The database querying system was developed in Django (v. 4.0.0), Django REST framework (v. 3.13.1), and Python (v. 3.9.5) using SQLite database as a management system.Siphoviridae, Myoviridae, and Podoviridae families  have been classified by ICTV, indicating a significant delay between NCBI submissions and classification by the committee. However, the number of taxa at higher ranks, from genus to phylum, is similar between NCBI and ICTV taxonomies . Both syfamilies . These ufamilies . Aside fTubulavirales (n = 71), Petitvirales (n = 67), and Haloruvirales , and lower taxonomic units not classified at the order level (n = 3). RNA viruses correspond to fewer than one percent of virus species (n = 27) and belong to five families: Cystoviridae (n = 16), Leviviridae (n = 5), Fiersviridae (n = 4), Steitzviridae (n = 1), and Duinviridae (n = 1).Consequently, most sequences in the database come from double-stranded DNA (dsDNA) viruses . Single-Escherichia virus G4/Gequatrovirus G4 (n = 343) and Escherichia virus phiX174/Sinsheimervirus phiX174 (n = 105). In both cases, the majority of retrieved sequences represents strains obtained during in vitro evolution experiments  have complete genomes, and the remaining viruses are represented by genomic fragments  or partial genomes at the contig and scaffold levels  (Most virus species (92%) are represented by single genome assembly. The remaining species mainly have two (4%) or three (1%) genomes assigned. The highest number of genomes have been reported for two closely related species, eriments . Over 92 n = 44) . Most viCampylobacter phage C10 is the shortest phage genome sequence  submitted to NCBI, the record itself (accession: MG065651) is flagged by the GenBank staff as “unverified”. The second smallest phage genome  belongs to Leuconostoc phage L5, which is often cited as the phage with the smallest known genome  have been isolated from Prevotella species .The size of complete genomes varies between 1.4 and 551.6 kb, with no homogenous distribution , which mn genome . At the n genome , there an genome . Such ph> 500 kb , n = 12)Mycolicibacterium, Escherichia, Streptococcus, Vibrio, Pseudomonas, Salmonella, Klebsiella, and Staphylococcus)  and 2.7% archaeal species  reported in NCBI Assembly  and 0.5% of archaeal species , respectively. Although collectively, host species represent 34 classes, three-quarters of the host species fall into five classes   . One of megmatis . In totaAssembly . Compareacteria) . Given tacteria) , these gn = 11,640) were isolated from single hosts, followed by viruses infecting two host species  . The remaining virus species  were reported to infect more than two host species   mostly f species . The recMost assignments between viral and host species were retrieved from NCBI Virus (93%) and GenBank (87%), followed by Virus-Host DB (35%) and RefSeq (31%) , indicatPhage & Host Daily (PHD) offers two ways to access information on interactions between virus and host species: by searching for a particular virus/host taxon and browsing taxonomic trees.The Search view allows users to look for viruses targeting bacterial or archaeal taxa of interest or prokaryotic taxa that are infected by phages from a given viral taxon. The view allows for searches corresponding to the names and identifiers used in NCBI, GTDB, and ICTV taxonomies. For convenience, the search box features an autocomplete functionality that suggests terms matching the user query. The Browse view provides a hierarchical exploration of virus-host interactions through virus or host taxonomies based on NCBI or GTDB Taxonomy. The interactive interface allows users to expand branches of virus or host trees and view the number of virus-host interactions associated with each node.Enterobacterales that are supported by RefSeq and Virus-Host DB and contain viruses with complete genomes).Once the query taxon is selected from either the Search or Browse view, PHD presents a table of pairwise interactions between viral and host species belonging to the query viral/host. For each virus-host interaction, PHD lists the source database(s), taxonomic affiliations for both viruses and hosts, as well as information on the virus’ genome composition and assembly completeness of the representative virus genome. This is a central component of PHD that can be filtered using multiple combinations of parameters  provides a single, convenient interface that allows for rapid access to an exhaustive set of experimentally verified phage-host interactions and provides up-to-date taxonomic classifications for all phages and hosts. We hope that our service will become a convenient one-stop-shop for biologists and bioinformaticians interested in finding novel, alternative hosts of known phages, spotting the bacterial taxa that might be neglected during earlier studies, and interpreting ecological relations observed in the environment. It can also be used by developers of bioinformatic tools to compile well-annotated phage and host datasets for their tools. Finally, our data can help to uncover links between genomics and the phylogeny of prokaryotic viruses, and their host range.The original contributions presented in this study are included in the article/AZ conceived and supervised the project. AZ and AR implemented the database and methods for data collection. KA designed and implemented the user interface. AZ and MH prepared the figure. AZ, MH, and JB analyzed the data and wrote the manuscript. All authors reviewed and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Ipomoea batatas), a staple food for people in many of the least developed countries, is affected by many viral diseases. In 2017, complete genome sequences of sweet potato symptomless virus 1  isolates were reported, although a partial SPSMV-1 genome sequence had previously been identified by deep sequencing. To assess the presence of this virus in Spain, sweet potato leaf samples collected in Málaga  and the Spanish Canary Islands of Tenerife and Gran Canaria were analyzed. SPSMV-1 was detected in samples from all the geographical areas studied, as well as in plants of several entries obtained from a germplasm collection supposed to be virus-free. Sequence analysis of full-length genomes of isolates from Spain showed novel molecular features, i.e., a novel nonanucleotide in the intergenic region, TCTTATTAC, and a 24-nucleotide deletion in the V2 open reading frame. Additionally, an agroinfectious clone was developed and infectivity assays showed that the virus was able to asymptomatically infect Nicotiana benthamiana, Ipomoea nil, I. setosa, and sweet potato, thus confirming previous suggestions derived from observational studies. To our knowledge, this is the first report of the presence of SPSMV-1 in Spain and Europe and the first agroinfectious clone developed for this virus.Sweet potato ( Ipomoea batatas) is one of the most consumed crops in the world and is a staple food for people in many of the least developed countries. This crop is the sixth most important food crop worldwide after rice, wheat, potato, maize, and cassava. Sweet potato crops produce more edible energy per hectare per day than other important crops, such as wheat, rice, and cassava, and are an important source of carbohydrates, beta-carotene, and vitamins B, C, and E. Sweet potato is vegetatively propagated by vine cuttings, which cause the accumulation of pathogens, mainly viruses. Consequently, this crop is particularly affected by viral diseases that substantially decrease the quality and yield of the storage roots [Sweet potato  sweet potato chlorotic stunt virus [Potyvirus, family Potyviridae) sweet potato feathery mottle virus, sweet potato virus G, and sweet potato virus 2 [Begomovirus, family Geminiviridae) sweet potato leaf curl virus and sweet potato leaf curl Canary virus [Deltasatellite, family Tolecusatellitidae) associated with the begomoviruses infecting sweet potato has also been reported in this country [In Europe, the demand for sweet potatoes has greatly increased in recent years, leading to an increase in imports and production in southern European countries, including Spain. Although extensive field studies of the losses caused by sweet potato viruses have not been carried out in Spain, several viruses have been reported to infect this crop. They are the crinivirus  icosahedral particles [The genus iviridae . Mastrevarticles . Mastrevarticles .In 2009, the partial genome sequence of a mastrevirus was identified in sweet potato by the deep sequencing of small RNAs in Peru . Until nNicotiana benthamiana, Ipomoea nil, I. setosa, and sweet potato with no noticeable symptoms.In this study, the presence of SPSMV-1 was assessed for the first time in Spain, with the virus found in samples from all of the geographical areas studied. This is the first record of the virus in Europe. SPSMV-1 was also detected in several samples of pathogen-tested in vitro plants obtained from a germplasm collection. In addition, the first agroinfectious clone of the virus was developed, and infectivity assays showed that it was able to infect I. batatas) (n = 71) from Málaga, Tenerife, and Gran Canaria and ornamental or naturalized I. indica (n = 24) from Tenerife and Gran Canaria. Most of the plants did not show any conspicuous symptoms.Leaf samples were collected from 95 plants from Málaga province  and the Spanish Canary Islands of Tenerife and Gran Canaria . SampledTotal DNA was extracted from leaf samples using a cetyltrimethylammonium bromide (CTAB)-based purification method . PCR to http://www.ncbi.nlm.nih.gov/Blast.cgi (accessed on 1 May 2022)) was used to perform the sequence similarity search. Pairwise identity scores were calculated with the Sequence Demarcation Tool (SDT) [Sequences were analyzed with the Lasergene sequence analysis package . The BLASTn algorithm was used to determine sequence similarity in the GenBank database. The BLASTn program  (http://ol (SDT)  after seol (SDT) . The besol (SDT) .GGTACCGTGTATTTGATGACGATGTAC)/MA27784 (GGTACCCCCTGGGTTGAACACAAC) were designed to amplify the full-length genome of an isolate of SPSMV-1 present in sample B2 from Málaga (GenBank accession numbers ON526997). These primers included the sequence of the KpnI restriction site (underlined) naturally present in the viral genome sequence. PCR was carried out as follows: denaturation for 3 min at 95 °C, 34 cycles of denaturation for 1 min at 95 °C, hybridization for 1 min at 66 °C, and extension for 3 min at 72 °C, followed by a final extension step of 5 min at 72 °C. The infectious dimeric SPSMV-1 clone was constructed essentially as described by Ferreira et al. [KpnI from the pGEM-T-Easy Vector, religated, and used as a template for rolling circle amplification (RCA) . The RCA product was partially digested with KpnI to produce dimeric molecules that were cloned in the pUC18 vector. The insert of the dimeric clone was excised with BamHI/Eco53KI and subcloned in the BamHI/SmaI sites of the pCAMBIA0380 binary vector.Primers MA2782  liquid medium containing kanamycin (50 µg/mL) and rifampicin (50 µg/mL). For agroinoculation assays, A. tumefaciens cultures were used to inoculate plants by stem puncture, as previously described [A. tumefaciens C58C1 cultures containing the empty pCAMBIA0380 vector were used as negative controls (mock). Nicotiana benthamiana was inoculated at the four-leaf stage. Ipomoea nil and I. setosa were inoculated at the two-leaf stage and cuttings of sweet potato cv. ‘Tanzania’ and ‘Camote Morado’ were inoculated. Two independent experiments were carried out for each plant species. Plants were maintained in an insect-free growth chamber  until analyzed.An escribed ,21. PlanI. indica plants from Málaga  and the Spanish Canary Islands of Tenerife and Gran Canaria was analyzed for the presence of SPSMV-1. PCR with primers Detect-1F/Detect-1R showed the expected DNA fragment of 418 bp in sweet potato samples from all locations  (I. indica samples showed the expected DNA fragments (The DNA extracted from the leaf samples of sweet potato and  Málaga) A. Overalragments .Sweet potato plants of cultivars ‘Tanzania’, ‘Blanca,’ ‘Beauregard,’ ‘Promesa’, and ‘Camote Morado’ obtained from the USDA-ARS Plant Genetic Resources Conservation Unit  and maintained by cuttings in insect-proof chambers for more than 10 years were screened with primers Detect-1F/Detect-1R to detect SPSMV-1. Surprisingly, the virus was detected in cultivars ‘Blanca’, ‘Beauregard’, and ‘Promesa’ B. To ensTo molecularly characterize SPSMV-1 isolates from Spain and establish the phylogenetic relationships with isolates from other countries, the full-length viral genome was amplified with primers Full-4F/Full-4R from samples CI24 from Tenerife; CI61 from Gran Canaria; and B2, B3, and B14 from Málaga. The expected DNA fragments corresponding to the full-length SPSMV-1 genomes were cloned into the pGEM-T-Easy Vector. Two clones were sequenced per sample and deposited in the GenBank database under accession numbers ON526993–ON527002.′ end of V2 ORF. The movement protein encoded by the V2 ORF in the mentioned isolates lacked the corresponding eight amino acids present in all previously sequenced SPSMV-1 isolates deposited in GenBank.Sequences obtained in this work showed a similarity of 99.0–99.8% and at least 96.9% similarity with all SPSMV-1 isolates available in GenBank . The unuPhylogenetic analysis of all SPSMV-1 isolates available, including those obtained in this work, showed no correlation between the isolates and their geographic origin .N. benthamiana, I. nil, and I. setosa; and the natural host sweet potato (cv. ‘Tanzania’ and ‘Camote Morado’). Plants inoculated in two independent experiments and analyzed by nested PCR with primers MA2924/MA2925 and MA2926/MA2927 at 5 weeks post-inoculation showed that the virus was able to infect all of the plant species assayed . No evident symptoms of the disease were observed in any of the infected plants. Thus, our experimental results support previous observations that SPSMV-1 could not be associated with any conspicuous symptoms in sweet potato plants [In this study, we developed for the first time an agroinfectious clone for SPSMV-1. Our results showed that the clone was able to infect the model plant o plants ,10. Howeo plants ,23. The o plants ,24.In summary, SPSMV-1 was confirmed as an asymptomatic mastrevirus infecting sweet potato crops in an increasing number of countries including Spain. Several biological aspects remain to be elucidated about this virus, including the possible interactions with other viruses in mixed infections and the mode of transmission. The putative insect vector is likely a leafhopper, similar to other mastreviruses .


Tick-borne diseases (TBDs) are bacterial, viral, and parasitic diseases transmitted by ticks. Viral TBDs have increased in prevalence over the last decade with many new pathogenic viruses being discovered. Doxycycline is often empirically prescribed by clinicians to treat symptomatic patients following tick bites due to suspicions of bacterial TBDs such as Rocky Mountain spotted fever, anaplasmosis, and ehrlichiosis. However, viral TBDs are included in the differential diagnosis if patients do not clinically improve following antibiotic therapy. Several viral TBDs present with dermatological manifestations. Recognizing the differences in clinical presentations of TBDs, particularly of newly emerging viral TBDs in the United States, can help physicians identify the viral TBD, and possibly rule out viral illnesses with different clinical presentations. Therefore, this review discusses clinical manifestations, with an emphasis on dermatologic manifestations of Heartland Virus, Bourbon Virus, Powassan Virus, Deer Tick Virus and Colorado Tick Fever Virus.Viral tick-borne diseases have increased in prevalence over the last decade and often have similar clinical manifestations to other tick-borne diseases, including bacterial infections. Here, we review the dermatologic manifestations of Heartland Virus (HRTV), Bourbon Virus (BRBV), Powassan Virus (POWV), Deer Tick Virus (DTV) and Colorado Tick Fever Virus (CTFV) that are important for clinicians. Tick-borne diseases (TBDs) have doubled in the United States in the last few years, necessitating a greater understanding of the signs and symptoms associated with TBDs for more accurate diagnoses and treatments. . The incBorrelia burgdorferi, spotted fever rickettsiosis (Rickettsia rickettsii and R. parkeri), Tularemia (Francisella tularensis ) and the parasitic agent Babesia microti [Ticks can transmit various pathogens, including bacteria, viruses, and parasites , includi microti . Tick bi microti . In thisPhlebovirus, family Phenuiviridae. HTRV’s genome is a negative single-stranded (-ss) RNA consisting of small, medium, and large segments. The small segment encodes for nucleocapsid protein and nonstructural proteins. The medium segment encodes for structural glycoproteins, Gn and Gc, and are targets of neutralizing antibodies, whereas the large segment encodes RNA-dependent RNA polymerase. Studies have shown that HRTV is genetically related to Dabie bandavirus, formally known as Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV) or Huaiyangshan Banyangvirus [Heartland Virus (HRTV) belongs to the genus angvirus .HRTV was first identified in two Missouri farmers in 2009 . The twoAmblyomma americanum, which was later confirmed as the primary vector of HRTV [Ehrlichia chaffeensis is the bacterial causative agent of HME and is also transmitted by the Lone Star tick in the Midwest region of the United States [Since January 2021, more than 50 cases have been reported in the Midwestern and Southern United States; specifically in Arkansas, Georgia, Illinois, Indiana, Iowa, Kansas, Kentucky, Missouri, North Carolina, Oklahoma, and Tennessee . Most ca of HRTV , 12. Clid States . Symptomd States . Due to Thogotovirus, family Orthomyxoviridae. BRBV is a novel enveloped negative-sense RNA virus and consists of six segments in its genome that are predicted to encode for PB2, PB1, and PA polymerase proteins, a nucleoprotein (NP), a surface glycoprotein (GP), and a matrix (M) protein [Amblyomma americanum) is the vector of BRBV [Bourbon virus (BRBV) belongs to the genus  protein . A singl protein . Like HR of BRBV , 18.BRBV was first identified when a patient from Bourbon County, Kansas, died in 2014 after multiple tick bites. Since the discovery of the Bourbon virus, the virus has been a relatively rare tick-borne viral illness in the United States, with the CDC reporting only a few cases . The casIt has been reported that BRBV-infected individuals develop fever, anorexia, nausea, vomiting, myalgia, and arthralgia. In addition, they may present with diffuse maculopapular or a papular rash that appears on the torso . Blood tSimilarly, the Dhori virus (DHOV) is transmitted by metastriate ticks (non-Ixodes ticks) and mosquitoes. DHOV is found in Europe, North Africa, and western and central Asia and share 70% genome sequence identity with BRBV in multiple genomic segments . Unlike Currently, there are no routine laboratory tests that can confirm the diagnosis of BRBV infections. Only supportive care  can be offered to the BRBV-infected patients since there are currently no vaccines or antiviral treatment approved for BRBV .Flaviviridae (+ ss RNA viruses), genus Flavivirus, that is transmitted by the Ixodes tick species, causing tick-borne encephalitis. The POWV genome comprises seven genes coding for nonstructural proteins and three genes coding for structural proteins, including the capsid and envelope proteins [Powassan virus (POWV) is the only North American member of the family proteins . Recentlproteins . POWV inproteins .The virus was first identified in 1958 and named after a 5-year-old child who died from encephalitis in Powassan, Canada . POWV inPOWV infections can result in neuroinvasive or non-neuroinvasive diseases. Patients with non-neuroinvasive disease present with sore throat, drowsiness, headache, disorientation, and rarely fever. However, POWV infections are primarily neuroinvasive, a distinctive feature compared to other tick-borne illnesses . The neuA faint, diffuse maculopapular rash on the trunk has been described in a few POWV-infected patients , which cDermacentor andersoni, but is mainly found in deer ticks (I. scapularis) collected from the northeastern United States. Three cases of DTV infections have been reported in the literature through 2017 [As mentioned earlier, DTV is a genetic variant (lineage II) of POWV. They share 84% and 94% genomic sequence identity and amino acid sequence identity, respectively, and cannot be distinguished serologically . DTV is ugh 2017 .DTV can cause severe central nervous system infections in humans, similar to POWV. Symptoms include fever, arthralgias, and headache. Laboratory tests of DTV-infected patients show thrombocytopenia with possible acute kidney injury . Unlike Coltivirus, family Reoviridae [Coltivirus have double-stranded RNA genomes made of 12 segments that encode for 13 viral proteins (VP) 1–12 [Dermacentor andersoni ticks. In contrast, the main vertebrate reservoir is Spermophilus lateralis (golden-mantled ground squirrel) and several other mammals, including chipmunks, wild mice, wood rats, wild rabbits, deer, elk, sheep, and coyotes [Colorado Tick Fever Virus (CTFV) is a member of the genus oviridae . VirusesVP) 1–12 . In the  coyotes .CTFV infections are the second most reported arboviral infections in the United States after West Nile virus infections, with approximately 200–400 cases reported annually . CTFV caTypically, CTFV-infected patients present with biphasic fever, headache, myalgia, and fatigue. In addition, laboratory tests show leukopenia and thrombocytopenia. CTFV-infected patients could develop severe complications such as meningitis, encephalitis, and bleeding disorders, and around 20% of patients require hospitalization . HoweverMacular, maculopapular, and petechial rash, together with hyperesthetic skin, can be associated with CTFV infections in 5–15% of cases , 46. A rSimilar to CTFV, Eyach virus (EYAV) which shares 55–88% genetic sequence identity with CTFV presents with a febrile illness . HoweverCTFV infections can be diagnosed using serologic tests to detect anti-coltivirus antibodies. These tests include complement fixation tests, seroneutralization assays, immunofluorescence assay, ELISA, and Western immunoblotting. However, serology may not be timely since antibody production can take 14–21 days. More commonly, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays are used to detect CTFV RNA or the RNA of its cross-reacting serotypes, including CTFV-Ca and Salmon River virus. Intracerebral inoculation of infected human blood into suckling mice can also be used in the isolation and subsequent diagnosis of coltiviruses, though this method is not practical for most laboratories and clinicians. There are no FDA-approved vaccines or antiviral treatment options for CTFV infections, and only supportive care based on the patient’s symptoms is recommended .“One world, one health” is a call for an interdisciplinary approach to science due to the interconnected nature of human actions, animals, and ecological health. The rising number of tick-borne diseases and the newly emerging diseases, such as HRTV and Bourbon virus, is an incentive to manage these diseases with an interdisciplinary approach, especially between practitioners and virologists. This article discusses a multidisciplinary approach by bridging viral knowledge with clinical knowledge to aid clinicians in recognizing presentations of tick-borne diseases.Borrelia burgdorferi, and the petechial rash in Rocky Mountain Spotted Fever, caused by Rickettsia rickettsii [A. americanum, that presents with an annular rash that is almost identical to erythema migrans with lymphocytic dermal infiltrate seen in Lyme disease [Borrelia lonestari, there is some debate regarding its exact etiology [Tick-borne diseases of bacterial origin present with distinctive dermatological manifestations, such as erythema migrans in Lyme disease, caused by ckettsii , 54. Add disease . While Setiology –58.TBDs of bacterial origin are studied more due to the prevalence in the United States, and as stages of the disease with distinct clinical symptoms have been identified. As a result, bacterial tick-borne infections are also stressed in medical schools across the United States and competency exams (USMLE and COMLEX). In contrast, TBDs of viral origin, primarily those recently discovered, have not been studied as extensively, and more research is needed to distinguish characteristic symptoms, like cutaneous presentations. Underreporting and underdiagnosis of viral tick-borne illness can occur due to overlapping dermatological indications of viral and bacterial tick-borne disease. Many viral TBDs can be mistaken for bacterial TBDs without adequate serological testing.Ixodes scapularis tick that carries Lyme disease can also cause co-infections with Powassan virus [Bacterial TBDs and viral TBDs can also often overlap due to ticks harboring multiple diseases. For example, the an virus . Co-infean virus . Co-infeCutaneous presentations of these viral tick-borne illnesses inform practitioners and patients about which viral disease was transmitted from the tick. The knowledge of the viral particle guides treatment and can prevent death. Performing a detailed skin exam on a patient presenting post tick bite or with symptoms after visiting an area endemic with these diseases helps identify cutaneous presentations of illnesses, such as maculopapular rash, erythema migrans, or petechial rash. Further research on the treatment of tick-borne viral diseases is needed to prevent deaths, mainly due to the doubling in tick-borne disease cases in the past ten years in the United States.There are no specific drug therapies, or FDA-approved vaccines for tick-borne viral infections in the United States. While hospitalization is common for these infections, treatment is mainly supportive. Personal protection, landscape management, and wildlife management are effective for preventing and controlling tick-borne viral infectious diseases, but vaccines and drug therapies are critically needed. A convenient area to report a case of tick-borne disease is also required, especially because patients with mild forms of TBDs do not come to the hospital, and those mild symptoms are not documented. A website through the CDC where patients can add their symptoms and their location can aid in solving the underreporting of tick-borne illnesses while giving insights on mild forms of the diseases. The database can also lead to more distinctive dermatological manifestations by stage of the disease Fig. , Table 1


A straightforward electro-conversion of cumene into acetophenone has been reported using boron-doped diamond (BDD) electrodes. This particular conversion is driven by the addition reaction of a cathodically generated hydroperoxide anion to an anodically generated cumyl cation, where the BDD’s wide potential window enables the direct anodic oxidation of cumene into the cumyl cation. Since electricity is directly employed as the oxidizing and reducing reagents, the present protocol is easy to use, suitable for scale-up, and inherently safe. Generally, molecular oxygen has been utilized in the straightforward oxidation of aromatic alkyls. However, since molecular oxygen is highly stable, activation of the molecular oxygen itself is necessary, which requires a specific catalyst and/or harsh conditions such as high temperature and pressure. Recent environmental and sustainable concerns lead to a growing demand for the development of greener oxidation processes. For example, even for the cumene process that involves the oxidation reaction of cumene, first reported in 1944 [Selective oxidation of aromatic alkyl side chains is an important molecular transformation process to obtain various rubbers, resins, fine chemicals, and other industrial products –2: terep in 1944 , a wide  in 1944 .Electro-organic synthesis refers to an organic synthetic method combined with electrochemistry –14. A stHerein, we report the straightforward electro-conversion of cumene, one of the most important and extensively investigated aromatic alkyls, by BDD electrodes. Acetophenone was obtained as the main product when BDD was used as the anode. The role of electrode materials was investigated with electrochemical measurements. Only the BDD anode with a wide potential window can oxidize cumene directly to afford a cumyl cation as the reaction intermediate. Furthermore, acetophenone is produced via cumene hydroperoxide, and this molecular conversion is found to proceed electrochemically.1) in 0.1 M Bu4NBF4/MeCN under constant current conditions in an undivided beaker-type cell (3) and α-cumyl alcohol (4) was also obtained. When the anion of the supporting electrolyte was changed to the perchlorate ion  and the amount of charge  were investigated   , which i1 in MeCN–MeOH to confirm whether the reaction intermediate is a radical or cationic species  electrodes. The BDD’s wide potential window enabled the direct anodic oxidation of cumene to afford a key reaction intermediate, which cannot be realized by other electrodes such as graphite and Ni. Electrosynthesis is a sustainable, scalable, and cost-efficient protocol; a specific catalyst is not required, and reagent waste can be avoided. In addition, the present work offers new perspectives for an electrosynthetic strategy toward oxidation reactions of aromatic alkyls.1, 0.12 g, 1.00 mmol) and supporting electrolyte (0.1 M) in 5 mL solvent was transferred into the electrolysis cell equipped with electrodes . A constant current electrolysis was performed at room temperature. After application of the desired amount of charge, the electrolysis was stopped, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography (CH2Cl2).Electrolysis was carried out by using an IKA screening system . A solution of cumene (File 11H NMR spectra of isolated compounds 2, 3, 4, and 5.Characterization data and


Flaviviridae family includes important viruses, such as Dengue, Zika, West Nile, Japanese encephalitis, Murray Valley encephalitis, tick-borne encephalitis, Yellow fever, Saint Louis encephalitis, and Usutu viruses. They are transmitted by mosquitoes or ticks, and they can infect humans, causing fever, encephalitis, or haemorrhagic fever. The treatment resources for these diseases and the number of vaccines available are limited. It has been discovered that eukaryotic cells synthesize small RNA molecules that can bind specifically to sequences present in messenger RNAs to inhibit the translation process, thus regulating gene expression. These small RNAs have been named microRNAs, and they have an important impact on viral infections. In this review, we compiled the available information on miRNAs that can interact with the 3’ untranslated region (3’UTR) of the flavivirus genome, a conserved region that is important for viral replication and translation.The genus Flavivirus of the  Flaviviridae family are classified into four main genera: Flavivirus, Pegivirus, Pestivirus and Hepacivirus  A, C, and D  or Pestivirus A, bovine viral diarrhoea virus 2 (BVDV-2) or Pestivirus B, classical swine fever virus (CSFV) or Pestivirus C, border disease virus (BDV) or Pestivirus D, and others designated as Pestiviruses E to K (Hepacivirus genus is human hepatitis C virus (HCV), and this genus also includes GBV-B virus (Flavivirus genus includes a large group of viruses that are organized into four groups according to their mechanism of transmission: mosquito-borne flaviviruses (MBFV), transmitted by mosquitoes; tick-borne flaviviruses (TBFV), transmitted by ticks; insect-specific flaviviruses (ISFV), which circulate only among mosquitoes and other insects such as soldier flies and sandflies; and no-known-vector flaviviruses (NKFV), found only in rodents and bats , dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV), Japanese encephalitis virus (JEV), Usutu virus (USUV), Saint Louis encephalitis virus (SLEV), and Murray Valley encephalitis virus (MVEV). TBFV includes tick-borne encephalitis virus (TBEV) (MBFV and TBFV are particularly important for their ability to produce a variety of illnesses in humans. MBFV are transmitted by s (TBEV) .Culex mosquitoes  are small noncoding RNAs (approximately 22-25 nt) that control gene expression at the posttranscriptional level in eukaryotic cells . Two patThen, the miRNA duplex is loaded in the agonist protein (AGO) of the RNA-induced silencing complex (RISC) to promote miRNA-messenger RNA (mRNA) interactions, primarily in the 3´ untranslated region  but also in the 5’UTR and open reading frames (ORFs) of the mRNAs to induce mRNA silencing and decay  or increThe non-canonical miRNA pathway could be processed by several mechanisms in which other noncoding RNAs are involved, including the processing of endogenous introns, snoRNAs, tRNAs, and shRNAs that derivate to unconventional pre-miRNA (approximately 22 nucleotides). It has been reported that non-canonical miRNAs can be produced in a microprocessor-independent or Dicer-independent manner. An example is miRtrons biogenesis, where the pri-miRNAs sequence corresponds to the entire intronic gene, and after the splicing, the pre-miRNA is processed by Dicer to create a mature miRNA . In otheMiRNAs play important roles during flavivirus-host interactions  and it hin vitro , which contains a putative cyclization sequence; and a short hairpin (A2). In DENV, the two dumbbell structures are flanked by A-rich sequences (AR1-3). A five-base motif within AR1 and AR2 can base pair to nucleotides in the loops of DB1 and DB2, leading to the formation of the pseudoknots PK1 and PK2. This basic structure is also observed in JEV, WNV, MVE and KUN. The DB2 structure is absent in YFV and ZIKV, and both DB1 and DB2 are absent in the TBE genome; however, the formation of pseudoknots is apparently a common feature observed in all MBFV . These sThe 3’-terminal regions of MVE, YF and DENV viruses have a cyclization sequence (CS1), a small hairpin (sHP), and a 3’ hairpin structure from 87 to 96 nucleotides .CS1 is a sequence of 26 nucleotides complementary to others present in the 5’ UTR. The interactions between the cyclization sequences in the 5’-and 3’-terminal regions result in a conformational change that is a prerequisite for self-primed viral RNA synthesis by the viral RNA polymerase  and probDownstream of the CS1 sequence, there is a small region with low nucleotide conservation among MBFV. In contrast, in DENV, the CS1 sequence forms a conserved secondary structure designated sHP, which is important for RNA synthesis but not translation. The 3’ UAR, which interacts with the complementary 5’ UAR present in the 5’ UTR, is extended from the sHP to the 3’ hairpin stem .The 3’ hairpin or 3’SL is located at the very end of the viral genome and is also found in NKVF and ISVF . It has The top of the 3’SL structure has been the target for designing synthetic nucleic acid derivatives called peptide nucleic acids (PNA) that inhibit JEV replication in BHK-21 cells . It has In WNV, the 3’ SL structure is essential for viral replication but not for viral translation. It has been demonstrated that the conserved penta-nucleotide 5’ CACAG 3’ located in the top of this stem–loop structure is required for viral replication, particularly the nucleotides in the first, second, third and fifth positions .In the ZIKV genome, at 13 nucleotides from the end of the 3’UTR, a G-quadruplex sequence (PQS) characterized by the presence of two or more contiguous runs of guanosines in a short sequence has been identified. The guanosines fold around potassium cellular ions to form G-tetrads, and they can be used as targets for drugs. These PQSs have been identified in other flaviviruses but within the coding region, not the 3’UTR. These PQSs in RNA molecules have been associated with mRNA splicing, transcriptional termination, and translational control; however, their function in the context of viral genomes is not completely understood. This PQS located at the end of the 3’ end of the ZIKV genome could likely be associated with viral replication .Several studies have identified miRNAs that are targets of cellular genes important to flavivirus infections , but theSome miRNAs have been shown to able to interact with the viral genome with repercussions in the viral replicative cycle Table 1.The antiviral effect shared by several miRNAs has been evaluated in more detail. Many studies have analysed the effects of the insertion of miRNA recognition elements (MREs) into the viral genome Table 2.The above experiments have been useful for studying the influence of miRNAs on flavivirus genomes, but they all employed artificial flaviviruses. Another strategy proposed to control flavivirus infections is the design of miRNAs using the viral genome as a template Table 3.in vitro or in vivo strategies  was prepared in saline buffer. It included hsa-mir-127-3p, hsa-mir-486-5p, hsa-mir-593-5p, and mmu-mir-487b-5p with targets in the 8 genome segments of PR8 (H1N1) influenza virus, and hsa-miR-1-3p with targets in the host ATP6V1A, a protein that regulates viral replication. Intranasal administration of this combination to BALB/c mice resulted in a moderate weight loss and reduction in lung damage after influenza virus infection with reduction in viral mRNA and protein levels . Finallyin vitro and in vivo, and this information will be useful to design antiviral therapies in humans or even strategies directed at the vector to reduce transmission. In this way, miRNAs have a promising future as therapeutic molecules.Flaviviruses are important pathological agents that cause important diseases in humans, from mild fever to encephalitis and haemorrhagic fever, which in many cases are potentially fatal. There are no antiviral drugs to treat diseases caused by flaviviruses, and the number of vaccines to prevent them is still limited. miRNAs are small noncoding RNAs that are able to control eukaryotic gene expression at a posttranscriptional level, but recently, those miRNAs have been discovered to also be capable of interacting specifically with viral genomes and to have either pro- or anti-viral effects. In the case of flaviviruses, almost all miRNas that interact with the viral genome have an inhibitory effect. In fact, this property of some miRNAs has been exploited to design engineered viruses for vaccination purposes; however, although experimental results have been promising, these experiments have only been performed in animal models and with a limited number of viruses. Bioinformatic software is a useful tool that combines a large amount of data and performs predictions that can then be applied to experimental procedures. In fact, the available information on flaviviral genome sequences and miRNAs showed that there are many miRNAs able to interact with the genome of more than one flavivirus, and humans display a higher number than mosquitoes. The miRNAs identified here should be tested RA-B: conceptualization, investigation, and writing. JS-B: conceptualization, investigation, and writing. All authors contributed to the article and approved the submitted version.This work was supported by Secretaria de Investigación y Posgrado of Instituto Politécnico Nacional (Project SIP 20221502). Dr. JS-B has a fellowship from Estímulo al Desempeño de los Investigadores (EDI) and Comisión de Operación y Fomento a las Actividades Académicas (COFAA) of Instituto Politécnico Nacional. Dr. RA-B and JS-B have fellowships from Sistema Nacional de Investigadores of Consejo Nacional de Ciencia y Tecnología (CONACyT), Mexico.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Monkeypox virus can infect several animals, including squirrels, Gambian poached rats, dormice, prairie dogs, monkeys, humans, etc. As reported, about 52 015 laboratory‐confirmed cases, including 18 deaths, have been reported to WHO from 102 member states across all 6 WHO regions from 1 Jan 2022 to 2 Sep 2022. WHO defined the disease as a Public Health Emergency of International Concern (PHEIC) on 21 July 2022. These data showed that monkeypox is novel global threat. Over the past 3 years, the COVID‐19 pandemic has posed a significant global challenge to the public health system. However, other diseases, such as hepatitis of unknown origin in children and monkeypox, have been recently reported in European, African, and American countries. These data showed that monkeypox and other emerging or re‐emerging viruses are novel global threats.Orthopoxvirus genus in the subfamily Chordopoxvirinae of the family Poxviridae. There are 12 species in the Orthopoxvirus genus: Abatino macacapox virus, Akhmeta virus, camelpox virus, cowpox virus, ectromelia virus, monkeypox virus, raccoonpox virus, skunkpox virus, taterapox virus, vaccinia virus, variola virus, and volepox virus. Monkeypox is a zoonotic viral disease, which was first discovered in cynomolgus monkeys in 1958 and then reported in a human infection case in the Democratic Republic of the Congo (DRC) in 1970.https://worldhealthorg.shinyapps.io/mpx_global/). The number of weekly reported new cases globally increased by 3.2% in week 36  compared to the previous week (n = 5026). Furthermore, WHO defines the global risk as moderate globally. However, WHO estimates the risk in the European region as high, and moderate in the African, Americas, Eastern Mediterranean, and Southeast Asia regions. The risk in the Western Pacific region is identified as low‐moderate .Monkeypox is caused by infection with the monkeypox virus (MPXV), a linear, enveloped, double‐stranded DNA (dsDNA) virus belonging to the MPXV can infect several animals, including squirrels, Gambian poached rats, dormice, prairie dogs, monkeys, and humans.The essential elements of the epidemic of infectious diseases include the pathogenic microorganisms , the route of transmission, and the susceptible people or animals. Recently, Bernstein et al. evaluated the cost and benefit of primary prevention of zoonotic infectious diseases.First, for known pathogens, standard procedures of the five or six phases can be taken to control pathogens, cut off transmission routes, and protect susceptible people/animals. However, we cannot wait until the outbreak or severe infectious cases occur before taking appropriate measures. On the contrary, it is necessary to prevent and control the spread of novel pathogens or known pathogens causing severe diseases in advance according to a higher level of prevention and control strategies. For example, the prevention and control strategy adopted by WHO in COVID‐19 was inappropriate. At that time, human‐to‐human and cold‐chain transmission cases of SARS‐CoV‐2 had been reported in China,The second measure is to carry out widespread science propaganda to increase the public's awareness of diseases and their pathogens (including pathogens in the same genus) and improve their awareness of preventing infectious diseases. In the past 30 years, the monkeypox epidemic has spread only in some African countries. However, the recent sharp increase in monkeypox infection cases in non‐endemic countries indicates that the transmission ability of the virus in humans has improved. Monkeypox spreads through close contact, respiratory droplets, and sexual transmission. Although the smallpox vaccine shows high protective efficiency of about 85% against monkeypox, its protective effect will decrease with time. Notably, because smallpox was eradicated in 1980 and the smallpox vaccination program was terminated worldwide, most people have not been vaccinated with the smallpox vaccine. At the same time, the existing smallpox vaccines, modified vaccinia Ankara  and ACAM2000 , are limited, and it is impossible to complete vaccination in a short time. Meanwhile, the clinical symptoms of monkeypox are similar to those of chickenpox and measles, and monkeypox can also be coinfected with herpes simplex virus, HIV, syphilis, or other pathogens. In addition, it will take a lot of work and time to investigate animal sources and transmission routes. Therefore, it is necessary to let the public know about monkeypox and its infectivity and pathogenicity and master the methods to prevent the disease. Widespread science propaganda should be carried out in communities, medical stations, and other places where people live or gather, not just on the websites of WHO and the Centers for Disease Control and Prevention (CDC). In addition, people who contact animals should take necessary precautions, and the hunting and consumption of wild animals should be permanently banned.The third measure is to minimize the spillover of monkeypox from animals, especially rodents and pets. Although monkeypox was first identified in Asian monkeys in Copenhagen in 1958, the MPXV has a broad spectrum of hosts. It is worth noting that except for rodents , monkeys, African hedgehogs, woodchucks, porcupines, giant anteaters, domestic pigs, and nonhuman primates ,The fourth suggestion is that novel diagnostics, vaccines, and therapeutics are still immediately needed. Although several detection methods have been reported and validated,Orthopoxvirus inhibitor in the nonhuman or animal model to explore their exact role in human MPXV infection. Besides, selecting suitable experimental animals is the key to determining the pathogenic mechanism of viruses and evaluating antivirals and vaccines. However, the reliability of replacing experimental animals must be re‐evaluated due to the shortage of experimental monkeys . Furthermore, human or animal health requires the cooperation and support of the whole society to create “One World, One Health.” Therefore, we should work together to control the endemic and build a community with a shared future for humankind.Moreover, although the smallpox vaccine can effectively inhibit monkeypox infection, the live vaccinia virus in the smallpox vaccine can cause rare but life‐threatening side effects, such as encephalitis and progressive vaccinia, especially in immunocompromised people. Therefore, it is important to evaluate the smallpox vaccine and Conceptualization, Linzhu Ren; writing—original draft preparation, Si Chen and Linzhu Ren; writing—review, Linzhu Ren and Liying Zhang; revision, Si Chen and Xue Li; supervision, Linzhu Ren; and funding acquisition, Linzhu Ren. All authors have read and agreed to the published version of the manuscript.The authors declare that they have no competing interests. Linzhu Ren is an Editorial Board member of AMEM and a co‐author of this article. To minimize bias, he was excluded from all editorial decision‐making related to the acceptance of this article for publication.


Infectious evaluation demonstrated monkeypox virus , HSV1 (by PCR) and Streptococcus dysgalactiae (from throat sample); SARS‐CoV‐2 and HSV2 were negative by PCR, as was Epstein‐Barr virus by monospot. Review of peripheral blood (PB) smear demonstrated a normal lymphocyte count (3.55 × 109/L) with scattered highly pleomorphic reactive lymphocytes, from immunoblasts with large nuclei, distinct nucleoli, and abundant basophilic cytoplasm  T‐cell count [501 × 10m Figure  to lymphm Figure . The patThe monkeypox virus belongs to the Poxviridae family, which also includes the molluscum contagiosum and the smallpox; its outbreak has been reported in more than 37 countries since early 2022 . TecovirJared Neeley, Sarah Glogowski, and Weina Chen wrote and revised the manuscript.The authors declare they have no conflicts of interest.The authors received no specific funding for this work.This case followed the institutional patients' ethics. No research on human was performed in this study.