AtGSTU19 and AtGSTU24 as Moderators of the Response of Arabidopsis thaliana to Turnip mosaic virus

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 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

Ultrastructural effects of PVYNTN infection of <em>Capsicum annuum</em> L. cv. Yolo Wonder generative organs; a first step in describing seed transmission

Potato virus Y NTN (PVYNTN), a member of the family Potyviridae, is one of the most important plant viruses. Despite common occurrence of seed transmission process in the Potyviridae, the number or routes of virion entry into seeds are still unclear. Embryos could probably be infected either through host embryogenesis processes or via infection of reproductive tissues, therefore both processes of virus transmission in seeds and pollen grains are likely to be related. Infection by PVY has been studied in detail in host vegetative organs. We investigated, for the first time the impact of infection by the necrotic strain of PVY on Capsicum annuum reproductive organs. We found PVYNTN particles inside C. annuum pollen grains and on the exine surfaces, and PVY epitopes were also found in pollen tubes. We postulate that the male gametophyte in C. annuum could be a source of PVY infection, which may have significance in self-pollinated hosts. We also demonstrated that PVYNTN particles could be detected inside C. annuum seeds on embryo surfaces, while particles and Potyvirus inclusion bodies were observed in endothelium layers. These were mainly detected inside ovarian tissues, that is, in the ovular integuments and nucelli. Changes in both gametophytes strongly indicate that generative organs were a source of PVYNTN infection. Furthermore, we have demonstrated that in C. annuum, PVY was transmitted vertically via seeds

Ultrastructural effects of PVYNTN infection of Capsicum annuum L. cv. Yolo Wonder generative organs; a first step in describing seed transmission

Potato virus Y NTN (PVYNTN), a member of the family Potyviridae, is one of the most important plant viruses. Despite common occurrence of seed transmission process in the Potyviridae, the number or routes of virion entry into seeds are still unclear. Embryos could probably be infected either through host embryogenesis processes or via infection of reproductive tissues, therefore both processes of virus transmission in seeds and pollen grains are likely to be related. Infection by PVY has been studied in detail in host vegetative organs. We investigated, for the first time the impact of infection by the necrotic strain of PVY on Capsicum annuum reproductive organs. We found PVYNTN particles inside C. annuum pollen grains and on the exine surfaces, and PVY epitopes were also found in pollen tubes. We postulate that the male gametophyte in C. annuum could be a source of PVY infection, which may have significance in self-pollinated hosts. We also demonstrated that PVYNTN particles could be detected inside C. annuum seeds on embryo surfaces, while particles and Potyvirus inclusion bodies were observed in endothelium layers. These were mainly detected inside ovarian tissues, that is, in the ovular integuments and nucelli. Changes in both gametophytes strongly indicate that generative organs were a source of PVYNTN infection. Furthermore, we have demonstrated that in C. annuum, PVY was transmitted vertically via seeds

Glutathione-the “master” antioxidant in the regulation of resistant and susceptible host-plant virus-interaction

The interaction between plant hosts and plant viruses is a very unique and complex process, relying on dynamically modulated intercellular redox states and the generation of reactive oxygen species (ROS). Plants strive to precisely control this state during biotic stress, as optimal redox levels enable proper induction of defense mechanisms against plant viruses. One of the crucial elements of ROS regulation and redox state is the production of metabolites, such as glutathione, or the activation of glutathione-associated enzymes. Both of these elements play a role in limiting the degree of potential oxidative damage in plant cells. While the role of glutathione and specific enzymes is well understood in other types of abiotic and biotic stresses, particularly those associated with bacteria or fungi, recent advances in research have highlighted the significance of glutathione modulation and mutations in genes encoding glutathione-associated enzymes in triggering immunity or susceptibility against plant viruses. Apparently, glutathione-associated genes are involved in precisely controlling and protecting host cells from damage caused by ROS during viral infections, playing a crucial role in the host’s response. In this review, we aim to outline the significant improvements made in research on plant viruses and glutathione, specifically in the context of their involvement in susceptible and resistant responses, as well as changes in the localization of glutathione. Analyses of essential glutathione-associated enzymes in susceptible and resistant responses have demonstrated that the levels of enzymatic activity or the absence of specific enzymes can impact the spread of the virus and activate host-induced defense mechanisms. This contributes to the complex network of the plant immune system. Although investigations of glutathione during the plant-virus interplay remain a challenge, the use of novel tools and approaches to explore its role will significantly contribute to our knowledge in the field

Modifications in Tissue and Cell Ultrastructure as Elements of Immunity-Like Reaction in Chenopodium quinoa against Prune Dwarf Virus (PDV)

Prune dwarf virus (PDV) is a plant RNA viral pathogen in many orchard trees worldwide. Our knowledge about resistance genes or resistant reactions of plant hosts to PDV is scant. To fill in part of this gap, an aim of this study was to investigate reactions to PDV infection in a model host, Chenopodium quinoa. Our investigations concentrated on morphological and ultrastructural changes after inoculation with PDV strain 0599. It turned out that PDV infection can cause deformations in host cells but also induce changes in the organelles, such as chloroplasts in inoculated leaves. Moreover, we also demonstrated specific reactions/changes, which could be associated with both types of vascular tissue capable of effectively blocking the systemic spread of PDV to upper leaves. Furthermore, the relative amount of virus, P1 protein deposition, and movement protein (MP) gene expression consequently decreased in PDV-inoculated leaves

Ultrastructural Analysis of Prune Dwarf Virus Intercellular Transport and Pathogenesis

Prune dwarf virus (PDV) is an important viral pathogen of plum, sweet cherry, peach, and many herbaceous test plants. Although PDV has been intensively investigated, mainly in the context of phylogenetic relationship of its genes and proteins, many gaps exist in our knowledge about the mechanism of intercellular transport of this virus. The aim of this work was to investigate alterations in cellular organelles and the cell-to-cell transport of PDV in Cucumis sativus cv. Polan at ultrastructural level. To analyze the role of viral proteins in local transport, double-immunogold assays were applied to localize PDV coat protein (CP) and movement protein (MP). We observe structural changes in chloroplasts, mitochondria, and cellular membranes. We prove that PDV is transported as viral particles via MP-generated tubular structures through plasmodesmata. Moreover, the computer-run 3D modeling reveals structural resemblances between MPs of PDV and of Alfalfa mosaic virus (AMV), implying similarities of transport mechanisms for both viruses

Spatiotemporal Changes in Xylan-1/Xyloglucan and Xyloglucan Xyloglucosyl Transferase (XTH-Xet5) as a Step-In of Ultrastructural Cell Wall Remodelling in Potato–Potato Virus Y (PVYNTN) Hypersensitive and Susceptible Reaction

One type of monitoring system in a plant cell is the cell wall, which intensively changes its structure during interaction with pathogen-stress factors. The wall plays a role as a dynamic and controlled structure, although it is not fully understood how relevant these modifications are to the molecular mechanisms during plant&ndash;virus interactions. In this work we localise the non-cellulosic polysaccharides such as xyloglucan, xylan (xylan-1) and xyloglucosyl transferase (XTH-Xet5), the enzyme that participates in the metabolism of xyloglucan. This provided us with information about the in situ distribution of the components of the hemicellulotic cell wall matrix in hypersensitive and susceptible potato&ndash;PVYNTN interactions. The loosening of the cell wall was accompanied by an increase in xylan depositions during susceptible interactions, whereas, during the hypersensitive response, when the cell wall was reinforced, the xylan content decreased. Moreover, the PVY inoculation significantly redirected XTH-Xet5 depositions, regardless of types of interactions, compared to mock-inoculated tissues. Furthermore, the immunogold localisation clearly revealed the domination of Xet5 in the cell wall and in vesicles in the susceptible host. In contrast, in the resistant host increased levels of Xet5 were observed in cytoplasm, in the cell wall and in the trans-Golgi network. These findings show that the hypersensitive reaction activated XTH-Xet5 in the areas of xyloglucan endo-transglycosylase (XET) synthesis, which was then actively transported to cytoplasm, cell wall and to vacuoles. Our results provide novel insight into cell wall reorganisation during PVYNTN infection as a response to biotic stress factors. These novel findings help us to understand the mechanisms of defence responses that are incorporated into the cell wall signalling network