Molecular Plant-microbe Interactions最新文献

{"title":"The <i>Fusarium graminearum</i> Effector Protease FgTPP1 Suppresses Immune Responses and Facilitates Fusarium Head Blight Disease.","authors":"Martin Darino, Namrata Jaiswal, Reynaldi Darma, Erika Kroll, Martin Urban, Youhuang Xiang, Moumita Srivastava, Hye-Seon Kim, Ariana Myers, Steven R Scofield, Roger W Innes, Kim E Hammond-Kosack, Matthew Helm","doi":"10.1094/MPMI-08-24-0103-FI","DOIUrl":"10.1094/MPMI-08-24-0103-FI","url":null,"abstract":"<p><p>Most plant pathogens secrete effector proteins to circumvent host immune responses, thereby promoting pathogen virulence. One such pathogen is the fungus <i>Fusarium graminearum</i>, which causes Fusarium head blight (FHB) disease on wheat and barley. Transcriptomic analyses revealed that <i>F. graminearum</i> expresses many candidate effector proteins during early phases of the infection process, some of which are annotated as proteases. However, the contributions of these proteases to virulence remain poorly defined. Here, we characterize an <i>F. graminearum</i> endopeptidase, FgTPP1 (FGSG_11164), that is highly upregulated during wheat spikelet infection and is secreted from fungal cells. To elucidate the potential role of FgTPP1 in <i>F. graminearum</i> virulence, we generated <i>FgTPP1</i> deletion mutants (Δ<i>Fgtpp1</i>) and performed FHB infection assays. Deletion of <i>FgTPP1</i> reduced the virulence of <i>F. graminearum</i> as assessed by spikelet bleaching. Infection with wild-type <i>F. graminearum</i> induced full bleaching in about 50% of the spikes at 10 to 11 days postinfection, whereas this fraction was reduced to between 18 and 27% when using Δ<i>Fgtpp1</i> mutants. Transient expression of green fluorescent protein-tagged FgTPP1 revealed that FgTPP1 localizes, in part, to chloroplasts and attenuates chitin-mediated activation of mitogen-activated protein kinase signaling, reactive oxygen species production, and cell death induced by an autoactive disease resistance protein when expressed in planta. Notably, the FgTPP1 protein is conserved across the Ascomycota phylum, suggesting that it may be a core effector among ascomycete plant pathogens. These properties make FgTPP1 an ideal candidate for decoy substrate engineering, with the goal of engineering resistance to FHB. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"297-314"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143033685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative Analyses of Compatible and Incompatible Host-Pathogen Interactions Provide Insight into Divergent Host Specialization of Closely Related Pathogens.","authors":"Janine Haueisen, Mareike Möller, Heike Seybold, Corinn Small, Mira Wilkens, Lovis Jahneke, Leonhard Parchinger, Elisha Thynne, Eva H Stukenbrock","doi":"10.1094/MPMI-10-24-0133-FI","DOIUrl":"10.1094/MPMI-10-24-0133-FI","url":null,"abstract":"<p><p>Host-pathogen co-evolutionary dynamics drive constant changes in plant pathogens to thrive in their plant host. Factors that determine host specificity are diverse and range from molecular and morphological strategies to metabolic and reproductive adaptations. We applied an experimental approach and conducted comparative microscopy, transcriptome analyses, and functional analyses of selected pathogen traits to identify determinants of host specificity in an important wheat pathogen. We included three closely related fungal pathogens, <i>Zymoseptoria tritici</i>, <i>Z</i>. <i>pseudotritici</i>, and <i>Z</i>. <i>ardabiliae</i>, that establish compatible and incompatible interactions with wheat. Although infections of the incompatible species induce plant defenses during invasion of stomatal openings, we found a conserved early-infection program among the three species whereby only 9.2% of the 8,885 orthologous genes are significantly differentially expressed during initial infection. The genes upregulated in <i>Z. tritici</i> likely reflect specialization to wheat, whereas upregulated genes in the incompatible interaction may reflect processes to counteract cellular stress associated with plant defenses. We selected nine candidate genes encoding putative effectors and host-specificity determinants in <i>Z. tritici</i> and deleted these to study their functional relevance. Despite the particular expression patterns of the nine genes, only two mutants were impaired in virulence. We further expressed the <i>Z. tritici</i> proteins in <i>Nicotiana benthamiana</i> to investigate protein function and assess cell death reaction. Hereby, we identify three effectors with cell-death-inducing properties. From the functional analyses, we conclude that the successful infection of <i>Z. tritici</i> in wheat relies on an extensive redundancy of virulence determinants. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"235-251"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143502432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Triple Threat: How Global Fungal Rice and Wheat Pathogens Utilize Comparable Pathogenicity Mechanisms to Drive Host Colonization.","authors":"Rachel E Kalicharan, Jessie Fernandez","doi":"10.1094/MPMI-09-24-0106-FI","DOIUrl":"10.1094/MPMI-09-24-0106-FI","url":null,"abstract":"<p><p>Plant pathogens pose significant threats to global cereal crop production, particularly for essential crops such as rice and wheat, which are fundamental to global food security and provide nearly 40% of the global caloric intake. As the global population continues to rise, increasing agricultural production to meet food demands becomes even more critical. However, the production of these vital crops is constantly threatened by phytopathological diseases, especially those caused by fungal pathogens such as <i>Magnaporthe oryzae</i>, the causative agent of rice blast disease; <i>Fusarium graminearum</i>, responsible for <i>Fusarium</i> head blight in wheat; and <i>Zymoseptoria tritici</i>, the source of Septoria tritici blotch. All three pathogens are hemibiotrophic, initially colonizing the host through a biotrophic, symptomless lifestyle, followed by causing cell death through the necrotrophic phase. Additionally, they deploy a diverse range of effectors, including proteinaceous and non-proteinaceous molecules, to manipulate fundamental host cellular processes, evade immune responses, and promote disease progression. This review discusses recent advances in understanding the effector biology of these three pathogens, highlighting both the shared functionalities and unique molecular mechanisms they employ to regulate conserved elements of host pathways, such as directly manipulating gene transcription in host nuclei, disrupting reactive oxygen species signaling, interfering with protein stability, and undermining host structural integrity. By detailing these complex interactions, the review explores potential targets for innovative control measures and emphasizes the need for further research to develop effective strategies against these destructive pathogens in the face of evolving environmental and agricultural challenges. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"173-186"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142979211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic Gene-for-Gene Interactions Undermine Durable Resistance.","authors":"Barbara Valent","doi":"10.1094/MPMI-02-25-0022-HH","DOIUrl":"https://doi.org/10.1094/MPMI-02-25-0022-HH","url":null,"abstract":"<p><p>Harold Flor's gene-for-gene model explained boom-bust cycles in which resistance (<i>R</i>) genes are deployed in farmers' fields, only to have pathogens overcome resistance by modifying or losing corresponding active avirulence (<i>AVR</i>) genes. Flor understood that host <i>R</i> genes with corresponding low rates of virulence mutation in the pathogen should maintain resistance for longer periods of time. This review focuses on <i>AVR</i> gene dynamics of the haploid Ascomycete fungus <i>Pyricularia oryzae</i>, which causes rice blast disease, a gene-for-gene system with a complex race structure and a very rapid boom-bust cycle due to high rates of <i>AVR</i> gene mutation. Highly mutable blast <i>AVR</i> genes are often characterized by deletion and by movement to new chromosomal locations, implying a loss/regain mechanism in response to <i>R</i> gene deployment. Beyond rice blast, the recent emergence of two serious new blast diseases on wheat and <i>Lolium</i> ryegrasses highlighted the role of <i>AVR</i> genes that act at the host genus level and serve as infection barriers that separate host genus-specialized <i>P. oryzae</i> subpopulations. Wheat and ryegrass blast diseases apparently evolved through sexual crosses involving fungal individuals from five host-adapted subpopulations, with the host jump enabled by the introduction of virulence alleles of key host-specificity <i>AVR</i> genes. Despite identification of wheat <i>AVR</i>/<i>R</i> gene interactions operating at the host genus specificity level, the paucity of effective <i>R</i> genes identified thus far limits control of wheat blast disease. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":"38 2","pages":"104-117"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plant Viral Synergism: Co-expression of P1 and NIaPro Cistrons of Wheat Streak Mosaic Virus and Triticum Mosaic Virus Is Required for Synergistic Interaction in Wheat.","authors":"Chi Hzeng Wong, Jeffrey Alexander, Satyanarayana Tatineni","doi":"10.1094/MPMI-10-24-0126-FI","DOIUrl":"10.1094/MPMI-10-24-0126-FI","url":null,"abstract":"<p><p>Synergistic interactions among unrelated viruses in mixed infections can cause significant yield losses, and viral determinants of these interactions are poorly understood. Wheat (<i>Triticum aestivum</i> L.) co-infection with wheat curl mite-transmitted wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) results in disease synergism with a drastically increased symptom phenotype of stunted growth, leaf bleaching, and enhanced titers of both viruses compared with individual virus infections. In this study, we examined the viral determinants responsible for WSMV-TriMV disease synergism through transient expression of select cistrons of WSMV in wheat through TriMV and vice-versa. We found that expression of WSMV P1, NIa, or NIaPro in wheat through TriMV or vice-versa elicited moderate to severe symptoms with a moderate or no increase in virus titer. However, co-expression of P1 and NIaPro of WSMV in wheat through TriMV or vice-versa exhibited a WSMV-TriMV disease synergism-like phenotype with enhanced accumulation of genomic RNA copies and coat protein. Additionally, we found that the P3 of both viruses is dispensable for synergism. HCPro and NIaVPg of WSMV and TriMV are not the primary determinants but might have a minor role in efficient synergism. In co-infected wheat, the accumulation of virus-specific small interfering RNAs (vsiRNAs) was increased, similar to viral genomic RNA copies, despite the presence of two viral RNA-silencing suppressors (VRSS), which function through sequestration of vsiRNAs. Our findings revealed that WSMV-TriMV disease synergism is not caused by the suppression of host posttranscriptional gene silencing by two VRSS proteins in co-infected wheat, and the P1 and NIaPro of both viruses collectively drive synergistic interactions between WSMV and TriMV in wheat. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 \"No Rights Reserved\" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2025.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"328-343"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142624222","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protein Kinase-Major Sperm Protein (PK-MSP) Genes Mediate Recognition of the Fungal Necrotrophic Effector SnTox3 to Cause Septoria nodorum Blotch in Wheat.","authors":"Zengcui Zhang, Katherine L D Running, Sudeshi Seneviratne, Amanda R Peters Haugrud, Agnes Szabo-Hever, Gurminder Singh, Kateřina Holušová, István Molnár, Jaroslav Doležel, Timothy L Friesen, Justin D Faris","doi":"10.1094/MPMI-10-24-0125-FI","DOIUrl":"10.1094/MPMI-10-24-0125-FI","url":null,"abstract":"<p><p>The wheat-<i>Parastagonospora nodorum</i> pathosystem has emerged as a model system for plant-necrotrophic fungal pathogen interactions. In this system, fungal necrotrophic effectors are recognized by specific host genes in an inverse gene-for-gene manner to induce programmed cell death and other host responses, which leads to disease. We previously cloned a wheat gene (<i>Snn3-D1</i>) encoding protein kinase and major sperm protein domains that recognizes the <i>P. nodorum</i> necrotrophic effector SnTox3. Here, we identified an <i>Snn3-D1</i> homoeolog (<i>Snn3-B1</i>) and a paralog (<i>Snn3-B2</i>) that also recognize SnTox3, leading to susceptibility. DNA sequence divergence of <i>Snn3-B1</i> and <i>Snn3-B2</i> and differences in transcriptional expression patterns and three-dimensional protein conformation were associated with a more severe programmed cell death response conferred by <i>Snn3-B2</i> compared with <i>Snn3-B1</i>. Both Snn3 proteins were localized to the nucleus and cytoplasm in wheat protoplasts, suggesting that they may have acquired novel functions compared with previously characterized major sperm protein domain-containing proteins in other species. <i>Snn3-B2</i> was previously shown to govern osmotic stress and salt tolerance, indicating that protein kinase-major sperm protein genes can act in plant defense responses to both biotic and abiotic stresses. Evaluation of a large collection of wheat lines showed that several alleles of each gene, including absent alleles, exist within the germplasm. Diagnostic markers were developed for the absent alleles of both genes, which will prove useful for marker-assisted selection in wheat to eliminate SnTox3 sensitivity and achieve better disease resistance. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 \"No Rights Reserved\" license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2025.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"315-327"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143753672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Technical Advances Drive the Molecular Understanding of Effectors from Wheat and Barley Powdery Mildew Fungi.","authors":"Merle Bilstein-Schloemer, Marion C Müller, Isabel M L Saur","doi":"10.1094/MPMI-12-24-0155-FI","DOIUrl":"10.1094/MPMI-12-24-0155-FI","url":null,"abstract":"<p><p>Pathogens manipulate host physiology through the secretion of virulence factors (effectors) to invade and proliferate on the host. The molecular functions of effectors inside plant hosts have been of interest in the field of molecular plant-microbe interactions. Obligate biotrophic pathogens, such as rusts and powdery mildews, cannot proliferate outside of plant hosts. In addition to the inhibition of the plant's immune components, these pathogens are under particular pressure to extract nutrients efficiently from the host. Understanding the molecular basis of infections mediated by obligate biotrophic pathogens is significant because of their impact in modern agriculture. In addition, powdery mildews serve as excellent models for obligate biotrophic cereal pathogens. Here, we summarize the current knowledge on the effectorome of the barley and wheat powdery mildews and putative molecular virulence functions of effectors. We emphasize the availability of comprehensive genomic, transcriptomic, and proteomic resources and discuss the methodological approaches used for identifying candidate effectors, assessing effector virulence traits, and identifying effector targets in the host. We highlight established and more recently employed methodologies, discuss limitations, and suggest additional strategies. We identify open questions and discuss how addressing them with currently available resources will enhance our understanding of <i>Blumeria</i> candidates for secretor effector proteins. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"213-225"},"PeriodicalIF":3.2,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142971635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulation and Functions of Long Noncoding RNAs During <i>Meloidogyne incognita</i> Parasitism of Tomato.","authors":"Selin Ozdemir, Sobhan Bahrami Zadegan, Mst Shamira Sultana, Nicole Coffey, J Hollis Rice, Tarek Hewezi","doi":"10.1094/MPMI-10-24-0140-R","DOIUrl":"10.1094/MPMI-10-24-0140-R","url":null,"abstract":"<p><p>Long noncoding RNAs (lncRNAs) are emerging as important regulators of various aspects of immune response and plant-pathogen interactions. However, the regulatory function of lncRNAs during plant-nematode interaction remains largely elusive. In this study, we investigated the differential regulation and function of lncRNAs during two different stages of tomato infection by the root-knot nematode <i>Meloidogyne incognita</i>. At the early stage of infection, 2,218 and 2,827 lncRNAs were regulated locally in the <i>M. incognita</i>-induced galls and systemically in the neighboring root cells, respectively. However, at the later stage of infection, the number of <i>M. incognita</i>-regulated lncRNAs was dramatically reduced, with only 49 lncRNAs being identified as differentially expressed. Differentially expressed lncRNAs were predicted to encode peptides with functionally annotated domains, providing insights into the potential roles of these peptides in regulating gene expression, RNA stability and splicing, and protein-protein-interactions. Among the differentially expressed lncRNAs, 55 were found to contain putative binding sites for 56 microRNAs (miRNAs). Overexpressing five of these lncRNAs significantly increased tomato resistance to <i>M. incognita</i>, supporting the functional importance of lncRNAs for establishing tomato-<i>M. incognita</i> interaction. Functional analysis of the target mimicry of lncRNAs towards miRNAs resulted in the identification of two novel regulatory modules involving miR47 and miR156e-5p and their targeted genes that regulate tomato responses to <i>M. incognita</i> parasitism. Taken together, our data provide novel insights into the transcriptional and posttranscriptional regulatory functions of lncRNA and open a new avenue to engineer crop plants with enhanced nematode resistance by leveraging the regulatory potential of lncRNAs. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"72-83"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142676464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Michael Mishkind: MPMI Community Builder.","authors":"Jeff Dangl, Guo-Liang Wang, Brad Day, Nicole Donofrio","doi":"10.1094/MPMI-01-25-0011-LE","DOIUrl":"10.1094/MPMI-01-25-0011-LE","url":null,"abstract":"","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"6-8"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143492992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Visualizing Tomato Spotted Wilt Virus Protein Localization: Cross-Kingdom Comparisons of Protein-Protein Interactions.","authors":"K M Martin, Y Chen, M A Mayfield, M Montero-Astúa, A E Whitfield","doi":"10.1094/MPMI-09-24-0108-R","DOIUrl":"10.1094/MPMI-09-24-0108-R","url":null,"abstract":"<p><p>Tomato spotted wilt virus (TSWV) is an orthotospovirus that infects both plants and insect vectors, and understanding its protein localization and interactions is crucial for unraveling the infection cycle and host-virus interactions. We investigated and compared the localization of TSWV proteins. The localization between plant and insect cells was overall consistent, indicating a similar mechanism is utilized by the virus in both types of cells. However, a change in localization over time was associated with the viral proteins that did not contain signal peptides and transmembrane domains such as N, NSs, and NSm, which only occurred in the plant cells and not in the insect cells. We also tested the localization of the proteins during an active plant infection using free red fluorescent protein (RFP) as a marker to highlight the nucleus and cytoplasm. Voids in the cytoplasm were shown only during infection, and N, NSs, NSm, and to a lesser extent G_N and G_C were surrounding these areas, suggesting it may be a site of replication or morphogenesis. Furthermore, we tested the interactions of viral proteins using both bimolecular fluorescence complementation (BiFC) and membrane-based yeast two-hybrid (MbY2H) assays. These revealed self-interactions of NSm, N, G_N, G_C, and NSs. We also identified interactions between different TSWV proteins, indicating their possible roles, such as between NSs and G_C and N and G_C, which may be necessary during the replication and assembly processes, respectively. This research expands our knowledge of TSWV infection and elaborates on the intricate relationships between viral proteins, cellular dynamics, and host responses. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.</p>","PeriodicalId":19009,"journal":{"name":"Molecular Plant-microbe Interactions","volume":" ","pages":"84-96"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142470489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}