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Plant viruses cause severe losses in agricultural production. Parasite-derived resistance (PDR) offers a promising avenue for developing disease-resistant varieties independent of resistance genes. However, for potyviruses with great agricultural importance, such as soybean mosaic virus (SMV), systematic research on viral genes that can be used for PDR has not been conducted. In this study, we transiently expressed the untranslated region (UTR) or each protein-coding cistron of SMV in Nicotiana benthamiana to evaluate their potential role in conferring PDR. A viral suppressor of RNA silencing (VSR) was also applied to investigate the possible mechanisms of the PDR. The results showed that the transient overexpression of UTR and each cistron of SMV could inhibit SMV infection. The expression of VSR in N. benthamiana leaves could compromise UTR and most of the SMV cistron-mediated inhibition of SMV infection, indicating the involvement of RNA silencing in PDR. In comparison, the expression of VSR could not compromise the PDR conferred by coat protein (CP), P3N-PIPO, cylindrical inclusion (CI), and NIa-Pro, suggesting that these viral cistrons may play roles in PDR at the protein level. These results reveal diverse mechanisms in PDR conferred by different viral cistrons and provide potential gene candidates that can be used for transgenic approaches against SMV. [ABSTRACT FROM AUTHOR]

Soybean mosaic virus (SMV), a member of Potyvirus, is the most destructive and widespread viral disease in soybean production. Our earlier studies identified a soybean 40S ribosomal protein S8 (GmRPS8) using the 6K1 protein of SMV as the bait to screen a soybean cDNA library. The present study aims to identify the interactions between GmRPS8 and SMV and characterize the role of GmRPS8 in SMV infection in soybean. Expression analysis showed higher SMV-induced GmRPS8 expression levels in a susceptible soybean cultivar when compared with a resistant cultivar, suggesting that GmRPS8 was involved in the response to SMV in soybean. Subcellular localization showed that GmRPS8 was localized in the nucleus. Moreover, the yeast two-hybrid (Y2H) experiments showed that GmRPS8 only interacted with 6K1 among the eleven proteins encoded by SMV. The interaction between GmRPS8 and 6K1 was further verified by a bimolecular fluorescence complementation (BiFC) assay, and the interaction was localized in the nucleus. Furthermore, knockdown of GmRPS8 by a virus-induced gene silencing (VIGS) system retarded the growth and development of soybeans and inhibited the accumulation of SMV in soybeans. Together, these results showed that GmRPS8 interacts with 6K1 and contributes to soybean susceptibility to SMV. Our findings provide new insights for understanding the role of GmRPS8 in the SMV infection cycle, which could help reveal potyviral replication mechanisms. [ABSTRACT FROM AUTHOR]

Host proteins are essential during virus infection, and viral factors must target numerous host factors to complete their infectious cycle. The mature 6K1 protein of potyviruses is required for viral replication in plants. However, the interaction between 6K1 and host factors is poorly understood. The present study aims to identify the host interacting proteins of 6K1. Here, the 6K1 of Soybean mosaic virus (SMV) was used as the bait to screen a soybean cDNA library to gain insights about the interaction between 6K1 and host proteins. One hundred and twenty-seven 6K1 interactors were preliminarily identified, and they were classified into six groups, including defense-related, transport-related, metabolism-related, DNA binding, unknown, and membrane-related proteins. Then, thirty-nine proteins were cloned and merged into a prey vector to verify the interaction with 6K1, and thirty-three of these proteins were confirmed to interact with 6K1 by yeast two-hybrid (Y2H) assay. Of the thirty-three proteins, soybean pathogenesis-related protein 4 (GmPR4) and Bax inhibitor 1 (GmBI1) were chosen for further study. Their interactions with 6K1 were also confirmed by bimolecular fluorescence complementation (BiFC) assay. Subcellular localization showed that GmPR4 was localized to the cytoplasm and endoplasmic reticulum (ER), and GmBI1 was located in the ER. Moreover, both GmPR4 and GmBI1 were induced by SMV infection, ethylene and ER stress. The transient overexpression of GmPR4 and GmBI1 reduced SMV accumulation in tobacco, suggesting their involvement in the resistance to SMV. These results would contribute to exploring the mode of action of 6K1 in viral replication and improve our knowledge of the role of PR4 and BI1 in SMV response. [ABSTRACT FROM AUTHOR]

Simple Summary: Wild tomato mosaic virus (WTMV, genus Potyvirus, family Potyviridae) is an emerging viral pathogen that endangers Nicotiana tabacum production. The field survey conducted in this study shows that WTMV is becoming an epidemic in China. An infectious DNA clone of the tobacco-derived WTMV is constructed. It can infect wild eggplant, black nightshade, and tobacco plants but can not infect various local pepper varieties. WTMV evolves into three groups that coincide with their original hosts, tobacco, pepper, or wild eggplant. Thus, the tobacco-derived WTMV might divergently evolves to adapt to tobacco other than peppers. We show that WTMV is compatible with the coinfection of cucumber mosaic virus (CMV) or tobacco mosaic virus (TMV) in tobacco but not other potyviruses. Specifically, WTMV can interfere with the infection of other potyvirus species in tobacco, a phenomenon known as superinfection exclusion previously observed within the same potyviral species. This study contributes essential knowledge on the evolution, infectivity, and recent epidemics of WTMV, and provides the key tool for further disease-resistance and field management studies. Viral diseases of cultivated crops are often caused by virus spillover from wild plants. Tobacco (N. tabacum) is an important economic crop grown globally. The viral pathogens of tobacco are traditional major subjects in virology studies and key considerations in tobacco breeding practices. A positive-strand RNA virus, wild tomato mosaic virus (WTMV), belonging to the genus potyvirus in the family potyviridae was recently found to infect tobacco in China. In this study, diseased tobacco leaf samples were collected in the Henan Province of China during 2020–2021. Several samples from different locations were identified as WTMV positive. An infectious DNA clone was constructed based on one of the WTMV isolates. By using this clone, we found that WTMV from tobacco could establish infections on natural reservoir hosts, demonstrating a possible route of WTMV spillover and overwintering in the tobacco field. Furthermore, the WTMV infection was found to be accompanied by other tobacco viruses in the field. The co-inoculation experiments indicate the superinfection exclusion (SIE) between WTMV and other potyvirus species that infect tobacco. Overall, our work reveals novel aspects of WTMV evolution and infection in tobacco and provides an important tool for further studies of WTMV. [ABSTRACT FROM AUTHOR]

E3-ubiquitin ligases are known to confer abiotic stress responses in plants. In the present study, GmPUB21, a novel U-box E3-ubiquitin ligase-encoding gene, was isolated from soybean and functionally characterized. The expression of GmPUB21, which possesses E3-ubiquitin ligase activity, was found to be significantly up-regulated by drought, salinity, and ABA treatments. The fusion protein GmPUB21-GFP was localized in the cytoplasm, nucleus, and plasma membrane. Transgenic lines of the Nicotiana benthamiana over-expressing GmPUB21 showed more sensitive to osmotic, salinity stress and ABA in seed germination and inhibited mannitol/NaCl-mediated stomatal closure. Moreover, higher reactive oxygen species accumulation was observed in GmPUB21 overexpressing plants after drought and salinity treatment than in wild-type (WT) plants. Contrarily, silencing of GmPUB21 in soybean plants significantly enhanced the tolerance to drought and salinity stresses. Collectively, our results revealed that GmPUB21 negatively regulates the drought and salinity tolerance by increasing the stomatal density and aperture via the ABA signaling pathway. These findings improved our understanding of the role of GmPUB21 under drought and salinity stresses in soybean. [ABSTRACT FROM AUTHOR]

The Ferredoxin-NADP reductase (FNR) gene plays a significant role in NADPH production, carbon assimilation, antioxidation, and cross-talking between chloroplasts and mitochondria in plants. This study aims to know the functional response of the soybean FNR gene (GmFNR) during a soybean mosaic virus (SMV) infection. For this purpose, we developed the bean pod mottle virus (BPMV)-based gene construct (BPMV-GmFNR) and used it to silence the GmFNR gene in resistant and susceptible lines. The results showed that GmFNR expression decreased to 50% in the susceptible line, compared to 40% in the resistant line. The silencing of GmFNR reduces the photosynthetic capacity and CAT activity of both lines compared to their respective controls. In addition, the H2O2 content increased significantly in the susceptible line, whereas the resistant line did not exhibit any change. Further, an SMV infection in the silencing plants of the susceptible line resulted in serious morphological changes and increased the SMV NIa-protease transcript accumulation compared to its control plants. However, the same impact was not observed in the resistant line. The yeast two-hybrid system, BIFC assay, and quantitative real-time polymerase chain reaction (qRT-PCR) analyses revealed that the GmFNR was interacting with EF1A and coincided with the increased SMV accumulation. The results obtained in this study improve the understanding of the soybean FNR gene response during SMV infection and provide a novel insight into the SMV resistance mechanism. [ABSTRACT FROM AUTHOR]