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Plant resistance, which acts as a selective pressure that affects viral population fitness, leads to the emergence of resistance‐breaking virus strains. Most recessive resistance to potyviruses is related to the mutation of eukaryotic translation initiation factor 4E (eIF4E) or its isoforms that break their interactions with the viral genome‐linked protein (VPg). In this study, we found that the VPg α1–α2 loop, which is essential for binding eIF4E, is the most variable domain of papaya ringspot virus (PRSV) VPg. PRSV VPg with the naturally occurring amino acid substitution of K105Q or E108G in the α1–α2 loop fails to interact with watermelon (Citrullus lanatus) eIF4E but interacts with watermelon eIF(iso)4E instead. Moreover, PRSV carrying these mutations can break the eIF4E‐mediated resistance to PRSV in watermelon accession PI 244019. We further revealed that watermelon eIF(iso)4E with the amino acid substitutions of DNQS to GAAA in the cap‐binding pocket could not interact with PRSV VPg with natural amino acid substitution of K105Q or E108G. Therefore, our finding provides a precise target for engineering watermelon germplasm resistant to resistance‐breaking PRSV isolates. [ABSTRACT FROM AUTHOR]

Tomato cultivars containing the Tm‐22 resistance gene have been widely known to resist tobacco mosaic virus (TMV) and tomato mosaic virus. Tomato brown rugose fruit virus (ToBRFV), a new emerging tobamovirus, can infect tomato plants carrying the Tm‐22 gene. However, the virulence determinant of ToBRFV that overcomes the resistance conferred by the Tm‐22 gene remains unclear. In this study, we substituted the movement protein (MP) encoding sequences between ToBRFV and TMV infectious clones and conducted infectivity assays. The results showed that MP was the virulence determinant for ToBRFV to infect Tm‐22 transgenic Nicotiana benthamiana plants and Tm‐22‐carrying tomato plants. A TMV MP chimera with amino acid residues 60–186 of ToBRFV MP failed to induce hypersensitive cell death in the leaves of Tm‐22 transgenic N. benthamiana plants. Chimeric TMV containing residues 60–186 of ToBRFV MP could, but chimeric ToBRFV containing 61–187 residues of TMV MP failed to infect Tm‐22 transgenic N. benthamiana plants, indicating that 60–186 residues of MP were important for ToBRFV to overcome Tm‐22 gene‐mediated resistance. Further analysis showed that six amino acid residues, H67, N125, K129, A134, I147, and I168 of ToBRFV MP, were critical in overcoming Tm‐22‐mediated resistance in transgenic N. benthamiana plants and tomato plants. These results increase our understanding of the mechanism by which ToBRFV overcomes Tm‐22‐mediated resistance. [ABSTRACT FROM AUTHOR]

Coat proteins (CPs) play critical roles in potyvirus cell‐to‐cell movement. However, the underlying mechanism controlling them remains unclear. Here, we show that substitutions of alanine, glutamic acid, or lysine for the conserved residue tryptophan at position 122 (W122) in tobacco vein banding mosaic virus (TVBMV) CP abolished virus cell‐to‐cell movement in Nicotiana benthamiana plants. In agroinfiltrated N. benthamiana leaf patches, both the CP and RNA accumulation levels of three W122 mutant viruses were significantly reduced compared with those of wild‐type TVBMV, and CP accumulated to a low level similar to that of a replication‐deficient mutant. The results of polyprotein transient expression experiments indicated that CP instability was responsible for the significantly low CP accumulation levels of the three W122 mutant viruses. The substitution of W122 did not affect CP plasmodesmata localization or virus particle formation; however, the substitution significantly reduced the number of virus particles. The wild‐type TVBMV CP could complement the reduced replication and abolished cell‐to‐cell movement of the mutant viruses. When the codon for W122 was mutated to that for a different aromatic residue, phenylalanine or tyrosine, the resultant mutant viruses moved systemically and accumulated up to 80% of the wild‐type TVBMV level. Similar results were obtained for the corresponding amino acids of W122 in the watermelon mosaic virus and potato virus Y CPs. Therefore, we conclude that the aromatic ring in W122 in the core domain of the potyviral CP is critical for cell‐to‐cell movement through the effects on CP stability and viral replication. [ABSTRACT FROM AUTHOR]

Tobacco vein banding mosaic virus (TVBMV) is a potyvirus which mainly infects solanaceous crops. The helper component proteinase (HCpro) of a potyvirus is an RNA silencing suppressor protein and determines the severity of disease symptoms caused by different potyviruses, including TVBMV. It has been shown that substitution mutations introduced into the HCpro open reading frame (ORF) in a TVBMV infectious clone result in changes of Asp189 to Lys or Ile250-Gln251 to Asp-Glu (Asp, aspartic acid; Gln, glutamine; Glu, glutamic acid; Ile, isoleucine). These amino acid changes eliminate the RNA silencing suppression activity of the mutant HCpro (HCm) and attenuate the disease symptoms caused by the mutant TVBMV (T-HCm) in Nicotiana benthamiana plants. Here, we used RNA-sequencing technology to compare gene expression in plants inoculated with the wild-type TVBMV (T-WT) with that in plants inoculated with T-HCm at 1, 2 and 10 days post-agroinfiltration (dpai). At 1 and 2 dpai, N. benthamiana genes related to the translation machinery were up-regulated, whereas genes related to lipid biosynthesis and metabolism or to responses to extracellular/external stimuli were down-regulated in leaves inoculated with T-WT or T-HCm. At 10 dpai, T-WT infection repressed photosynthesis-related genes. T-WT and T-HCm infections differentially perturbed the genes involved in the RNA silencing pathway. The salicylic acid and ethylene signalling pathways were induced, but the jasmonic acid signalling pathway was repressed after T-WT infection. Infections of T-WT and T-HCm also differentially regulated the genes involved in auxin signalling transduction, which is known to associate with the stunting phenotypes caused by TVBMV. These results illustrate the dynamic nature of TVBMV infection in N. benthamiana at the transcriptomic level. [ABSTRACT FROM AUTHOR]

Tomato cultivars containing the Tm-2 2 resistance gene have been widely known to resist tobacco mosaic virus (TMV) and tomato mosaic virus. Tomato brown rugose fruit virus (ToBRFV), a new emerging tobamovirus, can infect tomato plants carrying the Tm-2 2 gene. However, the virulence determinant of ToBRFV that overcomes the resistance conferred by the Tm-2 2 gene remains unclear. In this study, we substituted the movement protein (MP) encoding sequences between ToBRFV and TMV infectious clones and conducted infectivity assays. The results showed that MP was the virulence determinant for ToBRFV to infect Tm-2 2 transgenic Nicotiana benthamiana plants and Tm-2 2 -carrying tomato plants. A TMV MP chimera with amino acid residues 60-186 of ToBRFV MP failed to induce hypersensitive cell death in the leaves of Tm-2 2 transgenic N. benthamiana plants. Chimeric TMV containing residues 60-186 of ToBRFV MP could, but chimeric ToBRFV containing 61-187 residues of TMV MP failed to infect Tm-2 2 transgenic N. benthamiana plants, indicating that 60-186 residues of MP were important for ToBRFV to overcome Tm-2 2 gene-mediated resistance. Further analysis showed that six amino acid residues, H 67 , N 125 , K 129 , A 134 , I 147 , and I 168 of ToBRFV MP, were critical in overcoming Tm-2 2 -mediated resistance in transgenic N. benthamiana plants and tomato plants. These results increase our understanding of the mechanism by which ToBRFV overcomes Tm-2 2 -mediated resistance., (© 2021 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Coat proteins (CPs) play critical roles in potyvirus cell-to-cell movement. However, the underlying mechanism controlling them remains unclear. Here, we show that substitutions of alanine, glutamic acid, or lysine for the conserved residue tryptophan at position 122 (W 122 ) in tobacco vein banding mosaic virus (TVBMV) CP abolished virus cell-to-cell movement in Nicotiana benthamiana plants. In agroinfiltrated N. benthamiana leaf patches, both the CP and RNA accumulation levels of three W 122 mutant viruses were significantly reduced compared with those of wild-type TVBMV, and CP accumulated to a low level similar to that of a replication-deficient mutant. The results of polyprotein transient expression experiments indicated that CP instability was responsible for the significantly low CP accumulation levels of the three W 122 mutant viruses. The substitution of W 122 did not affect CP plasmodesmata localization or virus particle formation; however, the substitution significantly reduced the number of virus particles. The wild-type TVBMV CP could complement the reduced replication and abolished cell-to-cell movement of the mutant viruses. When the codon for W 122 was mutated to that for a different aromatic residue, phenylalanine or tyrosine, the resultant mutant viruses moved systemically and accumulated up to 80% of the wild-type TVBMV level. Similar results were obtained for the corresponding amino acids of W 122 in the watermelon mosaic virus and potato virus Y CPs. Therefore, we conclude that the aromatic ring in W 122 in the core domain of the potyviral CP is critical for cell-to-cell movement through the effects on CP stability and viral replication., (© 2020 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)