Members of the ribosomal protein S6 (RPS6) family act as pro-viral factor for tomato spotted wilt orthotospovirus infectivity in Nicotiana benthamiana |
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Authors: | Tieme A Helderman Laurens Deurhof André Bertran Manon M S Richard Richard Kormelink Marcel Prins Matthieu H A J Joosten Harrold A van den Burg |
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Institution: | 1. Molecular Plant Pathology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, Netherlands;2. Laboratory of Phytopathology, Department of Plant Sciences, Wageningen University, Wageningen, Netherlands;3. Laboratory of Virology, Department of Plant Sciences, Wageningen University, Wageningen, Netherlands;4. Molecular Plant Pathology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, Netherlands
KeyGene N.V., Wageningen, Netherlands |
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Abstract: | To identify host factors for tomato spotted wilt orthotospovirus (TSWV), a virus-induced gene silencing (VIGS) screen using tobacco rattle virus (TRV) was performed on Nicotiana benthamiana for TSWV susceptibility. To rule out any negative effect on the plants’ performance due to a double viral infection, the method was optimized to allow screening of hundreds of clones in a standardized fashion. To normalize the results obtained in and between experiments, a set of controls was developed to evaluate in a consist manner both VIGS efficacy and the level of TSWV resistance. Using this method, 4532 random clones of an N. benthamiana cDNA library were tested, resulting in five TRV clones that provided nearly complete resistance against TSWV. Here we report on one of these clones, of which the insert targets a small gene family coding for the ribosomal protein S6 (RPS6) that is part of the 40S ribosomal subunit. This RPS6 family is represented by three gene clades in the genome of Solanaceae family members, which were jointly important for TSWV susceptibility. Interestingly, RPS6 is a known host factor implicated in the replication of different plant RNA viruses, including the negative-stranded TSWV and the positive-stranded potato virus X. |
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Keywords: | disease susceptibility forward genetics genetic screen tomato spotted wilt virus Tospoviridae viral susceptibility factors virus-induced gene silencing |
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