Turnip crinkle virus with nonviral gene cancels the effect of silencing suppressors of P19 and 2b in Arabidopsis thaliana |
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| Institution: | 1. Laboratory of Combustion and Green Technologies, Department of Energetic Engineering, University Institute of Technology, University of Ngaoundere, P.O. Box 455, Cameroon;2. Department of Physics, Faculty of Sciences, University of Yaounde 1, Cameroon;3. Institut Pprime, UPR 3346 du CNRS, ENSMA, Poitiers, France;1. Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi 110012, India;2. Division of Biochemistry, Indian Agricultural Research Institute, New Delhi 110012, India;3. Department of Botany & Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia;1. Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA;2. Argonne National Laboratory, Lemont, IL, USA;3. Data Science and Information Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA;1. National Plant Protection Organization, National Reference Centre, P.O. Box 9102, 6700 HC Wageningen, The Netherlands;2. Naktuinbouw, P.O. Box 40, 2370 AA Roelofarendsveen, The Netherlands;3. Instituto de Biología Molecular y Celular de Plantas (UPV-CSIC), Universidad Politécnica de Valencia, Valencia 46022, Spain |
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| Abstract: | In plants, green fluorescent protein (GFP) has become a preferred molecular marker for gene expression and cellular localization, and plant viral vectors are valuable tools for heterologous gene expression. Some plant viruses have been used for expression of GFP, and the activities of these viruses are barely affected by the extra GFP gene. In contrast, the packaging and the length of Turnip crinkle virus (TCV) genome is strictly limited when foreign genes are inserted into the coding sequences of TCV genome. In this report, we removed the silencing suppressor p38 from TCV, and constructed GFP derivatives of TCV. Then the resulting TCV mutants were used to infect Arabidopsis plants containing mutations in key silencing pathway genes, including triple dcl2/dcl3/dcl4, dcl2, dcl4 and ago mutant plants. Our results demonstrate that the activity of TCV is affected by nonviral GFP insert in Arabidopsis plants, and RNA silencing appears not play an important role. AGOs appear to be more efficient at slicing RNAs of viral origin, especially AGO2 and AGO7. Although the viral suppressors of RNA silencing (VSRs) P19 and 2b can enhance the accumulation of viral RNAs, neither P19 nor 2b can significantly increase the expression of TCV mutants with nonviral genes. TCV is an example of an RNA virus that is recalcitrant to add nonviral gene sequences. |
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| Keywords: | Nonviral gene Silencing suppressors Silencing pathway genes RNA silencing |
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