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      The ORF2 protein of Fusarium graminearum virus 1 suppresses the transcription of FgDICER2 and FgAGO1 to limit host antiviral defences

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          Summary

          The filamentous fungus Fusarium graminearum possesses an RNA‐interference (RNAi) pathway that acts as a defence response against virus infections and exogenous double‐stranded (ds) RNA. Fusarium graminearum virus 1 (FgV1), which infects F. graminearum, confers hypovirulence‐associated traits such as reduced mycelial growth, increased pigmentation and reduced pathogenicity. In this study, we found that FgV1 can suppress RNA silencing by interfering with the induction of FgDICER2 and FgAGO1, which are involved in RNAi antiviral defence and the hairpin RNA/RNAi pathway in F. graminearum. In an FgAGO1‐ or FgDICER2‐promoter/GFP‐reporter expression assay the green fluorescent protein (GFP) transcript levels were reduced in FgV1‐infected transformed mutant strains. By comparing transcription levels of FgDICER2 and FgAGO1 in fungal transformed mutants expressing each open reading frame (ORF) of FgV1 with or without a hairpin RNA construct, we determined that reduction of FgDICER2 and FgAGO1 transcript levels requires only the FgV1 ORF2‐encoded protein (pORF2). Moreover, we confirmed that the pORF2 binds to the upstream region of FgDICERs and FgAGOs in vitro. These combined results indicate that the pORF2 of FgV1 counteracts the RNAi defence response of F. graminearum by interfering with the induction of FgDICER2 and FgAGO1 in a promoter‐dependent manner.

          Abstract

          The ORF2 protein of Fusarium graminearum virus 1 can directly repress the induction of FgDICER2 and FgAGO1 at the transcriptional level to counteract the RNAi defence response of Fusarium graminearum.

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          Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interactions.

          The gel electrophoresis mobility shift assay (EMSA) is used to detect protein complexes with nucleic acids. It is the core technology underlying a wide range of qualitative and quantitative analyses for the characterization of interacting systems. In the classical assay, solutions of protein and nucleic acid are combined and the resulting mixtures are subjected to electrophoresis under native conditions through polyacrylamide or agarose gel. After electrophoresis, the distribution of species containing nucleic acid is determined, usually by autoradiography of 32P-labeled nucleic acid. In general, protein-nucleic acid complexes migrate more slowly than the corresponding free nucleic acid. In this protocol, we identify the most important factors that determine the stabilities and electrophoretic mobilities of complexes under assay conditions. A representative protocol is provided and commonly used variants are discussed. Expected outcomes are briefly described. References to extensions of the method and a troubleshooting guide are provided.
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            viral silencing suppressors: Tools forged to fine-tune host-pathogen coexistence.

            RNA silencing is a homology-dependent gene inactivation mechanism that regulates a wide range of biological processes including antiviral defense. To deal with host antiviral responses viruses evolved mechanisms to avoid or counteract this, most notably through expression of viral suppressors of RNA silencing. Besides working as silencing suppressors, these proteins may also fulfill other functions during infection. In many cases the interplay between the suppressor function and other "unrelated" functions remains elusive. We will present host factors implicated in antiviral pathways and summarize the current status of knowledge about the diverse viral suppressors' strategies acting at various steps of antiviral silencing in plants. Besides, we will consider the multi-functionality of these versatile proteins and related biochemical processes in which they may be involved in fine-tuning the plant-virus interaction. Finally, we will present the current applications and discuss perspectives of the use of these proteins in molecular biology and biotechnology.
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              Facilitative and antagonistic interactions between plant viruses in mixed infections.

              Mixed infections of plant viruses are common in nature, and a number of important virus diseases of plants are the outcomes of interactions between causative agents. Multiple infections lead to a variety of intrahost virus-virus interactions, many of which may result in the generation of variants showing novel genetic features, and thus change the genetic structure of the viral population. Hence, virus-virus interactions in plants may be of crucial significance for the understanding of viral pathogenesis and evolution, and consequently for the development of efficient and stable control strategies. The interactions between plant viruses in mixed infections are generally categorized as synergistic or antagonistic. Moreover, mixtures of synergistic and antagonistic interactions, creating usually unpredictable biological and epidemiological consequences, are likely to occur in plants. The mechanisms of some of these are still unknown. This review aims to bring together the current knowledge on the most commonly occurring facilitative and antagonistic interactions between related or unrelated viruses infecting the same host plant. The best characterized implications of these interactions for virus-vector-host relationships are included. The terms 'synergism' and 'helper dependence' for facilitative virus-virus interactions, and 'cross-protection' and 'mutual exclusion' for antagonistic interactions, are applied in this article. © 2011 THE AUTHOR. MOLECULAR PLANT PATHOLOGY © 2011 BSPP AND BLACKWELL PUBLISHING LTD.
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                Author and article information

                Contributors
                kookkim@snu.ac.kr
                Journal
                Mol Plant Pathol
                Mol. Plant Pathol
                10.1111/(ISSN)1364-3703
                MPP
                Molecular Plant Pathology
                John Wiley and Sons Inc. (Hoboken )
                1464-6722
                1364-3703
                09 December 2019
                February 2020
                : 21
                : 2 ( doiID: 10.1111/mpp.v21.2 )
                : 230-243
                Affiliations
                [ 1 ] Plant Genomics and Breeding Institute Seoul National University Seoul Korea
                [ 2 ] Department of Agricultural Biotechnology Seoul National University Seoul Korea
                [ 3 ] Research Institute of Agriculture and Life Sciences Seoul National University Seoul Korea
                Author notes
                [*] [* ] Correspondence: Email: kookkim@ 123456snu.ac.kr

                Author information
                https://orcid.org/0000-0001-9066-6903
                Article
                MPP12895
                10.1111/mpp.12895
                6988435
                31815356
                3e272d1d-f675-4dc5-b57d-ecef8a493910
                © 2019 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                Page count
                Figures: 6, Tables: 1, Pages: 14, Words: 17952
                Funding
                Funded by: Ministry of Agriculture, Food and Rural Affairs , open-funder-registry 10.13039/501100003624;
                Award ID: 710011‐03
                Funded by: National Research Foundation , open-funder-registry 10.13039/501100001321;
                Award ID: NRF‐2016R1D1A1B03936370
                Funded by: Ministry of Education , open-funder-registry 10.13039/501100002701;
                Award ID: 710011‐03
                Funded by: Ministry of Agriculture , open-funder-registry 10.13039/100008921;
                Award ID: 21
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                February 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.5 mode:remove_FC converted:29.01.2020

                Plant science & Botany
                fusarium graminearum virus 1,mycovirus,orf2,rnai,silencing suppressor
                Plant science & Botany
                fusarium graminearum virus 1, mycovirus, orf2, rnai, silencing suppressor

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