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Three gene products of a begomovirus–betasatellite complex restore expression of a transcriptionally silenced green fluorescent protein transgene in Nicotiana benthamiana

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Abstract

Single-stranded DNA geminiviruses replicate via double-stranded DNA intermediates forming mini-chromosomes that are targets for transcriptional gene silencing (TGS) in plants. The ability of the cotton leaf curl Kokhran virus (CLCuKoV)–cotton leaf curl Multan betasatellite (CLCuMuB) proteins, replication-associated protein (Rep), transcriptional activator protein (TrAP), C4, V2 and βC1, to suppress TGS was investigated by using the Nicotiana benthamiana line 16-TGS (16-TGS) harbouring a transcriptionally silenced green fluorescent protein (GFP) transgene. Inoculation of 16-TGS plants with a recombinant potato virus X vector carrying Rep, TrAP or βC1 resulted in re-expression of GFP. Northern blot analysis confirmed that the observed GFP fluorescence was associated with GFP mRNA accumulation. These results indicated that Rep, TrAP and βC1 proteins of CLCuKoV–CLCuMuB can re-activate the expression of a transcriptionally silenced GFP transgene in N. benthamiana. Although Rep, TrAP, or βC1 proteins have, for other begomoviruses or begomoviruses–betasatellites, been previously shown to have TGS suppressor activity, this is the first report demonstrating that a single begomovirus–betasatellite complex encodes three suppressors of TGS.

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Acknowledgments

The authors thank Prof David M. Bisaro (Department of Molecular Genetics, Ohio State University, Columbus, Ohio, USA) for providing the 16-TGS line, Prof. David Baulcombe (Department of Plant Science, University of Cambridge, UK) for providing 16c line, Rob W. Briddon (Plant Biotechnology Division, NIBGE, Faisalabad, Pakistan) for critical reading of the manuscript, Athanasios Dalakouras (RLP AgroScience, Neustadt, Germany) for valuable suggestions and Michèle Zwiebel (RLP AgroScience, Neustadt, Germany) for technical assistance during experiments. This study was funded by the Alexander von Humboldt Foundation, Bonn, Germany.

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Authors and Affiliations

  1. RLP AgroScience GmbH, AlPlanta-Institute for Plant Research, Breitenweg 71, 67435, Neustadt, Germany

    Muhammad Saeed, Gabi Krczal & Michael Wassenegger

  2. Centre for Organismal Studies (COS) Heidelberg, University of Heidelberg, Im Neuenheimer Feld 360, 69120, Heidelberg, Germany

    Michael Wassenegger

  3. National Institute for Biotechnology and Genetic Engineering, Jhang Road, Faisalabad, Pakistan

    Muhammad Saeed

Authors
  1. Muhammad Saeed
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  2. Gabi Krczal
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  3. Michael Wassenegger
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Correspondence to Muhammad Saeed.

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Saeed, M., Krczal, G. & Wassenegger, M. Three gene products of a begomovirus–betasatellite complex restore expression of a transcriptionally silenced green fluorescent protein transgene in Nicotiana benthamiana . Virus Genes 50, 340–344 (2015). https://doi.org/10.1007/s11262-014-1155-8

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  • DOI: https://doi.org/10.1007/s11262-014-1155-8

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