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Recombinant-antibody-mediated resistance against Tomato yellow leaf curl virus in Nicotiana benthamiana

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Abstract

Tomato yellow leaf curl virus (TYLCV) is a geminivirus species whose members cause severe crop losses in the tropics and subtropics. We report the expression of a single-chain variable fragment (scFv) antibody that protected Nicotiana benthamiana plants from a prevalent Iranian isolate of the virus (TYLCV-Ir). Two recombinant antibodies (scFv-ScRep1 and scFv-ScRep2) interacting with the multifunctional replication initiator protein (Rep) were obtained from phage display libraries and expressed in plants, both as stand-alone proteins and as N-terminal GFP fusions. Initial results indicated that both scFvs and both fusions accumulated to a detectable level in the cytosol and nucleus of plant cells. Transgenic plants challenged with TYLCV-Ir showed that the scFv-ScRep1, but more so the fusion proteins, were able to suppress TYLCV-Ir replication. These results show that expression of a scFv-ScRep1-GFP fusion protein can attenuate viral DNA replication and prevent the development of disease symptoms. The present article describes the first successful application of a recombinant antibody-mediated resistance approach against a plant DNA virus.

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Acknowledgments

We would like to thank Dr. Thomas Rademacher (Institute of Molecular Biotechnology, RWTH Aachen University) for his help with fluorescence microscopy and the preparation of plant expression vectors. We acknowledge Prof. Dr. Bruno Gronenborn (Institut des Sciences du Vegetal, CNRS, Gif-sur-Yvette, France) for his useful advice and support while the work was in progress. We thank Dr. Richard Twyman for his critical reading and helpful comments on the preparation of this paper.

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Authors and Affiliations

  1. Institute of Molecular Biotechnology (Biology VII), RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany

    Mohammad Reza Safarnejad, Rainer Fischer & Ulrich Commandeur

  2. Department of Microbial Biotechnology and Biosafety, Agricultural Biotechnology Research Institute, Karaj, Iran

    Mohammad Reza Safarnejad

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  1. Mohammad Reza Safarnejad
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  2. Rainer Fischer
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  3. Ulrich Commandeur
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Correspondence to Ulrich Commandeur.

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Safarnejad, M.R., Fischer, R. & Commandeur, U. Recombinant-antibody-mediated resistance against Tomato yellow leaf curl virus in Nicotiana benthamiana . Arch Virol 154, 457–467 (2009). https://doi.org/10.1007/s00705-009-0330-z

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