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Camelid nanobodies with high affinity for broad bean mottle virus: a possible promising tool to immunomodulate plant resistance against viruses

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Abstract

Worldwide, plant viral infections decrease seriously the crop production yield, boosting the demand to develop new strategies to control viral diseases. One of these strategies to prevent viral infections, based on the immunomodulation faces many problems related to the ectopic expression of specific antibodies in planta. Camelid nanobodies, expressed in plants, may offer a solution as they are an attractive tool to bind efficiently to viral epitopes, cryptic or not accessible to conventional antibodies. Here, we report a novel, generic approach that might lead to virus resistance based on the expression of camelid specific nanobodies against Broad bean mottle virus (BBMV). Eight nanobodies, recognizing BBMV with high specificity and affinity, were retrieved after phage display from a large ‘immune’ library constructed from an immunized Arabic camel. By an in vitro assay we demonstrate how three nanobodies attenuate the BBMV spreading in inoculated Vicia faba plants. Furthermore, the in planta transient expression of these three selected nanobodies confirms their virus neutralizing capacity. In conclusion, this report supports that plant resistance against viral infections can be achieved by the in vivo expression of camelid nanobodies.

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Acknowledgments

The authors would like to thank the Director General of the Atomic Energy Commission of Syria and the head of the Molecular Biology and Biotechnology department for their continuous support throughout this work. We thank Mr. Adnan Al-Assad at General Commission for Scientific Agricultural Research (GCSAR), Deir El-Hagar Station for Camelid Research (Damascus, Syria), for housing and immunizing camels. We also thank the lab. of Patrice Dunoyer, IBMP/CNRS institute, Strasbourg, FRANCE for kindly providing pBI61-P19 construct and the lab of Sunil K. Mukherjee, ICGEB, New-Delhi, INDIA for kindly providing empty pBI121 vector.

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Authors and Affiliations

  1. Division of Plant Pathology, Department of Molecular Biology and Biotechnology, Atomic Energy Commission of Syria (AECS), P. O. Box 6091, Damascus, Syria

    Ahmed Ghannam

  2. Plant Virology lab., International Center for Agricultural Research in the Dry Areas (ICARDA), P. O. Box 114/5055, Beirut, Lebanon

    Safa Kumari

  3. Laboratorium voor Cellulaire en Moleculaire Immunologie, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussel, Belgium

    Serge Muyldermans

  4. Structural Biology Research Center, VIB, Brussel, Belgium

    Serge Muyldermans

  5. Division of Microbiology and Immunology, Department of Molecular Biology and Biotechnology, AECS, P. O. Box 6091, Damascus, Syria

    Abdul Qader Abbady

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  1. Ahmed Ghannam
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  2. Safa Kumari
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  3. Serge Muyldermans
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  4. Abdul Qader Abbady
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Correspondence to Ahmed Ghannam or Abdul Qader Abbady.

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Ghannam, A., Kumari, S., Muyldermans, S. et al. Camelid nanobodies with high affinity for broad bean mottle virus: a possible promising tool to immunomodulate plant resistance against viruses. Plant Mol Biol 87, 355–369 (2015). https://doi.org/10.1007/s11103-015-0282-5

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