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High expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants

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Abstract

Chloroplast transformation has an extraordinary potential for antigen production in plants because of the capacity to accumulate high levels of recombinant proteins and increased biosafety due to maternal plastid inheritance in most crops. In this article, we evaluate tobacco chloroplasts transformation for the production of a highly immunogenic epitope containing amino acid residues 135–160 of the structural protein VP1 of the foot and mouth disease virus (FMDV). To increase the accumulation levels, the peptide was expressed as a fusion protein with the β-glucuronidase reporter gene (uidA). The recombinant protein represented the 51% of the total soluble proteins in mature leaves, a level higher than those of the Rubisco large subunit, the most abundant protein in the leaf of a wild-type plant. Despite this high accumulation of heterologous protein, the transplastomic plants and wild-type tobacco were phenotypically indistinguishable. The FMDV epitope expressed in transplastomic plants was immunogenic in mice. These results show that transplastomic tobacco express efficiently the recombinant protein, and we conclude that this technology allows the production of large quantities of immunogenic proteins.

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Abbreviations

FMDV:

Foot and mouth disease virus

ELISA:

Enzyme-linked immunosorbent assay

TSP:

Total soluble proteins

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Acknowledgments

We wish to thank Demian Bellido and Fatima Torales for their technical assistance and Carolina Pometti for carefully reading the manuscript. This work has been supported by Instituto Nacional de Tecnología Agropecuaria (INTA-Argentina), project “Fiebre Aftosa”, no. AESA 1572. EML and MES are fellows of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-Argentina). MMM is assistant teacher of Universidad de Buenos Aires, Argentina. FBA and SAW are research scientists of CONICET, MJDS and AW are research scientists of CONICET and INTA, and MVM is research scientist of INTA.

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Authors and Affiliations

  1. Laboratorio de Virología y Biotecnología Vegetal, INGEBI-UBA/CONICET, Ciudad Autónoma de Buenos Aires, Vuelta de Obligado 2490, C1428ADN, Buenos Aires, Argentina

    Ezequiel Matías Lentz, María Eugenia Segretin, Mauro Miguel Morgenfeld, Sonia Alejandra Wirth & Fernando Félix Bravo-Almonacid

  2. Instituto de Virología, CICV y A, INTA, Los Reseros y las Cabañas S/N, Castelar, Buenos Aires, Argentina

    María José Dus Santos, Marina Valeria Mozgovoj & Andrés Wigdorovitz

Authors
  1. Ezequiel Matías Lentz
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  2. María Eugenia Segretin
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  3. Mauro Miguel Morgenfeld
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  4. Sonia Alejandra Wirth
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  5. María José Dus Santos
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  6. Marina Valeria Mozgovoj
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  7. Andrés Wigdorovitz
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  8. Fernando Félix Bravo-Almonacid
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Correspondence to Fernando Félix Bravo-Almonacid.

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Lentz, E.M., Segretin, M.E., Morgenfeld, M.M. et al. High expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants. Planta 231, 387–395 (2010). https://doi.org/10.1007/s00425-009-1058-4

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