Efficient production of antifungal proteins in plants using a new transient expression vector derived from tobacco mosaic virus
Shi, Xiaoqing (Centre de Recerca en Agrigenòmica)
Cordero, Teresa (Instituto de Biología Molecular y Celular de Plantas)
Garrigues, Sandra (Consejo Superior de Investigaciones Científicas (Espanya). Instituto de Agroquimica y Tecnologia de Alimentos)
Marcos, Jose F. (Consejo Superior de Investigaciones Científicas (Espanya). Instituto de Agroquimica y Tecnologia de Alimentos)
Daròs, José-Antonio (Instituto de Biología Molecular y Celular de Plantas)
Coca López, María (Centre de Recerca en Agrigenòmica)

Fecha:	2019
Resumen:	Fungi that infect plants, animals or humans pose a serious threat to human health and food security. Antifungal proteins (AFPs) secreted by filamentous fungi are promising biomolecules that could be used to develop new antifungal therapies in medicine and agriculture. They are small highly stable proteins with specific potent activity against fungal pathogens. However, their exploitation requires efficient, sustainable and safe production systems. Here, we report the development of an easy-to-use, open access viral vector based on Tobacco mosaic virus (TMV). This new system allows the fast and efficient assembly of the open reading frames of interest in small intermediate entry plasmids using the Gibson reaction. The manipulated TMV fragments are then transferred to the infectious clone by a second Gibson assembly reaction. Recombinant proteins are produced by agroinoculating plant leaves with the resulting infectious clones. Using this simple viral vector, we have efficiently produced two different AFPs in Nicotiana benthamiana leaves, namely the Aspergillus giganteus AFP and the Penicillium digitatum AfpB. We obtained high protein yields by targeting these bioactive small proteins to the apoplastic space of plant cells. However, when AFPs were targeted to intracellular compartments, we observed toxic effects in the host plants and undetectable levels of protein. We also demonstrate that this production system renders AFPs fully active against target pathogens, and that crude plant extracellular fluids containing the AfpB can protect tomato plants from Botrytis cinerea infection, thus supporting the idea that plants are suitable biofactories to bring these antifungal proteins to the market.
Ayudas:	Ministerio de Ciencia e Innovación BIO2017-83184-R Ministerio de Ciencia e Innovación BIO2015-68790-C2-1-R Ministerio de Ciencia e Innovación BIO2015-68790-C2-2-R Ministerio de Ciencia e Innovación SEV-2015-0533
Nota:	Altres ajuts: CERCA Programme/Generalitat de Catalunya
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Antifungal proteins ; Gibson assembly ; Nicotiana benthamiana ; Plant biofactory ; Tobacco mosaic virus ; Viral vector
Publicado en:	Plant Biotechnology Journal, Vol. 17, Issue 6 (June 2019) , p. 1069-1080, ISSN 1467-7652

DOI: 10.1111/pbi.13038
PMID: 30521145

12 p, 6.1 MB

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Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias > CRAG (Centro de Investigación en Agrigenómica)
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