Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides

PAFs are short cationic and tryptophan-rich synthetic peptides with cell-penetrating antifungal activity. They show potent and selective killing activity against major fungal pathogens and low toxicity to other eukaryotic and bacterial cells. These properties make them a promising alternative to ful...

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Autores: Bundó, Mireia, Shi, Xiaoqing, Vernet, Mar, Marcos López, José Francisco, López García, Belén, Coca, María
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/184036
Acesso em linha:http://hdl.handle.net/10261/184036
Access Level:acceso abierto
Palavra-chave:PAF
Antifungals
Pathogens
Fungi
Rice
Seed
Oil bodies
Protein bodies
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spelling Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptidesBundó, MireiaShi, XiaoqingVernet, MarMarcos López, José FranciscoLópez García, BelénCoca, MaríaPAFAntifungalsPathogensFungiRiceSeedOil bodiesProtein bodiesPAFs are short cationic and tryptophan-rich synthetic peptides with cell-penetrating antifungal activity. They show potent and selective killing activity against major fungal pathogens and low toxicity to other eukaryotic and bacterial cells. These properties make them a promising alternative to fulfill the need of novel antifungals with potential applications in crop protection, food preservation, and medical therapies. However, the difficulties of cost-effective manufacturing of PAFs by chemical synthesis or biotechnological production in microorganisms have hampered their development for practical use. This work explores the feasibility of using rice seeds as an economical and safe production system of PAFs. The rationally designed PAF102 peptide with improved antifungal properties was selected for assessing PAF biotechnological production. Two different strategies are evaluated: (1) the production as a single peptide targeted to protein bodies and (2) the production as an oleosin fusion protein targeted to oil bodies. Both strategies are designed to offer stability to the PAF peptide in the host plant and to facilitate its downstream purification. Our results demonstrate that PAF does not accumulate to detectable levels in rice seeds when produced as a single peptide, whereas it is successfully produced as fusion protein to the Oleosin18, up to 20 μg of peptide per gram of grain. We show that the expression of the chimeric Ole18-PAF102 gene driven by the Ole18 promoter results in the specific accumulation of the fusion protein in the embryo and aleurone layer of the rice seed. Ole18-PAF102 accumulation has no deleterious effects on seed yield, germination capacity, or seedling growth. We also show that the Oleosin18 protein serves as carrier to target the fusion protein to oil bodies facilitating PAF102 recovery. Importantly, the recovered PAF102 is active against the fungal phytopathogen Fusarium proliferatum. Altogether, our results prove that the oleosin fusion technology allows the production of PAF bioactive peptides to assist the exploitation of these antifungal compounds.This work was supported by SEPSAPE grants (Plant KBBE programme) EUI2008-03769 and EUI2008-03619 and by the grant BIO2015-68790-C2-2-R; through the “Severo Ochoa Programme for Centres of Excellence in R&D” (SEV-2015-0533) from Spanish Ministerio de Ciencia, Innovación y Universidades (co-financed FEDER funds); and by the CERCA Programme/Generalitat de Catalunya.Peer reviewedFrontiers MediaGeneralitat de CatalunyaEuropean CommissionMinisterio de Economía y Competitividad (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/184036reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-68790-C2-2-Rinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SEV-2015-0533https://doi.org/10.3389/fpls.2019.00731Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1840362026-05-22T06:33:51Z
dc.title.none.fl_str_mv Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
title Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
spellingShingle Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
Bundó, Mireia
PAF
Antifungals
Pathogens
Fungi
Rice
Seed
Oil bodies
Protein bodies
title_short Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
title_full Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
title_fullStr Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
title_full_unstemmed Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
title_sort Rice seeds as biofactories of rationally designed and cell-penetrating antifungal PAF peptides
dc.creator.none.fl_str_mv Bundó, Mireia
Shi, Xiaoqing
Vernet, Mar
Marcos López, José Francisco
López García, Belén
Coca, María
author Bundó, Mireia
author_facet Bundó, Mireia
Shi, Xiaoqing
Vernet, Mar
Marcos López, José Francisco
López García, Belén
Coca, María
author_role author
author2 Shi, Xiaoqing
Vernet, Mar
Marcos López, José Francisco
López García, Belén
Coca, María
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Generalitat de Catalunya
European Commission
Ministerio de Economía y Competitividad (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv PAF
Antifungals
Pathogens
Fungi
Rice
Seed
Oil bodies
Protein bodies
topic PAF
Antifungals
Pathogens
Fungi
Rice
Seed
Oil bodies
Protein bodies
description PAFs are short cationic and tryptophan-rich synthetic peptides with cell-penetrating antifungal activity. They show potent and selective killing activity against major fungal pathogens and low toxicity to other eukaryotic and bacterial cells. These properties make them a promising alternative to fulfill the need of novel antifungals with potential applications in crop protection, food preservation, and medical therapies. However, the difficulties of cost-effective manufacturing of PAFs by chemical synthesis or biotechnological production in microorganisms have hampered their development for practical use. This work explores the feasibility of using rice seeds as an economical and safe production system of PAFs. The rationally designed PAF102 peptide with improved antifungal properties was selected for assessing PAF biotechnological production. Two different strategies are evaluated: (1) the production as a single peptide targeted to protein bodies and (2) the production as an oleosin fusion protein targeted to oil bodies. Both strategies are designed to offer stability to the PAF peptide in the host plant and to facilitate its downstream purification. Our results demonstrate that PAF does not accumulate to detectable levels in rice seeds when produced as a single peptide, whereas it is successfully produced as fusion protein to the Oleosin18, up to 20 μg of peptide per gram of grain. We show that the expression of the chimeric Ole18-PAF102 gene driven by the Ole18 promoter results in the specific accumulation of the fusion protein in the embryo and aleurone layer of the rice seed. Ole18-PAF102 accumulation has no deleterious effects on seed yield, germination capacity, or seedling growth. We also show that the Oleosin18 protein serves as carrier to target the fusion protein to oil bodies facilitating PAF102 recovery. Importantly, the recovered PAF102 is active against the fungal phytopathogen Fusarium proliferatum. Altogether, our results prove that the oleosin fusion technology allows the production of PAF bioactive peptides to assist the exploitation of these antifungal compounds.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/184036
url http://hdl.handle.net/10261/184036
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-68790-C2-2-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SEV-2015-0533
https://doi.org/10.3389/fpls.2019.00731
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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