Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut

Castanea sativa, a species of high ecological and economic relevance in Europe, faces severe threats from root rot caused by Phytophthora cinnamomi. To explore genetic strategies for enhancing disease tolerance, we investigated the functional role of a chestnut gene homologous to Ginkgo biloba'...

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Autores: Serrazina, Susana, Martínez-Santiago, María Teresa, Valladares, Silvia, Del Castillo-González, Lucía, Francisco, Marcelo, Berrocal-Lobo, Marta, Piñas, Eduardo, Piñeiro, Pablo, Malhó, Rui, Costa, Rita Lourenço, Corredoira, Elena
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2026
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/417664
Acceso en línea:http://hdl.handle.net/10261/417664
Access Level:acceso abierto
Palabra clave:Agrobacterium tumefaciens
Castanea sativa
Cysteine-rich repeat secretory protein
Genetic transformation
In vitro tolerance assays
Ink disease
Somatic embryos
http://metadata.un.org/sdg/15
Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat desertification, and halt and reverse land degradation and halt biodiversity loss
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spelling Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnutSerrazina, SusanaMartínez-Santiago, María TeresaValladares, SilviaDel Castillo-González, LucíaFrancisco, MarceloBerrocal-Lobo, MartaPiñas, EduardoPiñeiro, PabloMalhó, RuiCosta, Rita LourençoCorredoira, ElenaAgrobacterium tumefaciensCastanea sativaCysteine-rich repeat secretory proteinGenetic transformationIn vitro tolerance assaysInk diseaseSomatic embryoshttp://metadata.un.org/sdg/15Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat desertification, and halt and reverse land degradation and halt biodiversity lossCastanea sativa, a species of high ecological and economic relevance in Europe, faces severe threats from root rot caused by Phytophthora cinnamomi. To explore genetic strategies for enhancing disease tolerance, we investigated the functional role of a chestnut gene homologous to Ginkgo biloba's ginkbilobin-2 (Cast_Gnk2-like), known for its antifungal properties. Using Agrobacterium tumefaciens-mediated transformation, the Cast_Gnk2-like gene was introduced into somatic embryos from two embryogenic chestnut lines. Transformation efficiency was genotype-dependent, and varied from 14.2% to 2.5%. Twelve independent transgenic lines were confirmed by PCR, and each was estimated to carry a single copy of the transgene. Gene expression analysis revealed significant Cast_Gnk2-like transcript levels in two transgenic lines. Following cold storage and germination treatment, viable transgenic plants were regenerated. Disease tolerance assays demonstrated that Cast_Gnk2-like overexpression significantly reduced root necrosis and symptom severity, indicating enhanced tolerance to P. cinnamomi. These findings highlight the potential of targeted gene overexpression to improve disease resilience in chestnut through genetic engineering.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. At MBG-CSIC, work was partly supported by Agencia Estatal de Investigación (AEI, Spain) through the projects PID2020-112627RB-C33 (AEI/https://doi.org/10.13039/501100011033) and PID2024-156422OB-C33. At BioISI, work was supported by UIDB/04046/2020 (DOI: https://doi.org/10.54499/UIDB/ 04046/ 2020) and UIDP/04046/2020 (DOI: https://doi.org/10.54499/UIDP/ 04046/ 2020) Centre grants from Fundação para a Ciência e a Tecnologia, Portugal. At UPM, research was partially funded by EU FEADER funds through the project “Breeding and conservation program for holm and cork oak for tolerance to oak decline” leaded by the Spanish Ministry for Ecological Transition and Demographic Challenge (MITECO, Ministerio de Agricultura, Pesca y Alimentación). At INIAV this work was supported by FCT – Fundação para a Ciência e Tecnologia, I.P., through project reference UID/00239/2025. Portugal. SS is being funded by an FCT contract in the project 2022.06990. PTDC (DOI: https://doi.org/10.54499/2022.06990.PTDC). LDCG was supported by Fundación Conde Del Valle Salazar (UPM, Spain) and by Fucovasa Contract forming part of her PhD directed by Berrocal-Lobo (UPM). PP was funded by a FPU (Formación de Profesorado Universitario) grant from Ministerio de Ciencia, Innovación y Universidades (Spain) (FPU23/00921). We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).Peer reviewedSpringer NatureConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202620262026info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/417664reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésBMC genomicshttps://doi.org/10.1186/s12864-025-12485-xSíinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4176642026-05-22T06:33:51Z
dc.title.none.fl_str_mv Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
title Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
spellingShingle Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
Serrazina, Susana
Agrobacterium tumefaciens
Castanea sativa
Cysteine-rich repeat secretory protein
Genetic transformation
In vitro tolerance assays
Ink disease
Somatic embryos
http://metadata.un.org/sdg/15
Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat desertification, and halt and reverse land degradation and halt biodiversity loss
title_short Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
title_full Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
title_fullStr Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
title_full_unstemmed Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
title_sort Overexpression of ginkbilobin-2 homologous domain gene to enhance the tolerance to Phytophthora cinnamomi in plants of European chestnut
dc.creator.none.fl_str_mv Serrazina, Susana
Martínez-Santiago, María Teresa
Valladares, Silvia
Del Castillo-González, Lucía
Francisco, Marcelo
Berrocal-Lobo, Marta
Piñas, Eduardo
Piñeiro, Pablo
Malhó, Rui
Costa, Rita Lourenço
Corredoira, Elena
author Serrazina, Susana
author_facet Serrazina, Susana
Martínez-Santiago, María Teresa
Valladares, Silvia
Del Castillo-González, Lucía
Francisco, Marcelo
Berrocal-Lobo, Marta
Piñas, Eduardo
Piñeiro, Pablo
Malhó, Rui
Costa, Rita Lourenço
Corredoira, Elena
author_role author
author2 Martínez-Santiago, María Teresa
Valladares, Silvia
Del Castillo-González, Lucía
Francisco, Marcelo
Berrocal-Lobo, Marta
Piñas, Eduardo
Piñeiro, Pablo
Malhó, Rui
Costa, Rita Lourenço
Corredoira, Elena
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Agrobacterium tumefaciens
Castanea sativa
Cysteine-rich repeat secretory protein
Genetic transformation
In vitro tolerance assays
Ink disease
Somatic embryos
http://metadata.un.org/sdg/15
Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat desertification, and halt and reverse land degradation and halt biodiversity loss
topic Agrobacterium tumefaciens
Castanea sativa
Cysteine-rich repeat secretory protein
Genetic transformation
In vitro tolerance assays
Ink disease
Somatic embryos
http://metadata.un.org/sdg/15
Protect, restore and promote sustainable use of terrestrial ecosystems, sustainably manage forests, combat desertification, and halt and reverse land degradation and halt biodiversity loss
description Castanea sativa, a species of high ecological and economic relevance in Europe, faces severe threats from root rot caused by Phytophthora cinnamomi. To explore genetic strategies for enhancing disease tolerance, we investigated the functional role of a chestnut gene homologous to Ginkgo biloba's ginkbilobin-2 (Cast_Gnk2-like), known for its antifungal properties. Using Agrobacterium tumefaciens-mediated transformation, the Cast_Gnk2-like gene was introduced into somatic embryos from two embryogenic chestnut lines. Transformation efficiency was genotype-dependent, and varied from 14.2% to 2.5%. Twelve independent transgenic lines were confirmed by PCR, and each was estimated to carry a single copy of the transgene. Gene expression analysis revealed significant Cast_Gnk2-like transcript levels in two transgenic lines. Following cold storage and germination treatment, viable transgenic plants were regenerated. Disease tolerance assays demonstrated that Cast_Gnk2-like overexpression significantly reduced root necrosis and symptom severity, indicating enhanced tolerance to P. cinnamomi. These findings highlight the potential of targeted gene overexpression to improve disease resilience in chestnut through genetic engineering.
publishDate 2026
dc.date.none.fl_str_mv 2026
2026
2026
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/10261/417664
url http://hdl.handle.net/10261/417664
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv BMC genomics
https://doi.org/10.1186/s12864-025-12485-x
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
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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