Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato

Autores
Zhao, Xue; Jayarathna, Shishanthi; Turesson, Helle; Fält, Ann Sofie; Nestor, Gustav; Gonzalez, Matías Nicolás; Olsson, Niklas; Beganovic, Mirela; Hofvander, Per; Andersson, Roger; Andersson, Mariette
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
DNA-free genome editing was used to induce mutations in one or two branching enzyme genes (Sbe) in tetraploid potato to develop starch with an increased amylose ratio and elongated amylopectin chains. By using ribonucleoprotein (RNP) transfection of potato protoplasts, a mutation frequency up to 72% was achieved. The large variation of mutations was grouped as follows: Group 1 lines with all alleles of Sbe1 mutated, Group 2 lines with all alleles of Sbe1 as well as two to three alleles of Sbe2 mutated and Group 3 lines having all alleles of both genes mutated. Starch from lines in Group 3 was found to be essentially free of amylopectin with no detectable branching and a chain length (CL) distribution where not only the major amylopectin fraction but also the shortest amylose chains were lost. Surprisingly, the starch still formed granules in a low-ordered crystalline structure. Starch from lines of Group 2 had an increased CL with a higher proportion of intermediate-sized chains, an altered granule phenotype but a crystalline structure in the granules similar to wild-type starch. Minor changes in CL could also be detected for the Group 1 starches when studied at a higher resolution.
Fil: Zhao, Xue. Swedish University of Agricultural Sciences; Suecia
Fil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences; Suecia
Fil: Turesson, Helle. Swedish University of Agricultural Sciences; Suecia
Fil: Fält, Ann Sofie. Swedish University of Agricultural Sciences; Suecia
Fil: Nestor, Gustav. Swedish University of Agricultural Sciences; Suecia
Fil: Gonzalez, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences; Suecia
Fil: Beganovic, Mirela. Swedish University of Agricultural Sciences; Suecia
Fil: Hofvander, Per. Swedish University of Agricultural Sciences; Suecia
Fil: Andersson, Roger. Swedish University of Agricultural Sciences; Suecia
Fil: Andersson, Mariette. Swedish University of Agricultural Sciences; Suecia
Materia
CRISPR/CAS9
STARCH BRANCHING ENZYMES
AMYLOSE
POTATO
RIBONUCLEOPROTEIN COMPLEXES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/176695
Acceder
id CONICETDig_ff593aaca204a0f786a4776660f40d11
oai_identifier_str oai:ri.conicet.gov.ar:11336/176695
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potatoZhao, XueJayarathna, ShishanthiTuresson, HelleFält, Ann SofieNestor, GustavGonzalez, Matías NicolásOlsson, NiklasBeganovic, MirelaHofvander, PerAndersson, RogerAndersson, MarietteCRISPR/CAS9STARCH BRANCHING ENZYMESAMYLOSEPOTATORIBONUCLEOPROTEIN COMPLEXEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1DNA-free genome editing was used to induce mutations in one or two branching enzyme genes (Sbe) in tetraploid potato to develop starch with an increased amylose ratio and elongated amylopectin chains. By using ribonucleoprotein (RNP) transfection of potato protoplasts, a mutation frequency up to 72% was achieved. The large variation of mutations was grouped as follows: Group 1 lines with all alleles of Sbe1 mutated, Group 2 lines with all alleles of Sbe1 as well as two to three alleles of Sbe2 mutated and Group 3 lines having all alleles of both genes mutated. Starch from lines in Group 3 was found to be essentially free of amylopectin with no detectable branching and a chain length (CL) distribution where not only the major amylopectin fraction but also the shortest amylose chains were lost. Surprisingly, the starch still formed granules in a low-ordered crystalline structure. Starch from lines of Group 2 had an increased CL with a higher proportion of intermediate-sized chains, an altered granule phenotype but a crystalline structure in the granules similar to wild-type starch. Minor changes in CL could also be detected for the Group 1 starches when studied at a higher resolution.Fil: Zhao, Xue. Swedish University of Agricultural Sciences; SueciaFil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences; SueciaFil: Turesson, Helle. Swedish University of Agricultural Sciences; SueciaFil: Fält, Ann Sofie. Swedish University of Agricultural Sciences; SueciaFil: Nestor, Gustav. Swedish University of Agricultural Sciences; SueciaFil: Gonzalez, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Olsson, Niklas. Swedish University of Agricultural Sciences; SueciaFil: Beganovic, Mirela. Swedish University of Agricultural Sciences; SueciaFil: Hofvander, Per. Swedish University of Agricultural Sciences; SueciaFil: Andersson, Roger. Swedish University of Agricultural Sciences; SueciaFil: Andersson, Mariette. Swedish University of Agricultural Sciences; SueciaNature Publishing Group2021-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/176695Zhao, Xue; Jayarathna, Shishanthi; Turesson, Helle; Fält, Ann Sofie; Nestor, Gustav; et al.; Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato; Nature Publishing Group; Scientific Reports; 11; 1; 2-2021; 1-132045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-021-83462-zinfo:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-021-83462-zinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:39:35Zoai:ri.conicet.gov.ar:11336/176695instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:39:35.687CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
title Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
spellingShingle Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
Zhao, Xue
CRISPR/CAS9
STARCH BRANCHING ENZYMES
AMYLOSE
POTATO
RIBONUCLEOPROTEIN COMPLEXES
title_short Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
title_full Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
title_fullStr Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
title_full_unstemmed Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
title_sort Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
dc.creator.none.fl_str_mv Zhao, Xue
Jayarathna, Shishanthi
Turesson, Helle
Fält, Ann Sofie
Nestor, Gustav
Gonzalez, Matías Nicolás
Olsson, Niklas
Beganovic, Mirela
Hofvander, Per
Andersson, Roger
Andersson, Mariette
author Zhao, Xue
author_facet Zhao, Xue
Jayarathna, Shishanthi
Turesson, Helle
Fält, Ann Sofie
Nestor, Gustav
Gonzalez, Matías Nicolás
Olsson, Niklas
Beganovic, Mirela
Hofvander, Per
Andersson, Roger
Andersson, Mariette
author_role author
author2 Jayarathna, Shishanthi
Turesson, Helle
Fält, Ann Sofie
Nestor, Gustav
Gonzalez, Matías Nicolás
Olsson, Niklas
Beganovic, Mirela
Hofvander, Per
Andersson, Roger
Andersson, Mariette
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CRISPR/CAS9
STARCH BRANCHING ENZYMES
AMYLOSE
POTATO
RIBONUCLEOPROTEIN COMPLEXES
topic CRISPR/CAS9
STARCH BRANCHING ENZYMES
AMYLOSE
POTATO
RIBONUCLEOPROTEIN COMPLEXES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv DNA-free genome editing was used to induce mutations in one or two branching enzyme genes (Sbe) in tetraploid potato to develop starch with an increased amylose ratio and elongated amylopectin chains. By using ribonucleoprotein (RNP) transfection of potato protoplasts, a mutation frequency up to 72% was achieved. The large variation of mutations was grouped as follows: Group 1 lines with all alleles of Sbe1 mutated, Group 2 lines with all alleles of Sbe1 as well as two to three alleles of Sbe2 mutated and Group 3 lines having all alleles of both genes mutated. Starch from lines in Group 3 was found to be essentially free of amylopectin with no detectable branching and a chain length (CL) distribution where not only the major amylopectin fraction but also the shortest amylose chains were lost. Surprisingly, the starch still formed granules in a low-ordered crystalline structure. Starch from lines of Group 2 had an increased CL with a higher proportion of intermediate-sized chains, an altered granule phenotype but a crystalline structure in the granules similar to wild-type starch. Minor changes in CL could also be detected for the Group 1 starches when studied at a higher resolution.
Fil: Zhao, Xue. Swedish University of Agricultural Sciences; Suecia
Fil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences; Suecia
Fil: Turesson, Helle. Swedish University of Agricultural Sciences; Suecia
Fil: Fält, Ann Sofie. Swedish University of Agricultural Sciences; Suecia
Fil: Nestor, Gustav. Swedish University of Agricultural Sciences; Suecia
Fil: Gonzalez, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences; Suecia
Fil: Beganovic, Mirela. Swedish University of Agricultural Sciences; Suecia
Fil: Hofvander, Per. Swedish University of Agricultural Sciences; Suecia
Fil: Andersson, Roger. Swedish University of Agricultural Sciences; Suecia
Fil: Andersson, Mariette. Swedish University of Agricultural Sciences; Suecia
description DNA-free genome editing was used to induce mutations in one or two branching enzyme genes (Sbe) in tetraploid potato to develop starch with an increased amylose ratio and elongated amylopectin chains. By using ribonucleoprotein (RNP) transfection of potato protoplasts, a mutation frequency up to 72% was achieved. The large variation of mutations was grouped as follows: Group 1 lines with all alleles of Sbe1 mutated, Group 2 lines with all alleles of Sbe1 as well as two to three alleles of Sbe2 mutated and Group 3 lines having all alleles of both genes mutated. Starch from lines in Group 3 was found to be essentially free of amylopectin with no detectable branching and a chain length (CL) distribution where not only the major amylopectin fraction but also the shortest amylose chains were lost. Surprisingly, the starch still formed granules in a low-ordered crystalline structure. Starch from lines of Group 2 had an increased CL with a higher proportion of intermediate-sized chains, an altered granule phenotype but a crystalline structure in the granules similar to wild-type starch. Minor changes in CL could also be detected for the Group 1 starches when studied at a higher resolution.
publishDate 2021
dc.date.none.fl_str_mv 2021-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/176695
Zhao, Xue; Jayarathna, Shishanthi; Turesson, Helle; Fält, Ann Sofie; Nestor, Gustav; et al.; Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato; Nature Publishing Group; Scientific Reports; 11; 1; 2-2021; 1-13
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/176695
identifier_str_mv Zhao, Xue; Jayarathna, Shishanthi; Turesson, Helle; Fält, Ann Sofie; Nestor, Gustav; et al.; Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato; Nature Publishing Group; Scientific Reports; 11; 1; 2-2021; 1-13
2045-2322
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-021-83462-z
info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-021-83462-z
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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