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; Anderson, 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.
EEA Balcarce
Fil: Zhao, Xue. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Turesson, Helle. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Fält , Ann Sofie. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Nestor, Gustav. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: González, Matías Nicolás. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina.
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Beganovic, Mirela. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Hofvander, Per. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Andersson, Roger. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Anderson, Mariette. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fuente
Scientific Reports 11 : 4311 (2021)
Materia
Papa
Enzimas
Almidón
Amilosa
Amilopectinia
Potatoes
Enzymes
Genes
Mutagenesis
Starch
Amilose
Amylopectin
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/9235
Acceder
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oai_identifier_str oai:localhost:20.500.12123/9235
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
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, RogerAnderson, MariettePapaEnzimasAlmidónAmilosaAmilopectiniaPotatoesEnzymesGenesMutagenesisStarchAmiloseAmylopectinDNA-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.EEA BalcarceFil: Zhao, Xue. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.Fil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.Fil: Turesson, Helle. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Fil: Fält , Ann Sofie. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Fil: Nestor, Gustav. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.Fil: González, Matías Nicolás. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina.Fil: Olsson, Niklas. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Fil: Beganovic, Mirela. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Fil: Hofvander, Per. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Fil: Andersson, Roger. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.Fil: Anderson, Mariette. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.Nature Publishing Group2021-04-30T11:17:52Z2021-04-30T11:17:52Z2021-02-22info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/9235https://www.nature.com/articles/s41598-021-83462-z2045-2322 (online)https://doi.org/10.1038/s41598-021-83462-zScientific Reports 11 : 4311 (2021)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:45:12Zoai:localhost:20.500.12123/9235instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:45:12.622INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
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
Papa
Enzimas
Almidón
Amilosa
Amilopectinia
Potatoes
Enzymes
Genes
Mutagenesis
Starch
Amilose
Amylopectin
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
Anderson, 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
Anderson, 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
Anderson, Mariette
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Papa
Enzimas
Almidón
Amilosa
Amilopectinia
Potatoes
Enzymes
Genes
Mutagenesis
Starch
Amilose
Amylopectin
topic Papa
Enzimas
Almidón
Amilosa
Amilopectinia
Potatoes
Enzymes
Genes
Mutagenesis
Starch
Amilose
Amylopectin
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.
EEA Balcarce
Fil: Zhao, Xue. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Jayarathna, Shishanthi. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Turesson, Helle. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Fält , Ann Sofie. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Nestor, Gustav. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: González, Matías Nicolás. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina.
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Beganovic, Mirela. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Hofvander, Per. Swedish University of Agricultural Sciences. Department of Plant Breeding; Suecia.
Fil: Andersson, Roger. Swedish University of Agricultural Sciences. Department of Molecular Sciences; Suecia.
Fil: Anderson, Mariette. Swedish University of Agricultural Sciences. Department of Plant Breeding; 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 2021-04-30T11:17:52Z
2021-04-30T11:17:52Z
2021-02-22
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/20.500.12123/9235
https://www.nature.com/articles/s41598-021-83462-z
2045-2322 (online)
https://doi.org/10.1038/s41598-021-83462-z
url http://hdl.handle.net/20.500.12123/9235
https://www.nature.com/articles/s41598-021-83462-z
https://doi.org/10.1038/s41598-021-83462-z
identifier_str_mv 2045-2322 (online)
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv 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 Scientific Reports 11 : 4311 (2021)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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score 12.559606

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