Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
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%...
| Main Authors: | , , , , , , , , , , |
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
| Published: |
Nature Publishing Group
2021
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| Subjects: | |
| Online Access: | 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 |
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| author | 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_browse | Anderson, Mariette Andersson, Roger Beganovic, Mirela Fält, Ann Sofie Gonzalez, Matías Nicolás Hofvander, Per Jayarathna, Shishanthi Nestor, Gustav Olsson, Niklas Turesson, Helle 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_sort | Zhao, Xue |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA9235 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Nature Publishing Group |
| publisherStr | Nature Publishing Group |
| record_format | dspace |
| spelling | INTA92352021-04-30T11:26:55Z Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato Zhao, Xue Jayarathna, Shishanthi Turesson, Helle Fält, Ann Sofie Nestor, Gustav Gonzalez, Matías Nicolás Olsson, Niklas Beganovic, Mirela Hofvander, Per Andersson, Roger Anderson, Mariette Papa Enzimas Almidón Amilosa Amilopectinia Potatoes Enzymes Genes Mutagenesis Starch Amilose Amylopectin 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. 2021-04-30T11:17:52Z 2021-04-30T11:17:52Z 2021-02-22 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf Nature Publishing Group Scientific Reports 11 : 4311 (2021) |
| spellingShingle | Papa Enzimas Almidón Amilosa Amilopectinia Potatoes Enzymes Genes Mutagenesis Starch Amilose Amylopectin Zhao, Xue Jayarathna, Shishanthi Turesson, Helle Fält, Ann Sofie Nestor, Gustav Gonzalez, Matías Nicolás Olsson, Niklas Beganovic, Mirela Hofvander, Per Andersson, Roger Anderson, Mariette 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 |
| 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_short | 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 |
| topic | Papa Enzimas Almidón Amilosa Amilopectinia Potatoes Enzymes Genes Mutagenesis Starch Amilose Amylopectin |
| 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 |
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