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

Abstract 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...

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Autores principales:	Xue Zhao, Shishanthi Jayarathna, Helle Turesson, Ann-Sofie Fält, Gustav Nestor, Matías N. González, Niklas Olsson, Mirela Beganovic, Per Hofvander, Roger Andersson, Mariette Andersson
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0a0e1e75316b49b59c1a61b5e0cd52f4
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spelling	oai:doaj.org-article:0a0e1e75316b49b59c1a61b5e0cd52f42021-12-02T11:02:38ZAmylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato10.1038/s41598-021-83462-z2045-2322https://doaj.org/article/0a0e1e75316b49b59c1a61b5e0cd52f42021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83462-zhttps://doaj.org/toc/2045-2322Abstract 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.Xue ZhaoShishanthi JayarathnaHelle TuressonAnn-Sofie FältGustav NestorMatías N. GonzálezNiklas OlssonMirela BeganovicPer HofvanderRoger AnderssonMariette AnderssonNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Xue Zhao Shishanthi Jayarathna Helle Turesson Ann-Sofie Fält Gustav Nestor Matías N. González Niklas Olsson Mirela Beganovic Per Hofvander Roger Andersson Mariette Andersson Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato
description	Abstract 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	article
author	Xue Zhao Shishanthi Jayarathna Helle Turesson Ann-Sofie Fält Gustav Nestor Matías N. González Niklas Olsson Mirela Beganovic Per Hofvander Roger Andersson Mariette Andersson
author_facet	Xue Zhao Shishanthi Jayarathna Helle Turesson Ann-Sofie Fält Gustav Nestor Matías N. González Niklas Olsson Mirela Beganovic Per Hofvander Roger Andersson Mariette Andersson
author_sort	Xue Zhao
title	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_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
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/0a0e1e75316b49b59c1a61b5e0cd52f4
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Amylose starch with no detectable branching developed through DNA-free CRISPR-Cas9 mediated mutagenesis of two starch branching enzymes in potato

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