TY - JOUR

T1 - Reduction in Cold-Induced Sweetening by Cas9 Endonuclease-Mediated Knockout of the POTATO VACUOLAR INVERTASE 1 Gene in the Cultivar ‘Symfonia’

AU - Egorova, Anastasiya A.

AU - Zykova, Tatyana E.

AU - Kostina, Nina E.

AU - Saboiev, Izatsho A.

AU - Koloshina, Kristina A.

AU - Filipenko, Elena A.

AU - Hoffie, Iris

AU - Hertig, Christian

AU - Hiekel, Stefan

AU - Kumlehn, Jochen

AU - Kochetov, Alex V.

AU - Gerasimova, Sophia V.

N1 - The study is supported by the ICG Kurchatov Genomics Centre (075–15-2019–1662).

PY - 2024/10/5

Y1 - 2024/10/5

N2 - Storage of potato tubers at low temperatures leads to the degradation of starch into reducing sugars, resulting in cold-induced sweetening (CIS). These sugars react with free amino acids when processed at high temperatures, resulting in brown, bitter-tasting products. Sucrose is one of the intermediate steps of conversion of starch into reducing sugars. Vacuolar invertase breaks down sucrose to glucose and fructose, thereby largely contributing to the accumulation of reducing sugars. Targeted knockout of the potato vacuolar invertase (Pain-1) gene by Cas9/gRNA-mediated site-directed mutagenesis was performed in the tetraploid potato cultivar Symfonia. A vector with expression units for cas9 and three Pain-1-specific gRNAs was constructed, and its mutagenic activity demonstrated in potato protoplasts. Eleven mutant lines were then obtained by Agrobacterium-mediated transformation, with nine of them carrying mutations in all four gene copies present in tetraploid potato. Tubers from four Pain-1 mutant lines were analyzed for sugar accumulation during cold storage. The two lines with knockout of all four gene copies had significantly reduced glucose and fructose levels and lighter chip color compared to the non-mutated control line. This work demonstrates that CIS resistance can be established in a potato elite cultivar by targeted Pain-1 mutagenesis.

AB - Storage of potato tubers at low temperatures leads to the degradation of starch into reducing sugars, resulting in cold-induced sweetening (CIS). These sugars react with free amino acids when processed at high temperatures, resulting in brown, bitter-tasting products. Sucrose is one of the intermediate steps of conversion of starch into reducing sugars. Vacuolar invertase breaks down sucrose to glucose and fructose, thereby largely contributing to the accumulation of reducing sugars. Targeted knockout of the potato vacuolar invertase (Pain-1) gene by Cas9/gRNA-mediated site-directed mutagenesis was performed in the tetraploid potato cultivar Symfonia. A vector with expression units for cas9 and three Pain-1-specific gRNAs was constructed, and its mutagenic activity demonstrated in potato protoplasts. Eleven mutant lines were then obtained by Agrobacterium-mediated transformation, with nine of them carrying mutations in all four gene copies present in tetraploid potato. Tubers from four Pain-1 mutant lines were analyzed for sugar accumulation during cold storage. The two lines with knockout of all four gene copies had significantly reduced glucose and fructose levels and lighter chip color compared to the non-mutated control line. This work demonstrates that CIS resistance can be established in a potato elite cultivar by targeted Pain-1 mutagenesis.

KW - CRISPR/Cas

KW - Gene editing

KW - Potato chips

KW - Sugars

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85205039054&origin=inward&txGid=ba77acea6fc07bf97a1ac76547bdbbda

UR - https://www.webofscience.com/wos/woscc/full-record/WOS:001324527700001

UR - https://www.mendeley.com/catalogue/b0c1b1a4-ae0d-3a86-b34d-fd4eb899df59/

U2 - 10.1007/s11540-024-09800-6

DO - 10.1007/s11540-024-09800-6

M3 - Article

JO - Potato Research

JF - Potato Research

SN - 0014-3065

ER -