Potato genome editing using the CRISPR/Cas9 system to knock out the StDMR6-1 and StCHL1 genes involved in late blight resistance
https://doi.org/10.29235/1029-8940-2026-71-2-104-115
Abstract
Using Agrobacterium-mediated transformation, vector genetic constructs, carrying CRISPR/Cas9 elements in their composition for the knockout of the StDMR6-1 and StCHL1 genes regulating defense responses, were introduced into the genomes of Belarusian potato varieties Pershatsvet, Yuliya and Krasavik. The knockout of these genes (S-genes) is a promising, essential and modern approach for increasing potato resistance to late blight. A total of 288 transgenic T0 potato plants were obtained, and 161 out of them were analyzed for insertion-deletion mutations using Sanger sequencing. Moreover, out of 161 transformants of the T0 generation, 84 had mutant sequences of the StCHL1 and StDMR6-1 genes with a mutation frequency from 1 to 97 % at p < 0.001 and 99.2 % at p ≥ 0.001, depending on the variety. As a result of the experiment, genetically edited potato plants of the varieties of Belarusian selection Yuliya, Pershatsvet, and Krasavik were obtained for the first time in the Republic of Belarus. These plants carry mutations in the StCHL1 and StDMR6-1 genes, leading to a shift in the reading frame and, as a consequence, to gene knockout.
Supporting agencies: The work was carried out within the framework of a separate project of fundamental and applied research of the National Academy of Sciences of Belarus “To create potato plants carrying genetically edited using the CRISPR/Cas9 system genes regulating defense reactions, StDMR6-1 and StCHL1, to increase resistance to late blight” and with the support of the BRFFR grant No. B23-062.
About the Authors
A. M. Shishlova
Institute of Genetics and Cytology of the National Academy of Sciences of Belarus
Belarus
Belarus
Anastasia M. Shishlova-Sokolovskaya – Senior Researcher
34, F. Skorina Str., 220141, Minsk
V. E. Neborskaya
Institute of Genetics and Cytology of the National Academy of Sciences of Belarus
Belarus
Belarus
Valeria E. Neborskaya – Junior Researcher
34, F. Skorina Str., 220141, Minsk
O. Yu. Urbanovich
Institute of Genetics and Cytology of the National Academy of Sciences of Belarus
Belarus
Belarus
Oksana Yu. Urbanovich – D. Sc. (Biol.), Professor, Head of the Laboratory
34, F. Skorina Str., 220141, Minsk
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