Accumulation of Anthocyanin in the Aleurone of Barley Grains by Targeted Restoration of the MYC2 Gene

Accumulation of Anthocyanin in the Aleurone of Barley Grains by Targeted Restoration of the MYC2 Gene Full article

Journal

International Journal of Molecular Sciences
ISSN: 1661-6596

Output data

Year: 2024, Volume: 25, Number: 23, Article number : 12705, Pages count : DOI: 10.3390/ijms252312705

Tags

genome editing; Cas9/gRNA; pigmented barley; nutritional value; flavonoid biosynthesis; grain color

Authors

Egorova Anastasiya A. ^1,2,3,4 , Zykova Tatyana E. ^1,2,4 , Hertig Christian W. ³ , Hoffie Iris ³ , Morozov Sergey V. ^2,5 , Chernyak Elena I. ⁵ , Rogachev Artem D. ⁵ , Korotkova Anna M. ^1,4 , Vikhorev Alexander V. ² , Vasiliev Gennady V. ¹ , Shoeva Olesya Y. ^1,2,4 , Kumlehn Jochen ³ , Gerasimova Sophia V. ^1,3,4 , Khlestkina Elena K. ^1,4

Affiliations

1	Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences
2	Faculty of Natural Sciences, Novosibirsk State University
3	Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)
4	N.I. Vavilov All-Russian Research Institute of Plant Genetic Resources
5	N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences

Blue barley grain pigmentation results from anthocyanin accumulation in the aleurone layer. Anthocyanins are known for their beneficial effects on human health. The gene encoding the MYELOCYTOMATOSIS 2 (MYC2) transcription factor is potentially responsible for the blue coloration of the aleurone. In non-pigmented barley, a single nucleotide insertion in this gene causes a frameshift mutation with a premature stop codon. It was hypothesized that restoring the MYC2 reading frame could activate anthocyanin accumulation in the aleurone. Using a targeted mutagenesis approach in the present study, the reading frame of MYC2 was restored in the non-pigmented cultivar Golden Promise. Genetic constructs harboring cas9 and gRNA expression units were developed, pre-validated in protoplasts, and then functional MYC2 alleles were generated at the plant level via Agrobacterium-mediated transformation. Anthocyanin accumulation in the aleurone layer of grains from these mutants was confirmed through microscopy and chemical analysis. The expression of anthocyanin biosynthesis genes was analyzed, revealing that the restoration of MYC2 led to increased transcript levels of F3H and ANS genes. These results confirm the critical role of the MYC2 transcription factor in the blue aleurone trait and provide a biotechnological solution for enriching barley grain with anthocyanins.

Egorova A.A. , Zykova T.E. , Hertig C.W. , Hoffie I. , Morozov S.V. , Chernyak E.I. , Rogachev A.D. , Korotkova A.M. , Vikhorev A.V. , Vasiliev G.V. , Shoeva O.Y. , Kumlehn J. , Gerasimova S.V. , Khlestkina E.K.
Accumulation of Anthocyanin in the Aleurone of Barley Grains by Targeted Restoration of the MYC2 Gene
International Journal of Molecular Sciences. 2024. V.25. N23. 12705 . DOI: 10.3390/ijms252312705 WOS Scopus OpenAlex

Submitted:	Oct 14, 2024
Accepted:	Nov 21, 2024
Published online:	Nov 26, 2024

Web of science:	WOS:001376372900001
Scopus:	2-s2.0-85211921002
OpenAlex:	W4404712325

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