Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil

Povkhova, Liubov V.; Pushkova, Elena N.; Rozhmina, Tatiana A.; Zhuchenko, Alexander A.; Frykin, Roman I.; Novakovskiy, Roman O.; Dvorianinova, Ekaterina M.; Gryzunov, Aleksey A.; Borkhert, Elena V.; Sigova, Elizaveta A.; Vladimirov, Gleb N.; Snezhkina, Anastasiya V.; Kudryavtseva, Anna V.; Krasnov, George S.; Dmitriev, Alexey A.; Melnikova, Nataliya V.

doi:10.3390/plants12010095

Open AccessArticle

Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil

by

Liubov V. Povkhova

¹

,

Elena N. Pushkova

¹,

Tatiana A. Rozhmina

^1,2,

Alexander A. Zhuchenko

^2,3,

Roman I. Frykin

^1,4,

Roman O. Novakovskiy

¹,

Ekaterina M. Dvorianinova

^1,5

,

Aleksey A. Gryzunov

⁶,

Elena V. Borkhert

¹,

Elizaveta A. Sigova

^1,5,

Gleb N. Vladimirov

⁷,

Anastasiya V. Snezhkina

¹

,

Anna V. Kudryavtseva

¹

,

George S. Krasnov

¹

,

Alexey A. Dmitriev

¹

and

Nataliya V. Melnikova

^1,*

¹

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia

²

Federal Research Center for Bast Fiber Crops, 172002 Torzhok, Russia

³

All-Russian Horticultural Institute for Breeding, Agrotechnology and Nursery, 115598 Moscow, Russia

⁴

Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia

⁵

Moscow Institute of Physics and Technology, 141701 Moscow, Russia

⁶

All-Russian Scientific Research Institute of Refrigeration Industry—Branch of V.M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences, 127422 Moscow, Russia

⁷

Skolkovo Institute of Science and Technology, 121205 Moscow, Russia

^*

Author to whom correspondence should be addressed.

Plants 2023, 12(1), 95; https://doi.org/10.3390/plants12010095

Submission received: 11 October 2022 / Revised: 14 December 2022 / Accepted: 16 December 2022 / Published: 24 December 2022

(This article belongs to the Special Issue Plant Molecular Breeding)

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Abstract

Flax is grown worldwide for seed and fiber production. Linseed varieties differ in their oil composition and are used in pharmaceutical, food, feed, and industrial production. The field of application primarily depends on the content of linolenic (LIN) and linoleic (LIO) fatty acids. Inactivating mutations in the FAD3A and FAD3B genes lead to a decrease in the LIN content and an increase in the LIO content. For the identification of the three most common low-LIN mutations in flax varieties (G-to-A in exon 1 of FAD3A substituting tryptophan with a stop codon, C-to-T in exon 5 of FAD3A leading to arginine to a stop codon substitution, and C-to-T in exon 2 of FAD3B resulting in histidine to tyrosine substitution), three approaches were proposed: (1) targeted deep sequencing, (2) high resolution melting (HRM) analysis, (3) cleaved amplified polymorphic sequences (CAPS) markers. They were tested on more than a thousand flax samples of various types and showed promising results. The proposed approaches can be used in marker-assisted selection to choose parent pairs for crosses, separate heterogeneous varieties into biotypes, and select genotypes with desired homozygous alleles of the FAD3A and FAD3B genes at the early stages of breeding for the effective development of varieties with a particular LIN and LIO content, as well as in basic studies of the molecular mechanisms of fatty acid synthesis in flax seeds to select genotypes adequate to the tasks.

Keywords: flax; linseed; fatty acid composition; linolenic acid; FAD3 genes; DNA markers; marker-assisted selection; targeted deep sequencing; HRM; CAPS

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MDPI and ACS Style

Povkhova, L.V.; Pushkova, E.N.; Rozhmina, T.A.; Zhuchenko, A.A.; Frykin, R.I.; Novakovskiy, R.O.; Dvorianinova, E.M.; Gryzunov, A.A.; Borkhert, E.V.; Sigova, E.A.; et al. Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil. Plants 2023, 12, 95. https://doi.org/10.3390/plants12010095

AMA Style

Povkhova LV, Pushkova EN, Rozhmina TA, Zhuchenko AA, Frykin RI, Novakovskiy RO, Dvorianinova EM, Gryzunov AA, Borkhert EV, Sigova EA, et al. Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil. Plants. 2023; 12(1):95. https://doi.org/10.3390/plants12010095

Chicago/Turabian Style

Povkhova, Liubov V., Elena N. Pushkova, Tatiana A. Rozhmina, Alexander A. Zhuchenko, Roman I. Frykin, Roman O. Novakovskiy, Ekaterina M. Dvorianinova, Aleksey A. Gryzunov, Elena V. Borkhert, Elizaveta A. Sigova, and et al. 2023. "Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil" Plants 12, no. 1: 95. https://doi.org/10.3390/plants12010095

APA Style

Povkhova, L. V., Pushkova, E. N., Rozhmina, T. A., Zhuchenko, A. A., Frykin, R. I., Novakovskiy, R. O., Dvorianinova, E. M., Gryzunov, A. A., Borkhert, E. V., Sigova, E. A., Vladimirov, G. N., Snezhkina, A. V., Kudryavtseva, A. V., Krasnov, G. S., Dmitriev, A. A., & Melnikova, N. V. (2023). Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil. Plants, 12(1), 95. https://doi.org/10.3390/plants12010095

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Development and Complex Application of Methods for the Identification of Mutations in the FAD3A and FAD3B Genes Resulting in the Reduced Content of Linolenic Acid in Flax Oil

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