Allelic differences in the key genes of betalain biosynthesis in table beet accessions with contrasting root color from the VIR collection
https://doi.org/10.30901/2227-8834-2024-1-139-151
Abstract
Background. Table beet (Beta vulgaris L.) contains a great amount of naturally red-colored betanins. A number of enterprises are forced to import foreign dyes because the reserves of domestic plant raw materials are insufficient. With this in view, the development of cultivars with high betalain pigment content is now required.
Materials and methods. Allelic differences were screened in the key genes of the betalain biosynthesis pathway among table beet accessions with various root color from the VIR collection using the Sanger DNA sequencing method.
Results and discussion. For the first time we identified a nonsense mutation in the CYP76AD1 gene in cv. ‘Serdolik’ with yellow flesh; it led to the truncation of the functional P450 domain. We suggested that the detected polymorphism correlated with phenotypic switching because the well-known role of CYP76AD1 was essential for the red betalain accumulation. Moreover, a number of missense mutations in cv. ‘Avalanche’ in the first exon of the BvDODA1 gene were found. These mutations were probably associated with the expression of the uncolored phenotype. An in silico analysis revealed highly homologous copies of CYP76AD5, located tandemly on chromosome 9. Attention should be paid to these copies, together with the CYP76AD6 gene, as they seem the most preferable targets for a knockout to increase the red pigment content.
Conclusion. It is possible to identify the best table beet accessions for further genome editing among a previously selected high-betanin group. In addition, this study revealed the allelic differences in the key genes of the betalain biosynthesis pathway. These results will be useful for the development of DNA molecular markers to facilitate the selection of table beet forms with required properties.
Keywords
Beta vulgaris L.,
genes of the cytochrome P450 family,
BvDODA1,
BvMYB1,
betacyanins,
betaxanthins,
root flesh color
Supporting agencies: this research was supported by the Russian Science Foundation under Project No. 21-66-00012 “The development with genetic technologies and the study of new plant lines adapted to changing environmental conditions, with increased productivity and dietary value”. The authors thank the reviewers for their contribution to the peer review of this work.
About the Authors
A. S. Mikhailova
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Aleksandra S. Mikhailova, Associate Researcher,
42, 44 Bolshaya Morskaya Street, St. Petersburg 190000
D. V. Sokolova
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Diana V. Sokolova, Cand. Sci. (Biology), Senior Researcher,
42, 44 Bolshaya Morskaya Street, St. Petersburg 190000
N. A. Shvachko
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Natalia A. Shvachko, Cand. Sci. (Biology), Leading Researcher,
42, 44 Bolshaya Morskaya Street, St. Petersburg 190000
V. S. Popov
N.I. Vavilov All-Russian Institute of Plant Genetic Resources
Russian Federation
Russian Federation
Vitaliy S. Popov, Cand. Sci. (Engineering), Senior Researcher,
42, 44 Bolshaya Morskaya Street, St. Petersburg 190000
E. K. Khlestkina
N.I. Vavilov All-Russian Institute of Plant Genetic Resources;
Sirius University of Science and Technology, Research Center of Genetics and Life Sciences
Russian Federation
Russian Federation
Elena K. Khlestkina, Dr. Sci. (Biology), Professor of the RAS, Director, 42, 44 Bolshaya Morskaya Street, St. Petersburg 190000;
Plant Biology and Biotechnology Research Manager, 1 Olimpiysky Ave., Sirius Settlem., Sirius Federal Territory, Krasnodar Territory 354340
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Supplementary files
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1. Supplement 1 | |
| Subject | Table 1. Allele-specific primers for resequencing alleles of betalain biosynthesis genes in samples contrasting in root color. | |
| Type | Other | |
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(135KB)
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2. Supplement 2 | |
| Subject | Table 2. Allelic differences in structural and regulatory biosynthes betalain genes of table beet. | |
| Type | Other | |
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3. Supplement 3 | |
| Subject | Figure 1. Multiple alignment of nucleotide sequences of the promoter region and the first exon of the CYP76AD1 gene. | |
| Type | Other | |
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4. Supplement 4 | |
| Subject | Figure 2. Multiple alignment of nucleotide sequences of the promoter region and the first exon (highlighted in green) of the CYP76AD5.1 gene. | |
| Type | Other | |
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5. Supplement 5 | |
| Subject | Figure 3. Multiple alignment of nucleotide sequences of the promoter region and the first exon (highlighted with a green frame) of the CYP76AD5.2 gene. | |
| Type | Other | |
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6. Supplement 6 | |
| Subject | Figure 4. Multiple alignment of nucleotide sequences of the promoter region and the first exon (highlighted with a green frame) of the CYP76AD5.3 gene. | |
| Type | Other | |
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7. Supplement 7 | |
| Subject | Figure 5. Multiple alignment of nucleotide sequences of the promoter region and the first exon (highlighted with a green frame) of the CYP76AD6 gene. | |
| Type | Other | |
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8. Supplement 8 | |
| Subject | Figure 6. Multiple alignment of nucleotide sequences of the promoter region and the first exon of the BvDODAl gene. | |
| Type | Other | |
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9. Supplement 9 | |
| Subject | Figure 7. Multiple alignment of nucleotide sequences of the coding region of the BvMYBl gene. | |
| Type | Other | |
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For citations:
Mikhailova A.S., Sokolova D.V., Shvachko N.A., Popov V.S., Khlestkina E.K. Allelic differences in the key genes of betalain biosynthesis in table beet accessions with contrasting root color from the VIR collection. Proceedings on applied botany, genetics and breeding. 2024;185(1):139-151. (In Russ.) https://doi.org/10.30901/2227-8834-2024-1-139-151
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