Investigating of the polymorphism of Solanaceae R2R3 Myb and Brassica Myb114 genes of transcription factors in connection with the anthocyanin biosynthesis regulation

Based on comparison of R2R3 Myb genes in Solanaceae (S. lycopersicum: Ant1, Ant2, S. melongena: Myb1, C. annuum: Myb113-like1 and Myb113-like2) and Brassicaceae crops, a search for orthologous sequences was The sequences encoding Myb114 TF in Brassica oleracea and Brassica rapa was found to be the closest in a nucleotide structure to the previously investigated genes in Solanaceae. Тhе polymorphism in the promoter region of the Capsicum annuum Myb113-like1 gene that regulates anthocyanin biosynthesis: an additional 148 bp repeat and 2(1) bp insertion in the forms with impaired anthocyanin synthesis in fruits was studied. A relationship between the presence of an insert in the promoter (Myb113-like1pr+148) and polymorphisms in the exon regions of Myb113-like1delTand Myb113-like2C/Аgenes associated with impaired anthocyanin synthesis was established. A number of polymorphisms of the Myb114 gene in vegetable crops of the cabbage family (Brassica oleracea, Brassica rapa), which closely correlate with high/low accumulation of anthocyanins in leaves, were identified. In B. oleracea, SNPs that lead to the replacement of two amino acids located in the region of DNAbinding domains were found, which leads to a change in the efficiency of binding of this transcription factor and the promoters of structural biosynthesis genes. A protein sequence encoded by the Myb114 gene in accessions of the leafy turnip (B. rapa) with a high accumulation of anthocyanins in the leaves differed from the accessions without anthocyanin accumulation in the leaves by five amino acids, while the regions of DNA-binding domains were the same in the forms with various anthocyanin accumulation.

1. Middleton E. Jr., Kandaswami C., Theoharides T. C. The effects of plant flavonoids on mammalian cells: Implications for inflammation, heart disease, and cancer. Pharmacological Reviews, 2000, vol. 52, pp. 673–751.

2. Khlestkina E. K., Shoeva O. Yu., Gordeeva E. I. Flavonoid biosynthesis genes in wheat. Vavilovskii zhurnal genetiki i selektsii [Vavilov Journal of Genetics and Breeding], 2014, vol. 18, no. 4/1, pp. 784–796 (in Russian).

3. Liu Y., Tikunov Yu., Schouten R. E., Marcelis L. F. M., Visser R. G. F., Bovy A. Anthocyanin biosynthesis and degradation mechanisms in Solanaceous Vegetables: a review. Frontiers in Chemistry, 2018, vol. 6, pp. 1–17. https://doi.org/10.3389/fchem.2018.00052

4. Naing A. H., Kim C. K. Roles of R2R3-MYB transcription factors in transcriptional regulation of anthocyanin biosynthesis in horticultural plants. Plant Molecular Biology, 2018, vol. 98, no. 1–2, pp. 1–18. https://doi.org/10.1007/s11103-018-0771-4

5. Stommel J. R., Dumm J. M. Coordinated regulation of biosynthetic and regulatory genes coincides with anthocyanin accumulation in developing eggplant fruit. Journal of the American Society for Horticultural Science, 2015, vol. 140, no. 2, pp. 129–135. https://doi.org/10.21273/jashs.140.2.129

6. Lightbourn G. J., Stommel J. R., Griesbach R. J. Epistatic interactions influencing anthocyanin gene expression in Capsicum annuum. Journal of the American Society for Horticultural Science, 2007, vol. 132, no. 6, pp. 824–829. https://doi.org/10.21273/jashs.132.6.824

7. Jung S., Venkatesh J., Kang M.-Y., Kwon J.-K., Kang B.-C. A non-LTR retrotransposon activates anthocyanin biosynthesis by regulating a MYB transcription factor in Capsicum annuum. Plant Science, 2019, vol. 287, art. 110181. https://doi.org/10.1016/j.plantsci.2019.110181

8. Babak O. G., Nekrashevich N. A., Nikitinskaya T. V., Yatsevich K. K., Kilchevsky A. V. Study of the Myb-factor polymorphism based on comparative genomics of vegetable Solanaceae crops (tomato, pepper, eggplant) to search for DNA markers that differentiate samples by the anthocyans accumulation. Doklady Natsional’noi akademii nauk Belarusi = Doklady of the National Academy of Sciences of Belarus, 2019, vol. 63, no. 6, pp. 721–729 (in Russian). https://doi.org/10.29235/15618323-2019-63-6-721-729

9. Babak O., Nikitinskaya T., Nekrashevich N., Yatsevich K., Kilchevsky A. Identification of DNA Markers of Anthocyanin Biosynthesis Disorders Based on the Polymorphism of Anthocyanin 1 Tomato Ortholog Genes in Pepper and Eggplant. Crop Breeding, Genetics and Genomics, 2020, vol. 2, no. 3, art. e200011. https://doi.org/10.20900/cbgg20200011

10. Zhao Z., Xiao L., Xu L., Xing X., Tang G., Du D. Fine mapping the BjPl1 gene for purple leaf color in B2 of Brassica juncea L. through comparative mapping and whole-genome re-sequencing. Euphytica, 2017, vol. 213, no. 4, pp. 80–90. https://doi.org/10.1007/s10681-017-1868-6

11. Rahim M. A., Robin A. H. K., Natarajan S., Jung H.-J., Lee J., Kim H., Kim H.-T., Park J.-I., Nou I.-S. Identification and characterization of anthocyanin biosynthesis-related genes in Kohlrabi. Applied Biochemistry and Biotechnology, 2018, vol. 184, no. 4, pp. 1120–1141. https://doi.org/10.1007/s12010-017-2613-2

12. Wang J., Su T. B., Yu Y. J. Molecular characterization of BrMYB73: a candidate gene for the purple-leaf trait in Brassica rapa. International Journal of Agriculture and Biology, 2019, vol. 22, pp. 122–130. https://doi.org/10.17957/IJAB/15.1041

13. Zhang X., Zhang K., Wu J., Guo N., Liang J., Wang X., Cheng F. QTL-Seq and sequence assembly rapidly mapped the gene BrMYBL2.1 for the purple trait in Brassica rapa. Scientific Reports, 2020, vol. 10, no. 1. https://doi.org/10.1038/s41598020-58916-5

14. Ye J., Coulouris G., Zaretskaya I., Cutcutache I., Rozen S., Madden T. L. Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics, 2012, vol. 13, no. 1, art. 134. https://doi.org/10.1186/1471-2105-13-134

15. Molecular Evolutionary Genetics Analysis. Available at: https://www.megasoftware.net/ (accessed 12 February 2022).

16. Vector NTI. Available at: https://www.thermofisher.com/by/en/home/life-science/cloning/vector-nti-software.html (accessed 5 February 2022).