Use of molecular markers associated with resistance to biotic and abiotic environmental factors in developing breeding material for tomato and pepper in Belarus

Relevance. The development of a system of molecular markers that allows identifying the genetic determinants of resistance to pathogens, as well as the typing of alleles involved in the regulation of anthocyanin accumulation, is the most important condition for increasing the efficiency of the breeding process aimed at enhancing the resistance of cultivated crops to biotic and abiotic stresses.

Methodology. The work involved molecular genetic methods of DNA isolation, PCR analysis, restriction, and evaluation of amplification and restriction products in agarose or polyacrylamide gels. The material used included the diverse collections of Solanum lycopersicum and Capsicum annuum, as well as the specimens of closely related wild species.

Results. The paper evaluates the effectiveness of 25 molecular markers presented in the literature associated with resistance to tomato and pepper diseases caused by fungal, bacterial, viral pathogens, as well as nematodes. Markers to the alleles of MYB transcription factor genes associated with the regulation of anthocyanin accumulation in tomato (SlMyb12, Anthocyanin1, Anthocyanin2, An-2-like and Atroviolacium) and pepper (Myb113-like1, Myb113-like2 and ETC3-2), recommended for the breeding process aimed at increasing resistance to stressful biotic and abiotic environmental factors, are presented.

1. Biotechnology and plant disease management / ed. by Z.K. Punja, S.H. De Boer, H. Sanfaçon / Biddles Ltd, King’s Lynn, UK. 2007. 574 p.

2. Lee J.M., Oh C., Yeam I. Molecular Markers for Selecting Diverse Disease Resistances in Tomato Breeding Programs. Plant Breeding and Biotechnology. 2015;(3):308-322. https://doi.org/10.9787/PBB.2015.3.4.308

3. Truong H.T.H., Choi H., Cho M.C., Lee H.E., Kim J.H. Use of Cf-9 gene-based markers in marker-assisted selection to screen tomato cultivars with resistance to Cladosporium fulv um. Hortic. Environ. Biotechnol. 2011;(52):204–210. https://doi.org/10.1007/s13580-011-0164-y

4. Kang W., Hoang N., Yang H., Kwon J., Jo S., Seo J., Kim K., Choi D., Kang B. Molecular mapping and characterization of a single dominant gene controlling CMV resistance in peppers (Capsicum annuum L.). Theor. Appl. Genet. 2010;(120):1587–1596. https://doi.org/10.1007/s00122-010-1278-9

5. Uncu A.T., Celik I., Devran Z., Ozkaynak E., Frary A., Frary A., Doganla S. Development of SNP-based CAPS assay for the Me1 gene conferring resistance to root knot nematode in pepper. Euphytica. 2015;(206):393–399. https://doi.org/10.1007/s10681-015-1489-x

6. Kim H., Nahm S., Lee H., Yoon G., Kim K., Kang B., Choi D., Kweon O.Y., Cho M., Kwon J., Han J., Kim J., Park M., Ahn J.H., Choi S.H., Her N.H., Sung J., Kim B. BAC-derived markers converted from RFLP linked to Phytophthora capsici resistance in pepper (Capsicum annuum L. Theor. Appl. Genet. 2008;(118):15–27.

7. ChunYing S., Li M., Hai Z., Alainb P., Haoa W., Xi Z. Resistances to anthracnose (Colletotrichum acutatum) of Capsicum mature green and ripe fruit are controlled by a major dominant cluster of QTLs on chromosome P5. Scientia Horticulturae. 2015;(181):81-88. https://doi.org/10.1016/j.scienta.2014.10.033

8. Paran I., Benarous S., Ashkenazi V. Disease resistant pepper plants. Hazera Genetics Ltd, The Agricultural Research Organization. Volcani Center, 2009. Patent WO2009098685 A2

9. Lee H.R., An H.J., You Y.G., Lee J., Kim H.J., Kang B.C., Harn C.H. Development of a novel codominant molecular marker for Chili veinal mottle virus resistance in Capsicum annuum L. Euphytica. 2013;(193):197–205.

10. Quirin E.A., Ogundiwin E.A., Prince J.P., Mazourek M., Briggs M.O., Chlanda T.S., Kim K., Falise M., Kang B., Jahn M.M. Development of sequence characterized amplified region (SCAR) primers for the detection of Phyto.5.2, a major QTL for resistance to Phytophthora capsici Leon. in pepper. Theor. Appl. Genet. 2005;(110):605–612. https://doi.org/10.1007/s00122-004-1874-7

11. Özkaynak E., Devran Z., Kahveci E., Doğanlar S., Başköylü B., Doğan F., İşleyen M., Yüksel A., Yüksel M. Pyramiding Multiple Genes for Resistance to PVY, TSWV and PMMoV in Pepper Using Molecular Markers. Europ. J. Hort. Sci. 2014;79(4):233–239.

12. Rӧmer P., Jordan T., Lahaye T. Identification and application of a DNA-based marker that is diagnostic for the pepper (Capsicum annuum) bacterial spot resistance gene Bs3. Plant Breed. 2010;(129):737–740. https://doi.org/10.1111/j.1439-0523.2009.01750.x

13. Babak O.G., Nekrashevich N.A., Anisimova N.V., Drozd E.V., Yatsevich K.K., Kilchevsky A.V. Technology of marker-assisted selection of tomato forms with high biochemical and technological properties of fruits: methodological recommendations. Minsk: Ministry of Agriculture and Food of the Republic of Belarus; National Academy of Sciences of Belarus; Institute of Genetics and Cytology of the National Academy of Sciences of Belarus Law and Economics, 2023. 74 p. (In Russ.)

14. Ignatova S.I., Babak O.G., Bagirova S.F. Development of high-lycopene tomato hybrids using conventional breeding techniques and molecular markers. Vegetable crops of Russia. 2020;(5):22-28. (In Russ.) https://doi.org/10.18619/2072-9146-2020-5-22-28 https://elibrary.ru/avqfuj (In Russ.)

15. Babak O.G., Drozd E.V., Nekrashevich N.A., Anisimova N.V., Yatsevich K.K., Bayeva I.E., Pugachova I.G., Frantsuzionak A.V., Dobrodkin M.M., Kilchevsky A.V.Assessment and application of molecular markers in breeding for the resistance of tomato (Solanum lycopersicum L.) to late blight (Phytophthora infestans). Molecular and Applied Genetics. 2021;(31):22-30. https://doi.org/10.47612/1999-9127-2021-31-22-30 https://elibrary.ru/kgdhdp (In Russ.)

16. Liu Y., Tikunov Y., Schouten R.E., Marcelis L., Visser R., Bovy A. Anthocyanin biosynthesis and degradation mechanisms in Solanaceous Vegetables: a review. Frontiers in Chemistry. 2018;6(52):1–17. https://doi.org/10.3389/fchem.2018.00052

17. Bassolino L., Zhang Y., Schoonbeek H.J., Kiferle C., Perata P., Martin C. Accumulation of anthocyanins in tomato skin extends shelf life. New Phytol. 2013;200(3):650-655. https://doi.org/10.1111/nph.12524

18. Zhang Y., Butelli E., De Stefano R., Schoonbeek H.J., Magusin A., Pagliarani C., Wellner N., Hill L., Orzaez D., Granell A., Jones J.D., Martin C. Anthocyanins double the shelf life of tomatoes by delaying overripening and reducing susceptibility to gray mold. Curr. Biol. 2013;23(12):1094-100. https://doi.org/10.1016/j.cub.2013.04.072

19. Sapir M., Oren-Shamir M., Ovadia R., Reuveni M., Evenor D., Tadmor Y., Nahon S., Shlomo H., Chen L., Meir A., Levin I. Molecular Aspects of Anthocyanin fruit Tomato in Relation to high pigment-1. Journal of Heredity. 2008;99(3):292–303. https://doi.org/10.1093/jhered/esm128

20. Cao X., Qiu Z., Wang X., Van Giang T., Liu X., Wang J., Wang X., Gao J., Guo Y., Du Y., Wang G., Huang Z. A putative R3 MYB repressor is the candidate gene underlying atroviolacium, a locus for anthocyanin pigmentation in tomato fruit. J. Exp. Bot. 2017;68(21-22):5745-5758. https://doi.org/10.1093/jxb/erx382

21. Babak O.G., Drozd E.V., Nekrashevich N.A., Anisimova N.V., Yatsevich K.K., Kilchevsky A.V. Development of molecular markers for the accumulation of anthocyanins in fruits and studying the specifics of Ant1, An2 and Atv gene interaction in Solanum lycopersicum. Molecular and Applied Genetics. 2024;(36):7-23. https://elibrary.ru/gdbfhm (In Russ.)

22. 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 of the National Academy of Sciences of Belarus. 2019;63(6):721-729. – https://doi.org/10.29235/1561-8323-2019-63-6-721-729 https://elibrary.ru/bmbsqh (In Russ.)

23. 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 Breed Genet Genom. 2020;2(3):e200011. https://doi.org/10.20900/cbgg20200011

24. Babak O.G., Anisimova N.A., Nikitinskaya T.V., Nekrashevich N.A., Yatsevich K.K., Drozd L.V., Fateev D.A., Berensen F.A., Artemyeva A.M., Kilchevsky A.V.Investigating of the polymorphism of Solanaceae R2R3 Myb and Brassica Myb114 genes of transcription factors in connection with the anthocyanin biosynthesis regulation. Doklady of the National Academy of Sciences of Belarus. 2022;66(4):414-424. https://doi.org/10.29235/1561-8323-2022-66-4-414-424 https://elibrary.ru/fuwcwf (In Russ.)