Development of high-lycopene tomato hybrids using conventional breeding techniques and molecular markers

Relevance. High lycopene fruit content has been regarded as a very important genetic trait in tomato breeding. Use lycopene molecular markers in combination with conventional breeding techniques allowed us to create hybrids with high lycopene accumulation, excellent organoleptic qualities, high yield production and resistance to pathogens, and to effectively optimize our breeding programmes for commercial greehouses production.

Material and Methods. In this study tomato samples including selected lines and hybrids with various allelic combinations of genes determining carotene accumulation, and other genetic traits, such as disease resistance and yield production were tested. Introgression of spontaneous and induced mutations was used to increase carotenoid levels (og and hp) and improve fruit technological qualities (nor, alc, rin). The research material was tomato collection, mutants, breeding lines and hybrids listed in the State Register Russian Federation tomato hybrids of breeding SS Agrofirm "Ilyinichna" VNIIO branch of the All-Russian Scientific Research Institute of Vegetable Growing – Branch of the FSBSI Federal Scientific Vegetable Center. DNA typing of fruit quality genes was performed at the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus.

Results. New domestic hybrids for industrial greenhouses, which characterised by improved organoleptic qualities and technological traits were developed with the help of phasedcross-breeding that allowed to combine the genes nor, rin, alc, leading to an extension of the shelf life with the genes B, og, hp1, etc., contributing to an increase in carotenoid content in fruits. It was established that for targeted selection and hybridization, despite the negative influence of the nor, rin, alc genes it is possible to raise the level of carotenoids to average values. Correlation between lycopene concentration in fruits and high temperature and level of insolation was confirmed. It was shown that pink-fruited forms contain significantly more lycopenethanred-fruitedones. Different all eliccombinations of structural genes involved in carotenoids biosynthesis and regulatory genes that provided maximal accumulation of lycopene in hybrid swithred and pink fruits were revealed. Hybrids with the combination of high concentrations of sugar (° Brix), dry matter and maximal lycopene values, combined defining excellent taste were selected: Prekrasnaiya lady, Olya, Quadrille, Victoria. New F₁ hybrids one for industrial greenhouses: G950, G956, G960, Magistral and pink fruited G12897, surpassed the Dutch standard in productivity up to 21%, and in tastes/organoleptic qualities for 1-1.8 points.

1. Tan H.L. et al. Tomato-based food products for prostate cancer prevention: what have we learned? Cancer Metastasis Rev. 2010;29(3):553-568. doi: 10.1007/s10555-010-9246-z.;

2. Sporn M.B., Liby K.T. Is lycopene an effective agent for preventing prostate cancer? Cancer Prev. Res. 2013;6(5):384-386. doi: 10.1158/1940-6207.CAPR13-0026.

3. Nisar N., Li Li, Lu S., Khin N.C., Pogson B.J. Carotenoid metabolism in plants. Mol Plant. 2015 Jan;8(1):68-82. doi: 10.1016/j.molp.2014.12.007.

4. Sun, T., Yuan, H., Cao, H., Yazdani, M., Tadmor, Y., and Li, L. Carotenoid metabolism in plants: the role of plastids. Mol. Plant. 2018;(11):58–74. doi: 10.1016/j.molp.2017.09.010

5. Ilan Levin, C.H. Ric de Vos, Yaakov Tadmor, Arnaud Bovy, Michal Lieberman, Michal OrenShamir, Orit Segev, Igor Kolotilin, Menachem Keller, Rinat Ovadia, Ayala Meir, and Raoul J. Bino High pigment tomato mutants— more than just lycopene (a review). Israel Journal of Plant Sciences. 2006;(54):179–190.

6. Labate J.A., Grandillo S., Fulton T., Munos S. Tomato. Genome mapping and molecular breeding in plants. Vol. 5. Vegetables, ed. C. Kole. Berlin, Heidelberg, New York: Springer, 2007:1–126.

7. GenBank. Available at: (accessed 1 Aug 2019).

8. Solgenomics. Available at: https://solgenomics.net/ (accessed 1 Aug 2019).

9. Kilchevsky A.V., Babak O.G., Malyshev S.V., Adzhieva V.F., Nekrashevich N.A., Yatsevich K.K., Kondratyuk A.V. DNA typing of genes for fruit quality and resistance to tomato diseases. Guidelines. The Ministry of Agriculture and Food of the Republic of Belarus, the National Academy of Sciences of Belarus, the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus. Minsk, 2016. 41 р. (in Russian)

10. Babak O.G., Nekrashevich N.A., Yatsevich K.K., Мalyshev S.V., Kilchevsky A. V. Genetic bases of tomatо marker-assisted selection in Belarus. Eurobiotech. J. 2018;2(2):128-135, doi:10.2478/ebtj-2018-0017

11. Ronen, G., Carmel-Goren L. , Zamir D. Hirschberg J. An alternative pathway to β-carotene formation in plant chromoplasts discovered by mapbased cloning of Beta and old-gold color mutations in tomato. PNAS. 2000;97(20):11102-11107.

12. Vrebalov, J., Ruezinsky D., Padmanabhan V., White R., Medrano D., Drake R., Schuch W., Giovannoni J. A MADS-box gene necessary for fruit ripening at the tomato ripening-inhibitor (Rin) locus. Science. 2002;(296):343–345.

13. Tigchelaar, E.C., M.L. Tomes, E.A. Kerr, and R.J. Barman. A new fruit ripening mutant, nonripening (nor). Rep. Tomato Genet. Coop. 1973. P.23-33.

14. Kolotilin, I., Koltai H., Tadmor Y., Bar-Or C., Reuveni M., Meir A., Nahon S., Shlomo H., Chen L., Levin I. Transcriptional profiling of high pigment-2dg tomato mutant links early fruit plastid biogenesis with its overproduction of phytonutrients. Plant Physiology. 2007;(145):389–401.

15. Yen, H.C., Shelton B.A., Howard L.R., Lee S. , Vrebalov J., Giovannoni J. J. The tomato highpigment (hp) locus maps to chromosome 2 and influences plastome copy number and fruit quality. Theor Appl Genet.1997;(95):1069–1079.

16. Kim B., Kim N., Kang J., Choi Y., Sim S.-C., Min S.R., Park Y. Single Nucleotide Polymorphisms linked to the SlMYB12 Gene that Controls Fruit Peel Color in Domesticated Tomatoes (Solanum lycopersicum L.). Kor. J. Hort. Sci. Technol. 2015;33(4):566-574.

17. Veerappan K., Jung H.J., Hwang I., Kho K.H., Chung M.Y., Nou I.S. Sequence Variation in SlMYB12 is Associated with Fruit Peel Color in Pink Tomato Cultivar. Hortic. Environ. Biotechnol. 2016;57(3):274-279.

18. Golubkina N.A., Mоlchanova A.V., Tareeva M.M., Baback O.G., Nekrashevich N.A., Kondratyeva I.Y. Quantitative thing layer chromatography for evaluation of carotenoid composition of tomatoes Solanum lycopersicum. Vegetable crops of Russia. 2017;(5):96-99. (In Russ.) https://doi.org/10.18619/2072-91462017-5-96-99

19. Guidelines for the selection of tomato varieties and hybrids for open and protected soil. 1986. 98 p. (in Russian)

20. Methods for the selection of tomato for resistance to powdery mildew (Oidium lycopersicum Cooke et Massee). 2005. 28 p. (in Russian)

21. Kuzeminsky A.V. Selection and genetic studies of mutant forms of tomato. Kharkov, 2004. 392 р. (in Russian)

22. Bagirova S.F., Ignatova S.I. Molecular methods in plant breeding. Gavrish. 2012;2:33-38 (in Russian)

23. Bramley P. Regulation of carotenoid formation during tomato fruit ripening and development (Review). Journal of Experimental Botany. 2002;53(377):2107-13 • November 2002, DOI: 10.1093/jxb/erf059

24. Ilahy R., Imen T., Siddiqui M.W., Montefusco A. When color really matters: horticultural performance and functional quality of high-lycopene tomatoes. Plant Sciences. 2018, DOI: 10.1080/07352689.2018.1465631