TSitologiya i Genetika 2023, vol. 57, no. 1, 44-54
Cytology and Genetics 2023, vol. 57, no. 1, 35–43, doi: https://www.doi.org/https://doi.org/10.3103/S0095452723010115

Distribution of alleles of β-carotene hydroxylase 1 gene in modern genotypes of Zea mays L.

Satarova T.M., Denysiuk K.V., Cherchel V.Yu., Dziubetskyi B.V.

  • State Enterprise the Institute of Grain Crops of National Academy of Agrarian Sciences of Ukraine, Volodimir Vernadskij st., 14, Dnipro, 49009, Ukraine

SUMMARY. Carotenoids as precursors for the vitamin A synthesis are important micronutrients for food and feed. β-Carotene is a carotenoid which converts into vitamin A in animal organisms most effectively. The increase in its content in mature maize grain is possible via marker-associated selection by the identification of genotypes with favorable allelic state of the key genes of carotenoid biosynthesis and utilization. The gene of β-carotene hydroxylase 1 is important for the accumulation of β-carotene in mature maize grain. One of its alleles blocks the transition of β-carotene into β-cryptoxanthin and thus ensures the accumulation of β-carotene in mature grain. This allele is detected by the marker crtRB1-3′TE using the polymerase chain reaction as an amplicon of 543 bp unlike two others, 296 bp and 296 + 875 bp, which are not associated with an increase in β-carotene content in the grain of complete maturity. It was established that 26.7 % among 15 well-known inbreds of foreign breeding and 21.6 % of inbreds within 153 perspective inbreds of the Dnipro breeding program carried the allele of β-carotene hydroxylase 1 gene (543 bp) favorable for the accumulation of β-carotene. Allele 543 bp of β-carotene hydroxylase 1 gene by the marker crtRB1-3′TE was found among most of the analyzed maize subspecies, germplasms, and maturity groups. The tendency to its increased frequency for inbreds with flint grain type, Lancaster and Lacon germ-plasms as well as early and middle early inbreds was noticed. Modern perspective inbreds – the carriers of allele 543 bp on gene of β-carotene hydroxylase 1 by marker crtRB1-3′TE, are recommended for the application in special programs of marker-assisted se-lection to increase the content of β-carotene in maize subspecies, groups of maturity, and different types of germplasm.

Keywords: maize, carotenogenesis, β-carotene, molecular genetic markers, inbred, crtRB1 gene

TSitologiya i Genetika
2023, vol. 57, no. 1, 44-54

Current Issue
Cytology and Genetics
2023, vol. 57, no. 1, 35–43,
doi: https://doi.org/10.3103/S0095452723010115

Full text and supplemented materials


Abraham, M.E., Weimer, S.L., Scoles, K., et al., Orange corn diets associated with lower severity of footpad dermatitis in broilers, Poult. Sci., 2021, vol. 100, no. 5, p. 101054. https://doi.org/10.1016/j.psj.2021.101054

Ali, F., Qanmber, G., Li, F., et al., Updated role of ABA in seed maturation, dormancy, and germination, J. Adv. Res., 2022, vol. 35, pp. 199–214. https://doi.org/10.1016/j.jare.2021.03.011

Babu, R., Rojas, N.P., Gao, S., et al., Validation of the effects of molecular marker polymorphisms in LcyE and CrtRB1 on provitamin A concentrations for 26 tropical maize populations, Theor. Appl. Genet., 2013, vol. 2016, pp. 389–399. https://doi.org/10.1007/s00122-012-1987-3

Baseggio, M., Murray, M., Magallanes-Lundback, M., et al., Natural variation for carotenoids in fresh kernels is controlled by uncommon variants in sweet corn, Plant Genome, 2020, vol. 13, no. 1, p. e20008. https://doi.org/10.1002/tpg2.20008

Baveja, A., Muthusamy, V., Panda, K.K., et al., Development of multinutrient-rich biofortified sweet corn hybrids through genomics-assisted selection of shrunken2, opaque2, lcyE and crtRB1 genes, J. Appl. Genet., 2021, https://doi.org/10.1007/s13353-021-00633-4

Carazo, A., Macáková, K., Matoušová, K., et al., Vitamin A update: forms, sources, kinetics, detection, function, deficiency, therapeutic use and toxicity, Nutrients, 2021, vol. 13, no. 5, p. 1703. https://doi.org/10.3390/nu13051703

Chauhan, H.S., Chhabra, R., Rashmi, T., et al., Impact of vte4 and crtRB1 genes on composition of vitamin-E and provitamin-A carotenoids during kernel-stages in sweet corn, J. Food Composit. Analysis, 2022, vol. 105, p. 104264. https://doi.org/10.1016/j.jfca.2021.104264

Cherchel, V.Yu., Dziubetskyi, B.V., Satarova, T.M., et al., Initial Material of Lancaster Germplasm in Maize Selection and Biotechnology, Kyiv: Agrarna Nauka, 2020, pp. 20–43. https://doi.org/10.31073/978-966-540-500-9

Cheremisina, S.H., Grain market in Ukraine: analysis of the current state and development prospects, Ekon. APK, 2021a, no. 2, p. 48. https://doi.org/10.32317/2221-1055.202102048

Cheremisina, S.H., Status and prospects of development of grain exports from Ukraine to African countries, Ekon. APK, 2021b, no. 3, p. 33. https://doi.org/10.32317/2221-1055.202103033

Colombo, R., Ferron, L., and Papetti, A., Colored Corn: An up-date on metabolites extraction, health implication, and potential use, Molecules, 2021, vol. 26, no. 1, p. 199. https://doi.org/10.3390/molecules26010199

Das, A.K., Gowda, M.M., Muthusamy, V., et al., Development of maize hybrids with enhanced vitamin-E, vitamin-A, lysine, and tryptophan through molecular breeding, Front. Plant Sci., 2021, vol. 12, p. 659381. https://doi.org/10.3389/fpls.2021.659381

Duo, H., Hossain, F., Muthusamy, V., et al., Development of sub-tropically adapted diverse provitamin-A rich maize inbreds through marker-assisted pedigree selection, their characterization and utilization in hybrid breeding, PLoS One, 2021, vol. 16, no. 2, p. e0245497. https://doi.org/10.1371/journal.pone.0245497

Goredema-Matongera, N., Ndhlela, T., Magorokosho, C., et al., Multinutrient biofortification of maize (Zea mays L.) in Africa: current status, opportunities and limitations, Nutrients, 2021, vol. 13, no. 3, p. 1039. https://doi.org/10.3390/nu13031039

Goswami, R., Zunjare, R.U., Khan, S., et al., Genetic variability of kernel provitamin-A in sub-tropically adapted maize hybrids possessing rare allele of β-carotene hydroxylase, Cereal Res. Commun., 2019, vol. 47, pp. 205–215. https://doi.org/10.1556/0806.47.2019.12

Graça Dias, M., Borge, G.I.A., Kljak, K., et al., European database of carotenoid levels in foods. Factors affecting carotenoid content, Foods, 2021, vol. 10, no. 5, p. 912. https://doi.org/10.3390/foods10050912

Harjes, C.E., Rocheford, T.R., Bai, L., et al., Natural genetic variation in lycopene epsilon cyclase tapped for maize biofortification, Science, 2008, vol. 319, no. 5861, pp. 330–333. https://doi.org/10.1126/science.1150255

Hwang, T., Ndolo, V.U., Katundu, M., et al., Provitamin A potential of landrace orange maize variety (Zea mays L.) grown in different geographical locations of central Malawi, Food Chem., 2016, vol. 196, pp. 1315–1324. https://doi.org/10.1016/j.foodchem.2015.10.067

Kebede, D., Mengesha, W., Menkir, A., et al., Marker based enrichment of provitamin A content in two tropical maize synthetics, Sci. Rep., 2021, vol. 11, p. 14998. https://doi.org/10.1038/s41598-021-94586-7

Kljak, K., Duvnjak, M., Bedeković, D., et al., Commercial corn hybrids as a single source of dietary carotenoids: effect on egg yolk carotenoid profile and pigmentation, Sustainability, 2021, vol. 13, no. 21, p. 12287. https://doi.org/10.3390/su132112287

Maazou, A.-R.S., Gedil, M., Adetimirin, V.O., et al., Comparative assessment of effectiveness of alternative genotyping assays for characterizing carotenoids accumulation in tropical maize inbred lines, Agronomy, 2021, vol. 11, no. 10, p. 2022. https://doi.org/10.3390/agronomy11102022

Mehta, B.K., Muthusamy, V., Zunjare, R.U., et al., Biofortification of sweet corn hybrids for provitamin-A, lysine and tryptophan using molecular breeding, J. Cereal Sci., 2020, vol. 96, p. 103093. https://doi.org/10.1016/j.jcs.2020.103093

Mehta, B.K., Chhabra, R., Muthusamy, V., et al., Expression analysis of β-carotene hydroxylase1 and opaque2 genes governing accumulation of provitamin-A, lysine and tryptophan during kernel development in biofortified sweet corn, 3 Biotech, 2021, vol. 11, p. 325. https://doi.org/10.1007/s13205-021-02837-1

Menkir, A., Dieng, I., Mengesha, W., et al., Unravelling the effect of provitamin a enrichment on agronomic performance of tropical maize hybrids, Plants (Basel), 2021, vol. 10, no. 8, p. 1580. https://doi.org/10.3390/plants10081580

Mladenović-Drinić, S., Vukadinović, J., Srdić, J., et al., Effect of cooking on the content of carotenoids and tocopherols in sweet corn, Food Feed Res., 2021,vol. 48, no. 2. https://doi.org/10.5937/ffr0-31960

Murray, M.G. and Thompson, W.F., Rapid isolation of high molecular weight plant DNA, Nucleic Acids Res., 1980, vol. 8, pp. 4321–4325.

Muthusamy, V., Hossain, F., Thirunavukkarasu, N., et al., Allelic variations for lycopene-ε-cyclase and β-carotene hydroxylase genes in maize inbreds and their utilization in β-carotene enrichment programme, Cogent Food Agric., 2015, vol. 1, no. 1, p. 1033141. https://doi.org/10.1080/23311932.2015.1033141

Naqvi, Sh., Zhu, Ch., Farre, G., et al., Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways, Proc. Natl. Acad. Sci. U. S. A., 2009, vol. 106, no. 19, p. 7762–7767. https://doi.org/10.1073/pnas.0901412106

Natesan, S., Singh, T.S., Duraisamy, T., et al., Characterization of crtRB1 gene polymorphism and β-carotene content in maize landraces originated from North Eastern Himalayan region (NEHR) of India, Front. Sustainable Food Syst., 2020, vol. 4, p. 78. https://doi.org/10.3389/fsufs.2020.00078

Petrychenko, V.F. and Tomashuk, O.V., Features of formation of indicators of quality of corn grain at various technologies of cultivation in the Foreststeppe of the Right bank, Plant Soil Sci., 2019, vol. 10, no. 2. https://doi.org/10.31548/agr2019.02.029

Qutub, M., Chandran, S., Rathinavel, K., et al., Improvement of a Yairipok Chujak maize landrace from North Eastern Himalayan region for β-carotene content through molecular marker-assisted backcross breeding, Genes, 2021, vol. 12, no. 5, p. 762. https://doi.org/10.3390/genes12050762

Roca, M. and Pérez-Gálvez, A., Metabolomics of chlorophylls and carotenoids: analytical methods and metabolome-based studies, Antioxidants (Basel), 2021, vol. 10, no. 10, p. 1622. https://doi.org/10.3390/antiox10101622

Saenz, E., Borrás, L., and Gerde, J.A., Carotenoid profiles in maize genotypes with contrasting kernel hardness, J. Cereal Sci., 2021, vol. 99, p. 103206. https://doi.org/10.1016/j.jcs.2021.103206

Sagare, D., Shetti, P., Surender, M., et al., Marker-assisted backcross breeding for enhancing β-carotene of QPM inbreds, Mol. Breed., 2019, vol. 39, no. 2, p. 31. https://doi.org/10.1007/s11032-019-0939-x

Satarova, T.M., Semenova, V.V., Zhang, J., et al., Differentiation of maize breeding samples by β-carotene content, Regul. Mech. Biosyst., 2019, vol. 10, no. 1, pp. 63–68. https://doi.org/10.15421/021910

Sathasivam, R., Radhakrishnan, R., Kim, J.K., et al., An update on biosynthesis and regulation of carotenoids in plants, S. Afr. J. Bot., 2021, vol. 140, pp. 290–302. https://doi.org/10.1016/j.sajb.2020.05.015

Sun, X., Ma, L., Lux, P.E., et al., The distribution of phosphorus, carotenoids and tocochromanols in grains of four Chinese maize (Zea mays L.) varieties, Food Chem., 2022, vol. 367, p. 130725. https://doi.org/10.1016/j.foodchem.2021.130725

Von Lintig, J., Moon, J., and Babino, D., Molecular components affecting ocular carotenoid and retinoid homeostasis, Prog. Retinal Eye Res., 2020, vol. 80, p. 100864. https://doi.org/10.1016/j.preteyeres.2020.100864

Welham, S.J., Gezan, S.A., Clark, S.J., et al., Statistical Methods in Biology: Design and Analysis of Experiments and Regression, Boca Raton: CRC Press, 2014.


Yan, J., Kandianis, C.B., Harjes, C.E., et al., Rare genetic variation at Zea mays crtRB1 increases betacarotene in maize grain, Nat. Genet., 2010, vol. 42, pp. 322–327. https://doi.org/10.1038/ng.551

Zafar, J., Aqeel, A., Shah, F.I., et al., Biochemical and immunological implications of Lutein and Zeaxanthin, Int. J. Mol. Sci., 2021, vol. 22, no. 20, p. 10910. https://doi.org/10.3390/ijms222010910

Zatyshniak, O.V., Cherchel, V.Yu., Dziubetskyi, B.V., et al., Marker-assisted selection for the gene of β-carotene hydroxylase in maize, Factory Exp. Evol. Org., 2020, vol. 27, pp. 83–88. https://doi.org/10.7124/FEEO.v27.1307

Zhai, S.N., Xia, X.C., and He, óL.H., Carotenoids in staple cereals: metabolism, regulation and genetic manipulation, Front. Plant Sci., 2016, vol. 7, p. 1197. https://doi.org/10.3389/fpls.2016.01197

Zurak, D., Grbeša, D., Duvnjak, M., et al., Carotenoid content and bioaccessibility in commercial maize hybrids, Agriculture, 2021, vol. 11, no. 7, p. 586. https://doi.org/10.3390/agriculture11070586