EFFECTIVE METHOD FOR GENETIC TRANSFORMATION OF POTATOES AND CREATION OF IN VITRO COLLECTION OF TRANSGENIC PLANTS WITH GENES THAT INCREASE RESISTANCE TO ABIOTIC AND BIOTIC STRESSES
DOI:
https://doi.org/10.15407/dopovidi2026.01.050Keywords:
genetic engineering, Agrobacterium tumefaciens, potato (Solanum tuberosum), heterologous genesAbstract
Currently, various agricultural crops are exposed to multiple environmental stresses, including salinity, drought, extreme temperatures, mineral deficiencies or excesses, etc. Potatoes (Solanum tuberosum) are an important food, feed, and industrial crop in Ukraine and worldwide. Polyploidy in cultivated Solanum tuberosum is a serious obstacle to its breeding. Genetic engineering has a number of advantages over traditional breeding in terms of increasing the crop’s resistance to stress and allows for the rapid creation of resistant plants based on existing commercial varieties. Sustainable production strategies require the optimization of existing protocols for potato genetic transformation and the transfer of stress-resistant genes, which remain relevant and were the focus of our work. Genes of two isoforms of the vacuolar Na+/H+-antiporter from barley (HvNHX2 and HvNHX3) contribute to increased resistance to drought, salinity and soil alkalinity. A short antisense segment of the proline dehydrogenase (PDH ex1) gene from Arabidopsis thaliana reduces the activity of this enzyme by partially suppressing the PDH gene, thereby increasing proline content and resistance of transgenic plants to drought, cold, heavy metals and salinity. The cytochrome P450 gene from the bovine adrenal cortex (CYP11A1) increases yield and resistance to phytopathogenic fungi. Genes for extracellular RNase from Zinnia (ZRNase) and bovine pancreatic RNase (bov) increase plant resistance to viral infections. Commercial potato varieties were transformed via Agrobacterium tumefacience route using vectors containing the above-mentioned genes. The resulting collection of transgenic potato lines represents valuable material for future investigation on stress resistance and allows comparison of the influence of different alien genes across the certain backgrounds of a number of commercial varieties.
Downloads
References
Vinterhalter, D., Zdravković-Korać, S., Mitić, N., Dragićević, I., Cingel, A., Raspor, M. & Ninković, S. (2008). Protocols for Agrobacterium-mediated transformation of potato. Fruit, Veg. Cereal Sci. Biotech., 2, Spec. Iss. 1: Potato I, pp. 1-15. Retrieved from https://www.researchgate.net/publication/328732619_Protocols_for_ Agrobacterium_mediated_transformation_of_potato
del Mar Martínez-Prada, M., Curtin, S. J. & Gutiérrez-González, J. J. (2021). Potato improvement through genetic engineering. GM Crops Food, 12, No. 1, pp. 479-496. https://doi.org/10.1080/21645698.2021.1993688
Rather, G. A., Ayzenshtat, D., Teper-Bamnolker, P., Kumar, M., Forotan, Z., Eshel, D. & Bocobza, S. (2022). Advances in protoplast transfection promote efficient CRISPR/Cas9-mediated genome editing in tetraploid potato. Planta, 256, No. 1, 14. https://doi.org/10.1007/s00425-022-03933-z
Kikuchi, A., Huynh, H. D., Endo, T. & Watanabe, K. (2015). Review of recent transgenic studies on abiotic stress tolerance and future molecular breeding in potato. Breed. Sci., 65, No. 1, pp. 85-102. https://doi.org/10.1270/jsbbs.65.85
Sakhno, L. O., Gerasymenko, I. M., Komarnitsskii, I. K., Sheludko, Y. V. & Goldenkova-Pavlova, I. V. (2012). Creation of glyphosate-resistant Brassica napus L. plants expressing DesC desaturase of cyanobacterium Synechococcus vulcanus. Biopolym. Cell, 28, No. 6, pp. 449-455. https://doi.org/10.7124/bc.000135
Sakhno, L. O., Slyvets, M. S. & Kuchuk, M. V. (2014) cyp11A1 Canola plants under short time heat stress conditions. Cytol. Genet., 48, pp. 279-284. https://doi.org/10.3103/S0095452714050090
Tishchenko, O. M., Komisarenko, A. G., Mykhalska, S. I., Sergeeva, L. E., Adamenko, N. I., Morgun, B. V. & Kochetov, A. V. (2014). Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.) in vitro and in planta using LBA4404 strain harboring binary vector pBi2E with dsRNA-suppressor of proline dehydrogenase gene. Cytol. Genet., 48, No. 4, pp. 218-226. https://doi.org/10.3103/S0095452714040094
Potrokhov, A., Sosnovska, D., Ovcharenko, O., Budzanivska, I., Rudas, V. & Kuchuk, M. (2021). Increased ribonuclease activity in Solanum tuberosum L. transformed with heterologous genes of apoplastic ribonucleases as a putative approach for production of virus resistant plants. Turk. J. Biol., 45, No. 1, pp. 79-87. https://doi. org/10.3906/biy-2007-87
Potrokhov, A. O. & Ovcharenko, O. O. (2024). Strategies for engineering of virus-resistant plants: focus on RNases. Cytol. Genet., 58, pp. 99-114. https://doi.org/10.3103/S0095452724020099
Ovcharenko, O., Zholobak, N., Rudas, V. & Kuchuk, M. (2025). Plant systems as platforms for the production ofinterferonalphaanditsapplication.Cytol.Genet.,59,pp.622-633.https://doi.org/10.3103/S0095452725060088
Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant., 15, No. 3, pp. 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052
Bayat, F., Shiran, B. & Belyaev, D. V. (2011). Overexpression of HvNHX2, a vacuolar Na+/H+ antiporter gene
from barley, improves salt tolerance in Arabidopsis thaliana. Aust. J. Crop Sci., 5, No. 4, pp. 428-432. Retrieved from https://www.cropj.com/bayat_5_4_2011_428_432.pdf
Krivosheeva, A. B., Varlamova, T. V., Yurieva, N. O., Sobol’kova, G. I., Kholodova, V. P. & Belyaev, D. V. (2014). Potato transformation with the HvNHX3 gene and the improvement of transformant salt tolerance. Russ. J. Plant Physiol., 61, pp. 792-800. https://doi.org/10.1134/S1021443714060119
Trifonova, E. A., Sapotsky, M. V., Komarova, M. L., Scherban A. B., Shumny V. K., Polyakova A. M., Lapshi- na L. A., Kochetov A. V. & Malinovsky V. I. (2007). Protection of transgenic tobacco plants expressing bovine pancreatic ribonuclease against tobacco mosaic virus. Plant Cell Rep., 26, No. 7, pp. 1121-1126. https://doi. org/10.1007/s00299-006-0298-z
Trifonova, E. A., Romanova, A. V., Sangaev, S. S., Sapotsky, M. V., Malinovsky, V. I. & Kochetov, A. V. (2012). Inducible expression of the gene of Zinnia elegans coding for extracellular ribonuclease in Nicotiana tabacum plants. Biol. Plant., 56, pp. 571-574. https://doi.org/10.1007/s10535-011-0206-4
Berdichevets, I. N., Shimshilashvili, H. R., Gerasymenko, I. M., Sindarovska, Y. R., Sheludko, Y. V. & Goldenkova- Pavlova, I. V. (2010). Multiplex PCR assay for detection of recombinant genes encoding fatty acid desaturases fused with lichenase reporter protein in GM plants. Anal. Bioanal. Chem., 397, No. 6, pp. 2289-2293. https:// doi.org/10.1007/s00216-010-3770-0
Bertani, G. (1951). Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol., 62, No. 3, pp. 293-300. https://doi.org/10.1128/jb.62.3.293-300.1951
Doyle, J. J. & Doyle, J. L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12, No. 1, pp. 13-15.
Rudas, V. A., Shakhovsky, A. M., Morgun, B. V., Matveeva, N. A. & Kuchuk, M. V. (2009). Obtaining of transgenic potato plants resistant to the herbicide basta, containing the gene cyp11A1 of cytochrome P450scc. Faktori eksperimental'noï evolûcìï organìzmìv, 7, pp. 192-196 (in Ukrainian).
Kyrpa-Nesmiyan, Т. М. (2017). Study of heterological expression of cyanobacterial desaturase genes in higher plants (Unpublished candidate thesis). Institute of Cell Biology and Genetic Engineering of the NAS of Ukraine, Kyiv, Ukraine (in Ukrainian).
Ovcharenko, O., Potrokhov, A., Sosnovska, D., Hoysyuk, Yu., Yaroshko, O., Shevchenko, T., Budzanivska, I., Rudas, V. & Kuchuk, M. (2023). Increased virus resistance in transgenic petunia with heterologous ZRNase II gene. JJBS, 16, No. 4, pp. 587-592 https://doi.org/10.54319/jjbs/160403
Slominski, A. T., Li, W., Kim, T.-K., Semak, I., Wang, J., Zjawiony, J. K. & Tuckey, R. C. (2015). Novel activities of CYP11A1 and their potential physiological significance. J. Steroid Biochem. Mol. Biol., 151, pp. 25-37. https://doi.org/10.1016/j.jsbmb.2014.11.010
Sakhno, L. O., Ostapchuk, A. M., Klochko, V. V. & Kuchuk, M. V. (2011). Fatty acid oil composition of canola plants expressing mammalian cytochrome P450SCC cyp11A1 gene. In: Advances in research and technology of rapeseed oil. Monograph–part III (pp. 55-59). Toruń: Wydawnictwo Naukowe Uniwersytetu Mikołaja Kopernika.
Sakhno, L. O. (2011). Seed germination features of canola plants expressing mammalian cytochrome P450SCC cyp11A1 gene. The Bulletin of Vavilov Society of Geneticists and Breeders of Ukraine. 9, No. 2, pp. 253-259 (in Russian).
Sakhno, L. O. (2013). Plant biomass increase: recent advances in genetic engineering. Вiopolym. Cell., 29, No. 6, pp. 443-453. https://doi.org/10.7124/bc.000838
Vasekina, A. V., Yershov, P. V., Reshetova, O. S., Tikhonova, T. V., Lunin, V. G. & Babakov, A. V. (2005). Vacuolar Na+/H+ antiporter from barley: Identification and response to salt stress. Biochemistry, 70, pp. 100-107. https:// doi.org/10.1007/s10541-005-0057-8
Roslyakova, T. V., Lasareva, E. M., Kononenko, N. V., Babakov, A. V. (2009). New isoform HvNHX3 of vacuolar Na+/H+ antiporter in barley: expression and immunolocalization. Biochemistry, 74, No. 5, pp. 549-556. https:// doi.org/10.1134/s0006297909050101
Jabeen, Z., Irshad, F., Hussain, N., Han, Y. & Zhang, G. (2022). NHX-type Na+/H+ antiporter gene expression
under different salt levels and allelic diversity of HvNHX in wild and cultivated barleys. Front. Genet., 12, 809988. https://doi.org/10.3389/fgene.2021.809988
Ovcharenko, O. O., Rudas, V. A., Shcherbak, N. L. & Kuchuk, M. V. (2018). Obtaining of transgenic potato plants (Solanum tuberosum L.) that contain antisense sequence of prolindehydrogenase gene. Faktori eksperimental'noï evolûcìï organìzmìv, 22, pp. 299-304 (in Ukrainian). https://doi.org/10.7124/FEEO.V22.965
Ye, Z-H. & Droste, D. L. (1996). Isolation and characterization of cDNAs encoding xylogenesis-associated and wounding-induced ribonucleases in Zinnia elegans. Plant Mol. Biol., 30, pp. 697-709. https://doi.org/10.1007/ BF00019005
Kyrpa-Nesmiian, T., Rudas, V., Ovcharenko, O., Osipenko, V., Kharhota, M. & Kuchuk, M. (2018). Studies on the adaptation of Solanum tuberosum plants expressing the desA gene to osmotic stress. Proceeding of the 2nd International Conference „Smart Bio“ (p. 72), Kaunas, Lithuania. Retrieved from http://icsb.vdu.lt/wp-content/ uploads/2018/05/ABSTRACT-BOOK-ICSB-2018.pdf
Law of Ukraine “On state regulation of genetic engineering activities and state control over the placing on the market of genetically modified organisms and products”. Vidomosti Verkhovnoyi Rady (VVR), 2023 (in Ukrainian). Retrieved from https://zakon.rada.gov.ua/laws/show/3339-20#Text
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Reports of the National Academy of Sciences of Ukraine
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
