Russian Federation
Russian Federation
Russian Federation
Russian Federation
Russian Federation
Russian Federation
UDC 57.043
Modern breeding of cereals such as wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) is complemented by biotechnological methods, including in vitro cell selection and genetic transformation. For long-term preservation of the resulting unique cell lines and minimisation of genetic changes, the scientists should use cryopreservation, which halts metabolism and eliminates the need for regular passages. The aim of this study was to evaluate the efficiency of cryopreservation media (glycerol, glycerol and apple pectin) for preserving spring barley and wheat callus cells at –80 °C in a domestic electric freezer. Barley (four lines) and wheat (two lines) calli obtained from immature embryos were used. Callus cells were frozen under the protection of cryopreservative media in electric freezer at –80 °C for seven days, followed by cultivation of thawed callus cells in a nutrient medium. Before cooling and after thawing, we assessed the viability of callus cells using a vital dye. The ability of cells to regenerate after thawing was also assessed. After exposure to subzero temperatures using cryopreservation solutions, the cell membrane integrity of both barley and wheat calli, regardless of genotype, was consistently maintained at 50 %. Thus, the use of cryopreservation media (glycerol, glycerol and apple pectin) for preserving spring wheat and barley callus cells at –80 °C in a domestic electric freezer is successful in maintaining the cell membrane integrity of callus cells.
wheat callus, barley callus, apple pectin, glycerol, cell membrane integrity, genotype, cultivation
1. Özbek, K. Advances in wheat breeding. Chapter: Wheat Genetic Resources / K. Özbek, C. N. Keskin, N. Zencirci. – Singapore: Springer, 2024. – P. 525–554.
2. Shupletsova, O. N. Geneticheskie istochniki selekcii yachmenya Hordeum vulgare L. v Volgo-Vyatskom regione [Genetic sources of barley (Hordeum vulgare L.) breeding in the Volga-Vyatka region] / O. N. Schupletsova, I. N. Shchennikova // Trudi po prikladnoi botanike, genetike i selekcii [Transactions on Applied Botany, Genetics, and Breeding]. – 2019. – Vol.180, № 1. – P. 82–88. DOI: https://doi.org/10.30901/2227-8834-2019-1-82-88; EDN: https://elibrary.ru/SUMQUM
3. Volkova, L. V. Rezultati sravnitelnogo izucheniya kommercheskih sortov yarovoi myagkoi pshenici v usloviyah Kirovskoi oblasti [Results of a comparative study of commercial varieties of spring soft wheat in the Kirov region] / L. V. Volkova //Agrarnaya nauka Evro-Severo-Vostoka [Agricultural Science of the Euro-North-East]. – 2025. – Vol. 26, № 3. – P. 536–545. DOI: https://doi.org/10.30766/2072-9081.2025.26.3.536-545; EDN: https://elibrary.ru/LFQTFQ
4. Freezing of dehydrated calli of spring wheat (Triticum aestivum L.) in liquid nitrogen and their morphogenetic potential / A. I. Solovyeva, O. N. Vysotskaya, A. S. Popov [et al.] // Biology Bulletin. – 2010. – Vol. 37, № 5. – P. 489–495. DOI: https://doi.org/10.1134/S1062359010050080; EDN: https://elibrary.ru/MXQFAN
5. A simple and efficient protocol for cryopreservation of Taxodium hybrid ‘zhongshanshan’ embryogenic callus / T. Chen, X. Jia, C. Yu [et al.] // Plant Cell, Tissue and Organ Culture (PCTOC). – 2024. – Vol.156, № 44. DOI: https://doi.org/10.1007/s11240-023-02667-4
6. Tissue culture-induced heritable genomic variation in rice, and their phenotypic implications / D. Zhang , Z. Wang, N. Wang [et al.] // PLoS ONE. – 2014. – Vol. 9. – P. e96879. DOI: https://doi.org/10.1371/journal.pone.0096879
7. Benson, E. E. Cryopreservation of phytodiversity: A critical appraisal of theory & practice / E. E. Benson // Critical Reviews in Plant Sciences. – 2008. – Vol. 27, № 3. – P. 141–219. DOI: https://doi.org/10.1080/07352680802202034
8. Cryopreservation of shoot apices and callus cultures of globe artichoke using vitrification method / S. A. Bekheet, V. Sota, H. M. El-Shabrawi [et al.] // Journal of Genetic Engineering and Biotechnology. – 2020. – Vol. 18. – № 1. DOI: https://doi.org/10.1186/s43141-019-0016-1
9. Hao, Y.-J. Effects of cryopreservation on developmental competency, cytological and molecular stability of citrus callus / Y.-J. Hao, C.-X. You, X.-X. Deng // Cryo-Letters. – 2002. – Vol. 23, № 1. – P. 27–35.
10. Elliott, G. D. Cryoprotectants: A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures / G. D. Elliott, S. Wang, B. J. Fuller // Cryobiology. – Vol. 76. – P. 74–91. DOI: https://doi.org/10.1016/j.cryobiol.2017.04.004
11. Murashige, T. A. Revised medium for rapid growth and bio assays with tobacco tissue culture / T.A. Murashige, F. Scoog // Physiologia Plantarum. – 1962. – Vol. 15. – P. 473–497. DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
12. Defense-related callose deposition in plants against pathogens: A review / S. R. Roohall, F. Fariba, G. V. Mozhgan [et al.] // International Journal of Biological Macromolecules. – 2025. – Vol. 320, № 4. – P. 146005. DOI: https://doi.org/10.1016/j.ijbiomac.2025.146005
13. Uemura, M. Survival of carnation (Dianthus caryophyllus L.) shoot apices frozen to the temperature of liquid nitrogen / M. Uemura, A. Sakai // Plant and Cell Physiology. – 1980. – Vol. 21. – № 1. – P. 85–94. EDN: https://elibrary.ru/IVRVPR
14. Sakai, A. Cryopreservation of nucellar cells of navel orange (Citrus sinensis var. brasilensis Tanaka) by vitrification / A. Sakai, S. Kobayashi, I. Oiyama // Plant Cell Reports. – 1990. – Vol. 9, № 3. – P. 30–33. DOI: https://doi.org/10.1007/bf00232130; EDN: https://elibrary.ru/MDRBXP
15. Panis, B. Cryopreservation of plant germplasm / B. Panis, R. Swennen, F. Engelmann // Acta Horticulturae. – 2001. – Vol. 560. – P. 79–86. DOI: https://doi.org/10.17660/ActaHortic.2001.560.8
16. Direct observation of common cryoprotectant permeation into rice callus by CARS microscopy / F. M. D. Samuels, K. C. Pearce, S. Soderlund [et al.] // Cell Reports Physical Science. – 2023. – Vol. 4. – № 7. – P. 101469. DOI: https://doi.org/10.1016/j.xcrp.2023.101469; EDN: https://elibrary.ru/RNLJES
17. Apple pectin as a new component for cryopreservation of nucleated cells / M. I. Sergushkina, O. O. Zaitseva, A. N. Khudyakov [et al.] // Biopreservation and Biobanking. – 2021. – Vol. 20, № 1. – P. 84–89. DOI: https://doi.org/10.1089/bio.2021.0004
