Fiziol. rast. genet. 2023, vol. 55, no. 3, 209-224, doi:

Agrobacterium-mediated transformation of promising winter wheat genotypes in culture in vitro

Dubrovna O.V., Slivka L.V., Velikozhon L.H., Kulesh S.S.

  • Institute of Plant Physiology and Genetics, National Academy of Sciences of Ukraine 31/17 Vasylkivska St., Kyiv, 03022, Ukraine

Plastid terminal oxidase is one of the iron-containing enzymes, electron carriers in the electron transport chain of chloroplasts, the functions of which remain not fully understood even to this day. The presented review examines the structure and specific details of the functioning of plastid terminal oxidase (PTOX) under normal physiological conditions and under the influence of various abiotic stresses. One of the known functions of PTOX is participation in the synthesis of carotenoids. In non-photosynthetic tissues or at early stages of plant development, when photosynthetic electron transport is not fully active, PTOX is the main cofactor for phytoene desaturase and z-carotene desaturase, which participate in the carotenoid desaturation reaction. PTOX also participates in the chlororespiratory mechanism in green plant tissues under stress. In wild-type plants and various mutant forms, the participation of PTOX in counteracting light, temperature, salt stress and their combinations is considered. It is shown that very high expression of the PTOX gene in mutant plants does not always lead to the expected increase in resistance. In contrast to this, a number of data from other authors are given, which showed an increase in resistance in various plants species due to an increase in electron transport through PTOX under the stress impact. This contributed to the reduction of the reactive oxygen species production, the destruction of the D1 protein, and, accordingly, to the preservation of the activity of photosystem II (PS II). The data obtained by the authors on the increased content of PTOX in control plants of high resistance winter wheat varieties are also given. Under the influence of drought, the content of PTOX in these varieties increased even more, and the quantum yield of PS II remained at a higher level. PTOX is thought to function as a stress-triggered safety valve that maintains the oxidation of the PS II acceptor side, thereby helping to protect PS II from photodamage. Thus, PTOX can be used as one of the potential candidates for genetic engineering to increase the stress resistance of agricultural plants.

Keywords: photosynthesis, photorespiration, plastid terminal oxidase, abiotic stress

Fiziol. rast. genet.
2023, vol. 55, no. 3, 209-224

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1. Shewry, P.R. (2009). Wheat. Journal of Experimental Botany, 60, No. 6, pp. 1537-1553.

2. Shiferaw, B., Smale, M., Braun, H., Duveiller, E., Reynolds, M. & Muricho, G. (2013). Crops that feed the world 10. Past successes and future challenges to the role played by wheat in global food security. Food Security, 5, pp. 291-317.

3. Kapoor, D., Bhardwaj, S., Landi, M., Sharma, A., Ramakrishnan, M. & Sharma, A. (2020). The impact of drought in plant metabolism: how to exploit tolerance mechanisms to increase crop production. Applied Science, 10, No. 16, pp. 5692.

4. Kiriziy, D.A. & Stasik, O.O. (2022). Effects of drought and high temperature on physiological and biochemical processes, and productivity of plants. Fyzyolohyia rastenyi i henetyka, 54, No. 2, pp. 95-122 [in Ukrainian].

5. Cattivelli, L., Rizza, F., Badeck, F.-W., Mazzucotelli, E., Mastrangelo, A.M., Francia, E., Mare, C., Tondelli, A. & Stanca, A.M. (2008). Drought tolerance improvement in crop plants: An integrated view from breeding to genomics. Field Crop Research, 105, No. 1, pp. 1-14.

6. Khan, S., Anwar, S., Yu, S., Sun, M., Yang, Z. & Gao, Z. (2019). Development of drought-tolerant transgenic wheat: achievements and limitations. International Journal of Molecular Sciences, 20, p. 3350.

7. Bapela, T., Shimelis, H., Tsilo, T.J. & Mathew, I. (2022). Genetic improvement of wheat for drought tolerance: progress, challenges and opportunities. Plants, 11, p. 1331.

8. El-Mouhamady, A., El-Hawary, M. & Habouh, M. (2023). Transgenic wheat for drought stress tolerance: a review. Middle East Journal of Agriculture Research, 12, No. 1, pp. 77-94.

9. Wang, K., Liu, H., Du, L. & Ye, X. (2017). Generation of marker-free transgenic hexaploid wheat via an Agrobacterium-mediated co-transformation strategy in commercial Chinese wheat varieties. Plant Biotechnology Journal, 15, pp. 614-623.

10. Joshi, R., Anwar, K., Das, P., Singla-Pareek, S. & Pareek, A. (2017). Overview of methods for assessing salinity and drought tolerance of transgenic wheat lines. Wheat Biotechnology. Springer: New York.

11. Anwar, A., Wang, K. & Wang, J. (2021). Expression of Arabidopsis ornithine aminotransferase (AtOAT) encoded gene enhances multiple abiotic stress tolerances in wheat. Plant Cell Reports, 40, No. 7, pp. 1155-1170.

12. Dubrovna, O.V., Priadkina, G.O., Mykhalska, S.I. & Komisarenko, A.G. (2021). Water deficiency tolerance of genetically modified common wheat cv. Zymoyarka, containing a heterologous ornithine-d-aminotransferase gene. Agricultural Science and Practice, 8, No. 1, pp. 25-39.

13. Mykhalska, S.I., Komisarenko, A.G. & Kurchii, V.M. (2021). Genes of proline metabolism in biotechnology of increasing wheat osmostability. Faktory eksperymentalnoy evolyutsiyi orhanizmiv, 28, pp. 94-99 [in Ukrainian].

14. Hossain, M.A., Hoque, M.A., Burritt, D.J. & Fujita, M. (2014). Proline protects plants against abiotic oxidative stress: biochemical and molecular mechanisms. In Ahmad, P. (Ed.). Oxidative Damage to Plants. Antioxidant Networks and Signaling; Academic Press is an imprint of Elsevier, pp. 477-521.

15. Kolupaev, Yu.E., Vainer, A.A. & Yastreb, T.O. (2014). Proline: physiological functions and regulation of its content in plants under stress conditions. Visnyk Kharkivskoho natsionalnoho ahrarnoho universytetu. Seriia Biolohiia, 2, No. 32, pp. 6-22 [in Russian].

16. Meena, М., Divyanshu, K., Kumar, S., Swapnil, P., Andleeb, Z., Vaishali, S., Mukesh, Y. & Upadhyay, R. (2019). Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon, 5, No. 12, p. 02952.

17. Mansour, M.M.F. & Ali, E.F. (2017). Evaluation of proline functions in saline conditions. Phytochemistry, 140, pp. 52-68.

18. Tishchenko, E.N. (2013). Genetic engineering with use of L-proline metabolism genes for increase the of plants osmotolerance. Fiziol. rast. genet., 45, No. 6, pp. 488-500 [in Ukrainian].

19. Dubrovna, O.V., Mikhalska, S.I. & Komisarenko, A.G. (2022). Using of proline metabolism genes in plant genetic engineering. Cytology and Genetics, 56, No. 4, pp. 361-378.

20. Bharathi, J., Anandan, R., Benjamin, L., Muneer, S. & Prakash, M. (2023). Recent trends and advances of RNA interference (RNAi) to improve agricultural crops and enhance their resilience to biotic and abiotic stresses. Plant Physiology and Biochemistry, 194, pp. 600-618.

21. Bilir, O., Gol, D., Hong, Y., McDowell, J.M. & Tor, M. (2022). Small RNA-based plant protection against diseases. Frontiers in Plant Sciense, 13, p. 951097.

22. Halder, K., Chaudhuri, A., Abdin, M.Z. & Datta, A. (2023). Tweaking the small non-coding RNAs to improve desirable traits in plant. International Journal of Molecular Sciences, 24, No. 4, p. 3143.

23. Hernandez-Soto, A. & Chacon-Cerdas, R. (2021). RNAi crop protection advances. International Journal of Molecular Sciences, 22, No. 22, p. 12148.

24. Kaur, R., Choudhury, A., Chauhan, S., Ghosh, A., Tiwari, R. & Rajam, M. (2021). RNA interference and crop protection against biotic stresses. Physiology and Molecular Biology of Plants, 27, No. 10, pp. 2357-2377.

25. Rajam, M.V. (2020). RNA silencing technology: A boon for crop improvement. Journal of Bioscienses, 45, pp. 118-122.

26. Tateishi, Y., Nakagama, T. & Esaka, M. (2005). Osmotolerance and growth stimulation of transgenic tobacco cells accumulating free proline by dehydrogenase expression with double-stranded RNA interference technique. Physiologia Plantarum, 125, pp. 1399-3054.

27. 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 the LBA4404 strain harboring binary vector pBi2E with dsRNA-suppressor of proline dehydrogenase gene. Cytology and Genetics, 48, No. 4, pp. 218-226.

28. Mykhalska, S.I., Sergeeva, L.E., Matveeva, A.Y., Kobernik, N.I., Kochetov, A.V., Tishchenko, E.N. & Morgun, V.V. (2014). The elevation of free proline content in osmotolerant transgenic corn plants with dsRNA suppressor of proline dehydrogenase gene. Fiziol. rast. genet., 46, No. 6, pp. 482-489 [in Russian]. 123456789/159462

29. Komisarenko, A.G., Mykhalska, S.I., Kurchii, V.M. & Tishchenko, O.M. (2016). The characterization transgenic sunflower (Helianthus annuus L.) plants with suppressor of proline dehydrogenase gene. Faktory eksperymentalnoy evolyutsiyi orhanizmiv, 19, pp. 143-147 [in Ukrainian]. view/653

30. Sparks, C., Doherty, A. & Jones, H. (2014). Genetic transformation of wheat via Agrobacterium-mediated DNA delivery. Methods in Molecular Biology, 1099, pp. 235-250.

31. Dubrovna, O.V. & Morgun, B.V. (2018). Current status of research of Agrobacterium-mediated transformation of wheat. Fiziol. rast. genet., 50, No. 3, pp. 187-217 [in Ukrainian].

32. Mamrutha, H.M., Kumar, R., Venkatesh, K., Sharma, P., Kumar, R., Tiwari, V. & Sharma, I. (2014). Genetic transformation of wheat - present status and future potential. Journal Wheat Research, 6, No. 2, pр. 107-119. index.php/JWRReview

33. Kumlehn, J. & Hensel, G. (2009). Genetic transformation technology in the Triticeae. Breeding Science, 59, pp. 553-560.

34. Bavol, A.V., Dubrovna, O.V. & Lyalko, I.I. (2007). Regeneration of plants from the explants of the top of wheat seedlings shoots. Visnyk Ukrayinskoho tovarystva henetykiv i selektsioneriv, 5, No. 1-2, pp. 3-10 [in Ukrainian].

35. Sidorov, V. & Duncan, D. (2009). Agrobacterium-mediated maize transformation: immature embryos versus callus. Methods in Molecular Biology, 526, pp. 47-58.

36. Box, M., Coustham, V., Dean, C. & Mylne, J. (2011). Protocol: A simple phenol-based method for 96-well extraction of high quality RNA from Arabidopsis. Plant Methods, 7, pp. 1-10. content/7/1/7

37. Dubrovna, O.V. & Slivka, L.V. (2020). Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro. Faktory eksperymentalnoy evolyutsiyi orhanizmiv, 26, pp. 190-195 [in Ukrainian].

38. Bates, L.S., Waldren, R.P. & Teare, I.D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soils, 39, pp. 205-207.

39. Ahmad, A., Zhong, H., Wang, W. & Sticklen, M. (2002). Shoot apical meristem: in vitro regeneration and morphogenesis in wheat (Triticum aestivum L.). In vitro Cellular & Development Biological Plant, 38, No. 2, pp. 163-167.

40. Pat. 111284 UA. A method of increasing the regenerative capacity of callus cultures of bread wheat by Agrobacterium-mediated transformation, Dubrovna, O.V., Bavol, A.V., Goncharuk, O.M., Voronova, S.S., Publ. 10.11.2016 [in Ukrainian].