Fiziol. rast. genet. 2024, vol. 56, no. 6, 495-514, doi: https://doi.org/10.15407/frg2024.06.495

Effect of drought on the leaf photoassimilation activity and productivity of genetically modified wheat plants with partial suppression of the proline dehydrogenase gene

Kiriziy D.A., Stasik O.O., Dubrov­na O.V., Sokolovska-Sergiienko O.G., Holoboroda A.S.

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

Under the conditions of a pot experiment, the effects of drought on water deficit, chlorophyll content, chloroplast antioxidant enzymes activity, CO2 assimilation, and stomatal conductance in flag leaves, as well as the grain yield of transgenic plants of the winter bread wheat line UK 322/17 (T3), containing a double-stranded RNA suppressor of the proline dehydrogenase gene, were studied comparring with the original genotype. Drought was imposed on plants of both genotypes by reducing soil moisture to a level of 30 % FC, which was maintained for seven days during the earing—flowering period, and then returned to the control level. Soil moisture for the control plants was maintained at an optimal level of 60—70 % FC. Physiological and biochemical indices were measured on the first day of soil moisture reduction to 30 % FC, on the seventh day at this moisture level, and one week after the restoration of the optimal moisture level. It was found that the drought-treated transgenic plants exhibited better responses to the stressor and its after-effect across all the studied indices compared to the plants of the original genotype. Specifically, the water deficit and chlorophyll content in the leaves of the transgenic plants during drought hardly changed. The net CO2 assimilation rate in the flag leaves of the treated transgenic plants both under drought conditions and during the recovery period was significantly higher than in the treated plants of original line. It was found that with the same stomatal conductivity, the photosynthetic rate in the leaves of the transformants was higher than in the original line plants. Accordingly, the former had an advantage over the latter in the water use efficiency at photosynthesis both under drought conditions and during the period after optimal soil moisture restoration. Although the levels of chloroplast superoxide dismutase and ascorbate peroxidase activities were similar in both genotypes under stress conditions, during the restoration period, the activities in the transgenic plants returned to control levels faster than in the original line plants. A seven-day drought during the critical for wheat earing-flowering period negatively affected the plant productivity of both genotypes, but due to the above-mentioned advantages of the photosynthetic apparatus in the transgenic plants, the degree of this effect for them was significantly less than in the original line. The weight of grain per plant in the transgenic plants was almost one-third higher than in the original line.

Keywords: Triticum aestivum L., drought, transgenic plants, proline, photosynthesis, productivity

Fiziol. rast. genet.
2024, vol. 56, no. 6, 495-514

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References

1. Farooq, M., Wahid, A., Zahra, N., Hafeez, M.B. & Siddique, K.H.M. (2024). Recent advances in plant drought tolerance. J. Plant Growth Regul., 43, pp. 3337-3369. https://doi.org/10.1007/s00344-024-11351-6

2. Neupane, D., Adhikari, P., Bhattarai, D., Rana, B., Ahmed, Z., Sharma, U. & Adhikari, D. (2022). Does climate change affect the yield of the top three cereals and food security in the world? Earth, 3, pp. 45-71. https://doi.org/10.3390/earth3010004

3. Bapela, T., Shimelis, H., Tsilo, T. & Mathew, I. (2022). Genetic improvement of wheat for drought tolerance: Progress, challenges and opportunities. Plants, 11, 1331. https://doi.org/10.3390/plants11101331

4. Kiriziy, D., Kedruk, A. & Stasik, O. (2024). Effects of drought, high temperature and their combinations on the photosynthetic apparatus and plant productivity. In Reg. Adapt. Res. Plants, pp. 1-33. New York: Nova Sci. Publishers. https://doi.org/10.52305/ TXQB2084

5. Pequeno, D.N.L., Hernandez-Ochoa, I.M., Reynolds, M., Sonder, K., MoleroMilan, A., Robertson, R.D., Lopes, M.S., Xiong, W., Kropff, M. & Asseng, S. (2021). Climate impact and adaptation to heat and drought stress of regional and global wheat production. Environ. Res. Lett., 16, 054070. https://doi.org/10.1088/1748-9326/abd970

6. Senapati, N., Stratonovitch, P., Paul, M.J. & Semenov, M.A. (2019). Drought tolerance during reproductive development is important for increasing wheat yield potential under climate change in Europe. J. Exp. Bot., 70, No. 9, pp. 2549-2560. https://doi.org/10.1093/jxb/ery226

7. Sadras, V.O. & Richards, R.A. (2014). Improvement of crop yield in dry environments: benchmarks, levels of organization and the role of nitrogen. J. Exp. Bot., 65, No. 8, pp. 1981-1995. https://doi.org/10.1093/jxb/eru061

8. Wasaya, A., Manzoor, S., Yasir, T.A., Sarwar, N., Mubeen, K., Ismail, I.A., Raza, A., Rehman, A., Hossain, A. & EL Sabagh, A. (2021). Evaluation of fourteen bread wheat (Triticum aestivum L.) genotypes by observing gas exchange parameters, relative water and chlorophyll content, and yield attributes under drought stress. Sustainability, 13, No. 9, 4799. https://doi.org/10.3390/su13094799

9. Morgun, V.V., Dubrovna, O.V. & Morgun, B.V. (2016). The modern biotechnologies of producing wheat plants resistant to stresses. Fiziol. rast. genet., 48, No. 3, pp. 196-213 [in Ukrainian]. https://doi.org/10.15407/frg2016.03.196

10. El-Mouhamady, A., El-Hawary, M. & Habouh, M. (2023). Transgenic wheat for drought stress tolerance: a review. Middle East J. Agricult. Res., 12, No. 1, pp. 77-94. https://doi.org/10.36632/mejar/2023.12.1.7

11. Khan, S., Anwar, S., Yu, S., Sun, M., Yang, Z. & Gao, Z. (2019). Development of drought-tolerant transgenic wheat: achievements and limitations. Int. J. Mol. Sci., 20, 3350. https://doi.org/10.3390/ijms20133350

12. Adel, S. & Carels, N. (2023). Plant tolerance to drought stress with emphasis on wheat. Plants, 12, 2170. https://doi.org/10.3390/plants12112170

13. Alsamman, A.M., Bousba, R., Baum, M., Hamwieh, A. & Fouad, N. (2021). Comprehensive analysis of the gene expression profile of wheat at the crossroads of heat, drought and combined stress. Highlight. BioSci., 4. https://doi.org/10.36462/H.BioSci.202104

14. Jogawat, A. (2019). Osmolytes and their role in abiotic stress tolerance in plants. In Mol. Plant Abiotic Stress: Biol. Biotechnol., pp. 91-104. https://doi.org/10.1002/9781119463665.ch5

15. 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. (pp. 477-521), Antioxidant Networks and Signaling; Academic Press is an Imprint of Elsevier. https://doi.org/10.1016/B978-0-12-799963-0.00016-2

16. Kolupaev, Yu.E., Vainer, A.A. & Yastreb, T.O. (2014). Proline: physiological functions and regulation of the content in plants under stress conditions. Visn. Hark. nac. agrar. univ., Ser. Biol., Iss. 2, pp. 6-22.

17. 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, 02952. https://doi.org/10.1016/j.heliyon.2019.e02952

18. Ghosh, U.K., Islam, M.N., Siddiqui, M.N., Cao, X. & Khan, M.A.R. (2022). Proline, a multifaceted signalling molecule in plant responses to abiotic stress: Understanding the physiological mechanisms. Plant Biol., 24 (2), pp. 227-239. https://doi.org/10.1111/plb.13363

19. Dubrovna, O.V., Mikhalska, S.I. & Komisarenko, A.G. (2022). Using of proline metabolism genes in plant genetic engineering. Cytol. Genet., 56 (4), pp. 361-378. https://doi.org/10.3103/S009545272204003X

20. Dubrovna, O.V., Priadkina, G.O., Mykhalska, S.I. & Komisarenko, A.G. (2022). Drought-tolerance of transgenic winter wheat with partial suppression of the proline dehydrogenase gene. Reg. Mechan. Bio., 13 (4), pp. 385-392. https://doi.org/10.15421/022251

21. Dubrovna, O.V., Stasik, O.O., Priadkina, G.O., Zborivska, O.V. & Sokolovska-Sergienko, O.G. (2020). Resistance of genetically modified wheat plants, containing a double-stranded RNA suppressor of the proline dehydrogenase gene, to soil moisture deficiency. Agricult. Sci. Pract., 7, No. 2, pp. 24-34. https://doi.org/10.15407/agrisp7.02.024

22. Slyvka, L.V. & Dubrovna, O.V. (2021). Genetic transformation of promising genotypes of winter soft wheat by the in planta method. Faktory Еksp. Evol. Оrhanizm., 28, pp. 106-111 [in Ukrainian]. https://doi.org/10.7124/FEEO.v28.1384

23. Dubrovna, O.V. & Priadkina, G.O. (2023). The effect of free proline accumulation on the content of photosynthetic pigments in transgenic wheat plants. Faktory Еksp. Evol. Оrhanizm., 33. pp. 118-122 [in Ukrainian]. https://doi.org/10.7124/FEEO.v33.1578

24. Busch, F.A., Ainsworth, E.A., Amtmann, A., Cavanagh, A.P., Driever, S.M., Ferguson, J.N., Kromdijk, J., Lawson, T., Leakey, A.D.B, Matthews, J.S.A., Meacham-Hensold, K., Vath, R.L., Vialet-Chabrand, S., Walker, B.J. & Papanatsiou, M. (2024). A guide to photosynthetic gas exchange measurements: Fundamental principles, best practice and potential pitfalls. Plant, Cell & Env., 47 (9), pp. 3344-3364. https://doi.org/10.1111/pce.14815

25. Shmatko, I.G., Grigoryuk, I.A., Shvedova, O.E. & Petrenko, N.I. (1985). Determination of the physiological reaction of cereals to deterioration of water availability and temperature increase. IPPG, Kiev [in Russian].

26. Wellburn, A.R. (1994). The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. Plant Physiol., 144, pp. 307-313. https://doi.org/10.1016/S0176-1617(11)81192-2

27. Giannopolitis, C.N. & Ries, S.K. (1977). Superoxide dismutase. Occurrence in higher plants. Plant Physiol., 59, No. 2, pp. 309-314. https://doi.org/10.1104/pp.59.2.309

28. Chen, G.-X. & Asada, K. (1989). Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their and molecular properties. Plant Cell Physiol., 30, No. 7, pp. 987-998. https://doi.org/10.1093/oxfordjournals.pcp.a077844

29. Kolupaev, Y.E., Yastreb, T.O., Ryabchun, N.I., Kokorev, A. I., Kolomatska, V.P. & Dmitriev, A.P. (2023). Redox homeostasis of cereals during acclimation to drought. Theor. Exp. Plant Physiol., 35, pp. 133-168. https://doi.org/10.1007/s40626-023-00271-7

30. Liang, X., Zhang, L., Natarajan, S.K. & Becker, D.F. (2013). Proline mechanisms of stress survival. Antioxid. Redox Signal., 19, pp. 998-1011. https://doi.org/10.1089/ars.2012.5074

31. Qamar, A., Mysore, K. & Senthil-Kumar, M. (2015). Role of proline and pyrroline-5-carboxylate metabolism in plant defense against invading pathogens. Front. Plant Sci., 6, 503. https://doi.org/10.3389/fpls.2015.00503

32. Kolupaev, Yu.E., Karpets, Yu.V. & Kabashnikova, L.F. (2019). Antioxidative system of plants: Cellular compartmentation, protective and signaling functions, mechanisms of regulation. Appl. Biochem. Microbiol., 55 (5), pp. 441-459. https://doi.org/10.1134/S0003683819050089

33. Arora, D., Jain, P., Singh, N., Kaur, H. & Bhatla, S.C. (2016). Mechanisms of nitric oxide crosstalk with reactive oxygen species scavenging enzymes during abiotic stress tolerance in plants. Free Radical Res., 50 (3), pp. 291-303. https://doi.org/10.3109/10715762.2015.1118473

34. Kiriziy, D.A. & Sheheda, I.M. (2019). Nitrogen distribution in the source-sink system of plants and its role in the production process. Plant Physiology and Genetics, 51, No. 2, pp. 114-132 [in Ukrainian]. https://doi.org/10.15407/frg2019.02.114

35. Stasik, O.O., Kiriziy, D.A., Sokolovska-Sergiienko, O.G. & Bondarenko, O.Yu. (2020). Influence of drought on the photosynthetic apparatus activity, senescence rate, and productivity in wheat plants. Plant Physiol. Genet., 52, No. 5, pp. 371-387. https://doi.org/10.15407/frg2020.05.371

36. Kiriziy, D.A., Kedruk, A.S., Sokolovska-Sergiienko, O.G., Dubrovna, O.V. & Stasik, O.O. (2021). Responses of photosynthetic apparatus of genetically modified wheat plants containing a double-stranded RNA suppressor of the proline dehydrogenase gene to drought and high temperature. Plant Physiol. Genet., 53, No. 6, pp. 532-549. https://doi.org/10.15407/frg2021.06.532

37. Pornsiriwong, W., Estavillo, G.M., Chan, K.X., Tee, E.E., Ganguly, D., Crisp, P.A., Phua, S.Y., Zhao, C., Qiu, J., Park, J., Yong, M.T., Nisar, N., Yadav, A.K., Schwessinger, B., Rathjen, J., Cazzonelli, C.I., Wilson, P.B., Gilliham, M., Chen, Z.-H. & Pogson, B.J. (2017). A chloroplast retrograde signal, 3¢-phosphoadenosine-5¢-phosphate, acts as a secondary messenger in abscisic acid signaling in stomatal closure and germination. eLife, 6, e23361. https://doi.org/10.7554/eLife.23361

38. Lawson, T. & Matthews, J. (2020). Guard cell metabolism and stomatal function. Annu. Rev. Plant Biol., 71, pp. 273-302. https://doi.org/10.1146/annurev-arplant-050718-100251

39. Condon, A.G. (2020). Drying times: plant traits to improve crop water use efficiency and yield. J. Exp. Bot., 71, No. 7, pp. 2239-2252. https://doi.org/10.1093/jxb/eraa002

40. Bertolino, L.T., Caine, R.S. & Gray, J.E. (2019). Impact of stomatal density and morphology on water-use efficiency in a changing world. Front. Plant Sci., 10, 225. https://doi.org/10.3389/fpls.2019.00225

41. Isgandarova, T.Y., Rustamova, S.M., Aliyeva, D.R., Rzayev, F.H., Gasimov, E.K. & Huseynova, I.M. (2024). Antioxidant and ultrastructural alterations in wheat during drought-induced leaf senescence. Agronomy, 14, 2924. https://doi.org/10.3390/agronomy14122924

42. Krieger-Liszkay, A., Krupinska, K. & Shimakawa, G. (2019). The impact of photosynthesis on initiation of leaf senescence. Physiol. Plant., 166, pp. 148-164. https://doi.org/10.1111/ppl.12921

43. Lopez, M.A., Xavier, A. & Rainey, K.M. (2019). Phenotypic variation and genetic architecture for photosynthesis and water use efficiency in soybean (Glycine max L. Merr). Front. Plant Sci., 10. https://doi.org/10.3389/fpls.2019.00680

44. Carvalho, K., Campos, M.K., Domingues, D.S., Pereira, L.F.P. & Vieira, L.G.E. (2013). The accumulation of endogenous proline induces changes in gene expression of several antioxidant enzymes in leaves of transgenic Swingle citrumelo. Mol. Biol. Rep., 40(4), pp. 3269-3279. https://doi.org/10.1007/s11033-012-2402-5

45. Szekely, G., Abraham, E., Cseplo, A., Rigo, G., Zsigmond, L., Csiszar, J., Ayaydin, F., Strizhov, N., Jasik, J., Schmelzer, E., Koncz, C. & Szabados, L. (2008). Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J., 53 (1), pp. 11-28. https://doi.org/10.1111/j.1365-313X.2007.03318.x

46. Kolupaev, Y.E., Karpets, Y.V., Yastreb, T.O., Shemet, S.A. & Bhardwaj, R. (2020). Antioxidant system and plant cross-adaptation against metal excess and other environmental stressors. In: Landi, M., Shemet, S.A., Fedenko, V.S., (Еds.). Metal toxicity in higher plants. (pp. 21-66). New York: Nova Science Publishers.