The effect of free proline accumulation on the content of photosynthetic pigments in transgenic wheat plants

DOI: https://doi.org/10.7124/FEEO.v33.1578
Keywords: Triticum aestivum, Agrobacterium-mediated transformation, proline, chlorophyll

Abstract

Aim. To analyze the effect of L-proline accumulation on chlorophyll content in transgenic wheat plants with a double-stranded RNA suppressor of the proline dehydrogenase gene under physiological and stress conditions. Methods. Agrobacterium-mediated transformation in planta; biochemical determination of free proline content; spectrophotometric determination of photosynthetic pigments content; of mathematical statistics. Results. It was shown that the content of free proline in the leaves of transgenic plants under physiological conditions was 1.7-1.9 times higher compared to the original genotype. Under conditions of drought, the content of this aminoacid increased in non-transformed plants of the original genotypes by 2.9-3.1 times, and in transgenic plants – by 4.5-4.9 times. The amount of chlorophyll in the flag leaves of plants of the original genotypes and their transgenic lines under physiological conditions did not differ significantly, while under drought conditions in the first of them it was 1.1-1.2 times less than in the second. Drought reduced the total chlorophyll content in plants of the original genotype to 85-90 %, compared to physiological conditions, while no significant changes were found in transgenic plants. Conclusions. It was established that under conditions of soil drought, the increase in proline content in genetically modified wheat plants compared to non-transgenic ones is accompanied by an increase in the amount of total chlorophyll (by 10-15 %), which indicates a better efficiency of their pigment apparatus under stressful conditions.

References

Ghosh U. K., Islam M. N., Siddiqui M. N., Cao X., Khan M. A. R. Proline, a multifaceted signalling molecule in plant responses to abiotic stress: understanding the physiological mechanisms. Plant Biology. 2022. Vol. 24 (2). P. 227–239. doi: 10.1111/plb.13363.

Dubrovna O. V., Mykhalska S. I., Komisarenko A. G. Using proline metabolism genes in plant genetic engineering. Cytology and Genetics. 2022. Vol. 56 (4). P. 361–378. doi: 10.3103/S009545272204003X.

Tateishi Y., Nakagama T., Esaka M. Osmotolerance and growth stimulation of transgenic tobacco cells accumulating free proline by dehydrogenase expression with double-stranded RNA interference technique. Physiologia Plantarum. 2005. Vol. 125. P. 1399–3054. doi: 10.1111/j.1399-3054.2005.00553.x.

Komisarenko A. G., Mykhalska S. I., Kurchii V. M., Tishchenko O. M. The characterization transgenic sunflower (Helianthus annuus L.) plants with suppressor of proline dehydrogenase gene. Factors in experimental evolution of organisms. 2016. Vol. 19. P. 143–147. [in Ukrainian]

Mykhalska S. I., Sergeeva L. E., Matveeva A. Yu., Kobernik N. I., Kochetov A. V., Tishchenko E. N., Morgun V. V. The elevation of free proline content in osmotolerant transgenic corn plants with dsRNA suppressor of proline dehydrogenase gene. Plant Physiology and Genetics. 2014. Vol. 46 (6). P. 482–489. Retrieved from: http://dspace.nbuv.gov.ua/handle/123456789/159462. [in Russian]

Rehman S., Bilal M., Rana R., Tahir N., Shah M., Ayalew H., Yan G. Cell membrane stability and chlorophyll content variation in wheat (Triticum aestivum L.) genotypes under heat and drought conditions. Crop and Pasture Science. 2016. Vol. 67. P. 712–718. doi: 10.1071/CP15385.

Chen L., Shenai P., Zheng F., Somoza A., Zhao Y. Optimal Energy Transfer in Light-Harvesting Systems. Molecules. 2015. Vol. 20 (8). P. 15224–15272. doi: 10.3390/molecules200815224.

Othmani A., Ayed S., Slama-Ayed O., Slim-Amara H., Younes M. B. Durum wheat response (Triticum durum Desf.) to drought stress under laboratory conditions. Journal of Agriculture and Veterinary Science. 2019. Vol. 12 (2). P. 3–6. doi: 10.9790/2380-1202010104.

Sharma V., Kumar A., Chaudhary A., Mishra A., Rawat S., Basavaraj Y. B., Shami V., Kaushik P. Response of wheat genotypes to drought stress stimulated by PEG. Stresses. 2022. Vol. 2 (1). P. 26–51. doi: 10.3390/stresses2010003.

Bekka S., Abrous-Belbachir O., Djebbar R. Effects of exogenous proline on the physiological characteristics of Triticum aestivum L. and Lens culinaris Medik. under drought stress. Acta Agriculturale Slovenica. 2018. Vol. 111 (2). P. 477–491. doi: 10.14720/aas.2018.111.2.20.

Chumakov M. I., Moiseeva E. M. Technologies of Agrobacterium plant transformation in planta. Applied Biochemistry and Microbiology. 2012. Vol. 48. P. 657–666. doi: 10.1134/S0003683812080017.

Bates L. S., Waldren R. P., Teare, I. D. Rapid determination of free proline for water-stress studies. Plant and Soils. 1973 Vol. 39. P. 205–207. doi: 10.1007/BF00018060.

Wellburn A. P. The spectral determination of chlorophyll a and b, as well as carotenoids using various solvents with spectrophotometers of different resolution. Journal of Plant Physiology. 1994. Vol. 144 (3). P. 307–313. doi: 10.1016/S0176-1617(11)81192-2.

Noor S., Ali S., Rehman H., Ullah F., Ali, G. M. Comparative study of transgenic (DREB1A) and non-transgenic wheat lines on relative water content, sugar, proline and chlorophyll under drought and salt stresses. Sarhad Journal of Agriculture. 2018. Vol. 34 (4). P. 986–993. doi: 10.17582/journal.sja/2018/34.4.986.993.

Gao H., Wang Y., Xu P., Zhang Z. Overexpression of a WRKY transcription factor TaWRKY2 enhances drought stress tolerance in transgenic wheat. Frontiers in Plant Science. 2018. Vol. 9. P. 997. doi: 10.3389/fpls.2018.00997.

Yu T. T., Xu Z. Z., Guo J. J., Wang Y. Y., Abernathy B., Fu J. J., Chen X., Zhou Y. Y., Chen M., Ye X. X. Improved drought tolerance in wheat plants overexpressing a synthetic bacterial cold shock protein gene SeCspA. Scientific Reports. 2017. Vol. 7. P. 27–25. doi: 10.1038/srep44050.

Meena М., Divyanshu K., Kumar S. Swapnil P., Zehra A., Shukla V., Yadav M., Upadhyay R. S. Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon. 2019. Vol. 5 (12). 02952. doi: 10.1016/j.heliyon.2019.e02952.

Kaur G., Asthir B., Bains N. Modulation of proline metabolism under drought and salt stress conditions in wheat seedlings. Indian Journal of Biochemistry and Biophysics. 2018. Vol. 55. P. 114–124: Retrieved from: http://nopr.niscpr.res.in/handle/123456789/44345.