Physiological, biochemical and economic characteristics of transgenic winter wheat plants with RNA suppressor of the proline dehydrogenase gene
Abstract
Aim. To analyze the physiological, biochemical and economic characteristics of genetically modified plants of new promising genotypes of winter bread wheat of seed generation T2 with a double-stranded RNA suppressor of the proline dehydrogenase gene. Methods. Agrobacterium-mediated transformation in vitro; biochemical determination of proline dehydrogenase enzyme activity and free proline content; morphometric indicators and elements of crop structure; mathematical statistics. Results. It is shown that transgenic plants, in contrast to control, grow on a medium with mannitol more intensely, retaining a green color. It was found that both under normal conditions and under conditions of water deficiency, plants of seed generation T2 have an increased level of free Proline in the leaves compared to control genotypes. It was found that transformants are characterized by reduced activity of the enzyme proline dehydrogenase, which is manifested by changes in normal – stress – normal conditions. Transgenic T2 plants had a higher tolerance to water deficiency compared to the original, which was reflected in the nature of their growth. In conditions of soil moisture deficiency, the yield of most transformed lines was higher compared to untransformed plants. Conclusions. The results suggest that the use of a vector construct with a double-stranded RNA suppressor of the ProDH gene is effective for creating transgenic winter bread wheat plants with increased tolerance to water deficiency.
References
Hiei Y., Ishida Y., Komari T. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens. Frontiers in Plant Sci. 2014. Vol. 5. P. 1–11. doi: 10.3389/fpls.2014.00628.
Vendruscolo E.C., Schuster I., Pileggi M., Scapim C.A., Molinari H.B., Marur C.J., Vieira L.G. Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat. Plant Physiol. 2007. Vol 164 (10). P. 1367–1376. doi: 10.1016/j.jplph.2007.05.001.
Anwar A., She M., Wang K., Riaz B., Ye X. Biological roles of ornithine aminotransferase (OAT) in plant stress tolerance: present progress and future perspectives. Int J Mol Sci. 2018. 19. P. 3681. doi: 10.3390/ijms19113681.
Szabados L., Savoure A. Proline: A multifunctional amino acid. Trends Plant Sci. 2009. Vol. 15. P. 89–97. doi: 10.1016/j.tplants.2009.11.009.
Sharma S., Villamor J.G., Verslues P.E. Essential role of tissue-specific proline synthesis and catabolism in growth and redox balance at low water potential. Plant Physiol. 2011. Vol. 157. P. 292–304. doi: 10.1104/pp.111.183210.
Tishchenko E.N. Genetic engineering using genes of L-proline metabolism to increase the osmotolerance of plants. Plant Physiology and Genetics. 2013. Vol. 45 (6). P. 488–500. Retrieved from: http://dspace.nbuv.gov.ua/handle/123456789/159371. [in Russian]
Dubrovna O.V., Stasik O.O., Priadkina G.O., Zborivska O.V., Sokolovska-Sergienko O.G. Resistance of genetically modified wheat plants, containing a double-stranded RNA suppressor of the proline dehydrogenase gene, to soil moisture deficiency. Agricultural Science and Practice. 2020. Vol. 7 (2). P. 24–34. doi: 10.15407/agrisp7.02.024.
Roosens N.H., Bitar F.A., Loenders K. Overexpression of ornithine-aminotransferase increases proline biosynthesis and confers osmotolerance in transgenic plants. Mol Breed. 2002. Vol. 9 (2). P. 73–80. doi: 10.1023/A %3A1026791932238.
Ibragimova Ya.S., Gerasimova S.V., Kochetov A.V. The role of the proline dehydrogenase gene in maintaining stress resistance in plants. Plant physiology. 2012. Vol. 59 (1). P. 99–107. doi: 10.1104/pp.110.167163. [in Russian]
Servet C., Ghelis T., Richard L. Proline dehydrogenase: a key enzyme in controlling cellular homeostasis. Front Biosci. 2012. Vol. 1 (17). P. 607–620. doi: 10.2741/3947.
Borsani O., Zhu J., Verslues E.P. Endogenous siRNAs derived from a pair of natural cis-antisense transcripts regulate salt tolerance in Arabidopsis. Cell. 2005. Vol. 123. P. 1279–1291. doi: 10.1016/j.cell.2005.11.035.
Manavalan L.P., Chen X., Clarke J. RNAi-mediated disruption squalen synthase improves drought tolerance and yield in rise. J Exp Bot. 2012. Vol. 63. P. 163–175. doi: 10.1093/jxb/err258.
Dubrovna O.V., Slivka L.V. Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro. Factors of experimental evolution of organisms. 2020. Vol. 26. C. 190–195. doi: 10.7124/FEEO.v26.1264. [in Ukrainian]
Bates L.S., Waldren R.P., Teare I.D. Rapid determination of free proline for water-stress studies. Plant Soil. 1973. Vol. 39. P. 205–207. doi: 10.1007/BF00018060.
Mattioni C., Lacerenza N.G., Troccoli A.D., De Leonardis A.M., Di Fonzo N. Water and solt stress-induced alterations in proline metabolism of Triticum durum seedlings. Physiol Plant. 1997. Vol. 101. P. 787–792. doi: 10.1111/j.1399-3054.1997.tb01064.x.
