Physiological and biochemical analysis of transgenic wheat plants of seed generation T2 with heterologous ornithine-δ-aminotransferase gene

  • O. V. Dubrovna
  • L. V. Slivka
  • S. S. Kulesh
DOI: https://doi.org/10.7124/FEEO.v24.1081

Abstract

Aim. To carry out physiological and biochemical analysis of genetically modified plants of bread wheat of seed generation T2 with the heterologous ornithine-Δ-aminotransferase gene. Methods. Biochemical determination of free L-proline content and ornithine-Δ-aminotransferase enzyme activity; physiological examination of plant growth in in vitro and in vivo conditions. Results. It was shown that transgenic plants did not differ from the controls under optimal conditions of cultivation. It was determined that transgenic plants under the conditions of osmotic stress are characterized by faster growth in comparison with control genotypes. It was found that T2 plants differed in the increased activity of the ornithine-Δ-aminotransferase enzyme, which is manifested when the norm-stress-norm conditions change. It was found that the introduction of a genetic construct that increases the expression of the oat gene does not lead to a significant change in the level of free L-proline in the leaves of plants, either in normal, or in terms of osmotic stress. Conclusions. Changes in the metabolism of transgenic plants allow them to better adapt to adverse conditions. They have better adaptive plasticity, since the yield of most of the transformed lines was significantly higher, compared to non-transformed plants that were under groundwater deficit.

Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine-Δ-aminotransferase gene, T2 plants, physiological-biochemical analysis.

References

Goldenkova-Pavlova I.V., Mirahorli N., Maali A.R., Isaenko E., Kartel N.A., Yuryeva N.O., Abdeyeva I.A. Experimental models for the creation of transgenic plants resistant to stress factors. Cytology and Genetics. 2007. Vol. 41 (3). P. 44–49. [in Russian]

Morgun B.V., Tishchenko E.N. Molecular Biotechnologies for Increasing the Sustainability of Cultivated Cereals to Osmotic Stress K.: Logos, 2014. 218 p. [in Russian]

Tishchenko O.M., Mikhals'ka S.I., Morgun B.V. Genetic engineering and cell selection for enhancing osmotolerance of culti-vated plants. Plant physiology and genetics. 2016. Vol. 48 (3). P. 257–266. [in Ukrainian]

Hiei Y., Ishida Y., Komary T. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens. Frontiers in Plant Science. 2014. Vol. 5. P. 628. doi: 10.3389/fpls.2014.00628.

Fleury D., Jefferies S., Kuchel H., Langridge P. Genetic and genomic tools to improve drought tolerance in wheat. J Exp Bot. 2010. Vol. 61 (12). P. 3211–3222. doi: 10.1093/jxb/erq152.

Mamrutha H., Rakesh K., Karnam V., Sharma P., Kumar R., Tiwari V. Genetic transformation of wheat– рresent status and future potential. J. of Wheat Research. 2014. Vol. 6, (2). Р. 107–119.

Binka F., Orczyk W., Nadolska-Orczyk A. The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection Cassettes. J. of Applied Genetics. 2012. Vol. 53. Р. 1–8. doi: 10.1007/s13353-011-0064-y.

Dubrovna O.V., Morgun B.V. The current state of the research of Agrobacterium-mediated wheat transformation. Plant physi-ology and genetics. Vol. 50, No. 3. P. 187–217. [in Ukrainian]

Stranska J., Kopecny D., Tylichova M., Snégaroff J, Šebela M. Ornithine delta-aminotransferase: An enzyme implicated in salt tolerance in higher plants. Plant Signal Behav. 2008. Vol. 3 (11). P. 929–935.

Roosens N.H., Bitar F.A., Loenders K., Angenon G., Jacobs M. Overexpression of ornthine-aminotransferase increases proline biosynthesis and confers osmotolerance in transgenic plants. Mol. Breed. 2002. Vol. 9. P. 73–80.

Wu L., Fan Z., Guo L. Over-expression of an Arabidopsis OAT gene enhances salt and drought tolerance in transgenic rice. Chinese Sci. Bull. 2003. Vol. 48 (23). P. 2594–2600.

Vendruscolo E., Schuster I., Pileggi M. 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.

Goncharuk O.M., Dubrovna O.V. Receipt of Genetically-Modified Wheat Plants with the Ornithine-Δ-Aminotransferase het-erologous gene. Factors of Expediental Evolution of Organisms. 2018. Vol. 22. P. 222–227. [in Ukrainian]

Andriushchenko V.K., Saianova V.V., Zhuchenko A.A., D'iachenko N.I., Chilikina L.A., Drozdov V.V., Korochkina S.K., Cherep G.I., Medvedev V.V., Niutin Iu.I. Modification of the method for determining proline to identify drought-resistant forms of the genus Lycopersicon Tourn. Izvestiia Akademii Nauk Moldavskoi SSR. 1981. Vol. 4. P. 55–60. [in Russian]

Vogel R.H., Kopac M. Some propertiesof ornithine sect-transaminase from Neurospora. Biochim. Biophys. Acta. 1960. Vol. 37. P. 539–540.

Tishchenko E.N. Genetic engineering using the L-proline metabolism genes to enhance osmotic tolerance of plants. Plant Physiology and Genetics. 2013. Vol. 45 (6). P. 488–500. [in Russian]