Biopolym. Cell. 2020; 36(6):423-432.
Molecular and Cell Biotechnologies
Creation of winter rapeseed Brassica napus L. commercial line of biotechnological plants, resistant to the glyphosate action
1, 2Hnatiuk I. S., 1, 2Varchenko O. I., 1Kuchuk M. V., 2, 3Parii M. F., 1, 2Symonenko Yu. V.
  1. Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
    148, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03143
  2. Ltd “Ukrainian Scientific Institute of Plant Breeding” (VNIS)
    30, Vasylkivska Str., Kyiv, Ukraine, 03022
  3. National University of Life and Environmental Sciences of Ukraine
    15, Heroiv Oborony Str., Kyiv, Ukraine, 03041

Abstract

Aim. To create biotechnological plants of a commercial line winter rapeseed, which carry the glyphosate and phosphinostricin resistance genes. Methods. In vitro tissue culture method, Agrobacterium-mediated genetic transformation, polymerase chain reaction and χ2 methods were used. Results. The PCR analysis results are presented for the biotechnological winter rapeseed plants obtained after genetic transformation to confirm the presence of CP4 epsps gene insert in 11 of 12 plants, as well as the presence of bar gene integration in 6 of 7 plants. All obtained regenerant plants were adapted to in vivo conditions, treated with glyphosate and vernalized. The obtained seeds were sterile germinated and the disjoining was determined based on the resistance to the selective agent. Conclusion. 11 biotechnological plants with the glyphosate (epsps) and kanamycin (nptII) resistance genes were obtained, as well as 6 biotechnological plants, which carry [the] phosphinothricin (bar) resistance genes in addition to the epsps gene. The presence of the CP4 EPSPS protein in transgenic rapeseed plants has been proven by the molecular analysis of protein expression using an immunoassay system. The expression of selective agents resistance genes was confirmed in T1 generation.
Keywords: Brassica napus, winter rapeseed, bar gene, epsps gene, nptII gene, genetic transformation

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