Genetic transformation of Nicotiana tabacum with the yeast TPS1 and TPS2 genes involved in trehalose synthesis

  • A. Yu. Kvasko Institute of Food Biotechnology and Genomics NAS of Ukraine Ukraine 04123, Kyiv, Osipovskogo st., 123
  • S. V. Isayenkov Institute of Food Biotechnology and Genomics NAS of Ukraine Ukraine 04123, Kyiv, Osipovskogo st., 123
  • E. E. Krasnoperova Institute of Food Biotechnology and Genomics NAS of Ukraine Ukraine 04123, Kyiv, Osipovskogo st., 123
  • K. V. Dmytruk Іnstitute of Cell Biology NAS of Ukraine Ukraine, 79005, Lviv, Dragomanova st., 14/16
  • A. I. Yemets Institute of Food Biotechnology and Genomics NAS of Ukraine, Ukraine, 04123, Kyiv, Osipovskogo st., 123
DOI: https://doi.org/10.7124/visnyk.utgis.17.2.1215

Abstract

Aim. The aim of the study was the creation of vector constructions with yeast genes of trehalose synthesis TPS1 and TPS2 and their using for Agrobacterium-mediated transformation of N. tabacum. Methods. Strain of Agrobacterium tumefaciens GV3101 carrying vector constructions — pGWB2-TPS1 and pGWB2-TPS2 with TPS1 and TPS2 target genes respectively under 35S promoter of cauliflower mosaic virus and selectable hpt gene of hygromycin phosphotransferase has been used for plants transformation. N. tabacum leaf explants were used for Agrobacterium-mediated transformation. The medium with the addition of hygromycin was applied to transgenic lines selection. Results. The created vector constructions pGWB2-TPS1 and pGWB2-TPS2 has been used in genetic N. tabacum transformation. Target genes TPS1 and TPS2 were integrated applied of Agrobacterium-mediated transformation and transgenic lines of N. tabacum were selected with addition at 25 mg/L hygromycin into the medium. Molecular analysis confirmed the transgenic nature of selected lines. Conclusions. Sensitivity of selected lines for sugars content into the medium was established for shoots and roots formation of tobacco plants. Conditions for increase of transformation frequency and rooting of transgenic lines of plant after integrated of target TPS1 and TPS2 genes were investigated.
Keywords: trehalose, yeast genes TPS1, TPS2, genetic transformation, Nicotiana tabacum.

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