Agrobacterium-mediated transformation of spring camelina with yeast genes of trehalose biosynthesis

  • A. Yu. Kvasko Institute of Food Biotechnology and Genomics NAS of Ukraine, Ukraine, 04123, Kyiv, Osypovskogo str., 2A https://orcid.org/0000-0001-5014-3630
  • A. S. Lazarets Educational and Scientific Center “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine, 01601, Kyiv, Volodymyrska str., 64/13
  • S. V. Isayenkov Institute of Food Biotechnology and Genomics NAS of Ukraine, Ukraine, 04123, Kyiv, Osypovskogo str., 2A https://orcid.org/0000-0002-3119-1642
  • A. I. Yemets Institute of Food Biotechnology and Genomics NAS of Ukraine, Ukraine, 04123, Kyiv, Osypovskogo str., 2A https://orcid.org/0000-0001-6887-0705
Keywords: Camelina sativa, trehalose, TPS1, TPS2 gene

Abstract

Aim. The aim of the study was the obtaining of Camelina sativa (L.) Crantz lines with yeast genes of trehalose synthesis TPS1 and TPS2 to increase their resistance to drought. Methods. Seeds of C. sativa genotype FEORZhYaF-1 were used for in vitro culture establishment. For this hypocotyl segments and shoot meristems of 5-days-old camelina seedlings were cultivated on three different nutrient media for regeneration supplemented with various hormone combinations. Vector constructions pGWB2-TPS1 and pGWB2-TPS2 with TPS1 and TPS2 genes have been used for genetic transformation. Results. The highest efficiency of plant regeneration from hypocotyl explants was found on medium supplemented with 1 mg/l BAP and 0.1 mg/l NAA, and from meristem explants – on medium with 1.5 mg/l BAP and 0.5 mg/l NAA. Agrobacterium-mediated transformation was conducted out, and camelina lines were picked up on corresponding medium with selective concentration of hygromycin. Transgenic nature of obtained plants was confirmed by PCR-analysis. Conclusions. The efficiency of in vitro plant regeneration of C. sativa genotype FEORZhYaF-1 has been investigated. Two types of explants and two vector constructions pGWB2-TPS1 and pGWB2-TPS2 with TPS1 and TPS2 yeast trehalose synthesis genes have been used for obtaining of transgenic camelina lines.

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