Determination of phosphinothricin and paromomycin selective concentrations for obtaining transgenic spelt plants

  • I. O. Nitovska Institute of Cell Biology and Genetic Engineering of Nat. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnoho str., 148 https://orcid.org/0000-0002-8976-5725
  • A. S. Panasenko Institute of Cell Biology and Genetic Engineering of Nat. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnoho str., 148 https://orcid.org/0009-0007-9802-2679
  • V. Yu. Redko National University of Kyiv-Mohyla Academy, Ukraine, 04070, Kyiv, Skovorody, str., 2
  • M. I. Zadorozhna National University of Kyiv-Mohyla Academy, Ukraine, 04070, Kyiv, Skovorody, str., 2 https://orcid.org/0009-0003-4606-0156
  • B. V. Morgun Institute of Cell Biology and Genetic Engineering of Nat. Acad. Sci. of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnoho str., 148 https://orcid.org/0000-0001-7041-6894
DOI: https://doi.org/10.7124/FEEO.v34.1640
Keywords: Triticum spelta L., spelt wheat, shoot apical meristem culture, mature embryo culture, plant genetic transformation

Abstract

Aim. To determine the selective concentrations of phosphinothricin and paromomycin for the selection of transgenic plants of spelt wheat. Methods. Shoot apical meristem culture, mature embryo culture, Agrobacterium-mediated genetic transformation. Results. Isolation and cultivation of shoot apical meristems of seedlings from three spelt genotypes and mature embryos from three other genotypes were carried out. A high frequency (from 80 to 100 %) of callus induction from explants was observed. It was shown that the addition of 5 mg/l of phosphinothricin or 100 mg/l of paromomycin to the culture medium almost completely inhibited plant regeneration compared to the control. After Agrobacterium-mediated genetic transformation of calli with a vector containing the phosphinothricin-N-acetyltransferase gene, regeneration of spelt shoots for one genotype was observed on a selective medium with 5 mg/l phosphinothricin. Conclusions. The selective concentrations of herbicide and antibiotic for obtaining transgenic spelt wheat plants with the corresponding marker genes are 5 mg/l for phosphinothricin and 100 mg/l for paromomycin.

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