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Fiziol. rast. genet. 2020, vol. 52, no. 2, 128-139, doi: https://doi.org/10.15407/frg2020.02.128

PCR analyses of first-generation plants of Amaranthus caudatus L. after «floral-dip» genetic transformation

Yaroshko O.M.1, Morgun B.V.1,2, Velyko­zhon L.G.1,2, Gajdošova A.3, Andrushenko O.L.4, Kuchuk M.V.1

  1. Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine Akademika Zabolotnoho St., 148, Kyiv, 03143, Ukraine
  2. Institute of Plant Physiology and Genetics, National Academy of Sciences of Ukraine Vasylkivska St., 31/17, Kyiv, 03022, Ukraine
  3. Institute of Plant Genetics and Biotechnology PSBC Slovak Academy of Sciences Akademická St., 2, Nitra, 95007, Slovak Republic
  4. M.M. Gryshko National Botanical Garden, National Academy of Sciences of Ukraine, Kyiv, Ukraine Tymiriazievska St., 1, 01014, Kyiv, Ukraine

Amaranth is the C4-plant which used in various industries and is a promising object for genetic transformation. After «floral-dip» transformation of Helios and Karmin cultivars of Amaranthus caudatus L. using Agrobacterium rhizogenes A4 strain and A. tumefaciens GV3101 strain, transformed plants of the first generation were obtained. Gene vectors pCB125 and pCB131 contained a bar gene imparting resistance to commercial herbicide Basta. Seeds of Helios and Karmin cultivars of amaranth were obtained. The T-DNA transmission of strains A4 (pCB131) and GV3101 (pCB125) was evaluated by herbicide selection of amaranth plants. Tolerant plants were obtained for both cultivars after spraying with the herbicide. The percentage of A. w caudatus cv. Helios (vector pCB125) plants tolerant to the effect of the herbicide (160 mg/l) was 4.05 %, cv. Karmin (vector pCB125) — was 2.4 %. The percentage of tolerant A. w caudatus cv. Karmin (vector pCB131) plants was 0.6 %. The transgene presence of pCB125 gene vector (A. tumefaciens strain GV3101) and pCB131 vector (A. rhizogenes A4 strain) in amaranth tissue was analyzed. Integration of the bar gene and nos terminator in plants was confirmed by PCR analysis. The percentage of bar-positive plants for cv. Helios (gene vector pCB125) was 0.6 % of the total initial quantity of plants (12 positive plants of 2,000), and for cv. Karmin (gene vector pCB131) was 0.1 % of the total initial quantity of plants (2 plants of 2,000). The percentage of nos terminator-positive plants for cv. Helios was 0.15 % of the total initial quantity of plants (3 plants of 2,000).

Keywords: Amaranthus caudatus L., Agrobacterium, transformation, genetically modified plants

Fiziol. rast. genet.
2020, vol. 52, no. 2, 128-139

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