Study of transgene expression in Triticum aestivum L. after Agrobacterium-mediated in planta transformation

  • N. A. Zhalii Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho Street, 148; National University of Food Technologies, Ukraine, 01033, Kyiv, Volodymyrska Street, 68
  • M. O. Bannikova Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho Street, 148
  • M. O. Plugatar STC «Institute of Biology and Medicine», Kyiv National Taras Shevchenko University, Ukraine, 01601, Kyiv, Volodymyrska Street, 64
  • L. H. Velikozhon Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho Street, 148
  • A. M. Taranenko Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho Street, 148
  • B. V. Morgun Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnoho Street, 148; STC «Institute of Biology and Medicine», Kyiv National Taras Shevchenko University, Ukraine, 01601, Kyiv, Volodymyrska Street, 64
DOI: https://doi.org/10.7124/visnyk.utgis.16.2.1053

Abstract

Aim. Detection of sequences of target transgenes nptII and bar in the genome of probable transformants of bread winter wheat Triticum aestivum L. cultivars Zymoiarka and Podolianka obtained as a result of Agrobacterium-mediated transformation in planta and determination of their expression level. Methods. Polymerase chain reaction (PCR) method was used independently and in combination with reverse transcription (RT-PCR), electrophoresis of DNA in agarose gel. Tolerance to the herbicide was evaluated in the physiological test. Results Through PCR analysis, the sequence of nptII transgene was detected in 30 samples of 145 analyzed, the frequency of transformation was 20.7 %. The sequence of the gene bar was observed in 85 experimental plants, and the frequency of transformation was 15.6 %. mRNAs of both transgenes were detected, indicating their transcriptional activity and stable expression. Conclusions PCR analysis allowed to detect nptII transgenic signal in 20.7 % of plants, while the presence of the bar gene was detected in 15.6 % of cases, indicating a higher efficiency of this genetic construct. The transcription is shown in all the specimens studied for both transgene. According to the results of the physiological test, 25 % of plants containing the gene bar showed resistance to the Basta® herbicide.
Keywords: genetically modified organisms, transgenic plants, biotechnological cultures, bread winter wheat, genetic engineering.

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