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IRAP-analysis of transgenic wheat plants with double-stranded RNA suppressor of the proline dehydrogenase gene

Morgun B.V., Dubrovna O.V.

 




SUMMARY. The level of polymorphism of DNA regions flanked by inverted LTR retrotransposon repeats in genetically modified wheat plants containing the double-stranded RNA suppressor of the proline dehydrogenase gene obtained by the method of Agrobacterium-mediated transformation in an in vitro culture has been analyzed by IRAP analysis. When using all high-performance primers for the retrotransposons Sukkula, Sabrina, Wham, Nikita and Wilma1, no DNA polymorphism was found in transgenic plants. Under the conditions of the experiment, we did not register the disappearance of amplicons in the DNA profiles of PCR, which may indicate the absence of rearrangements in the binding sites with the primer and in the studied loci flanked by LTR retrotransposons. No new amplicons appeared in the spectra of DNA amplification products, which indicates the inactivation of the transposition phenomenon of mobile genetic elements in transgenic plants with a double-stranded RNA suppressor of the proline dehydrogenase gene. To expand the spectrum of amplicons in the PCR products of the studied samples, the method of combining IRAP primers with various retrotransposons in one reaction was tested. The pairs of IRAP primers were selected experimentally, however, using these pairs of primers, no disappearance or appearance of new polymorphic fragments was detected. The absence of DNA polymorphism in transgenic plants with a double-stranded RNA suppressor of the proline dehydrogenase gene may be associated with the phenomenon of RNA interference, which suppresses the activity of retrotransposons.

Key words: Triticum aestivum, Agrobacterium-mediated transformation, retrotransposon, IRAP PCR

Tsitologiya i Genetika 2019, vol. 53, no. 5, pp. 46-55

E-mail: molgen icbge.org.ua, dubrovny ukr.net

Morgun B.V., Dubrovna O.V. IRAP-analysis of transgenic wheat plants with double-stranded RNA suppressor of the proline dehydrogenase gene, Tsitol Genet., 2019, vol. 53, no. 5, pp. 46-55.

In "Cytology and Genetics":
B. V. Morgun, O. V. Dubrovna IRAP Analysis of Transgenic Wheat Plants with a Double-Stranded RNA Suppressor of the Proline Dehydrogenase Gene, Cytol Genet., 2019, vol. 53, no. 5, pp. 384–391
DOI: 10.3103/S0095452719050116


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