Application of beta-glucuronidase transient expression for selection of maize genotypes competent to genetic transformation

Nitovska I.O., Abraimova O.Ye., Duplij V.P., Derkach K.V., Satarova T.M., Rudas V.A., Cherchel V.Yu., Dziubetskyi B.V., Morgun B.V.

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SUMMARY. Genetic transformation of inbred lines and maize hybrids F₁, registered in Ukraine, has been carried out. The study used a biolistic method for the genetic transformation of maize immature embryos that formed callus tissue, pAHC25 vector containing the genes of phosphinothricin-N-acetyltransferase (bar) and β-glucuronidase (uidA). As a result of the transformation of callus tissue of maize genotypes, resistant to phosphinothricin lines and regenerated plants were obtained. The activity of β-glucuronidase in herbicide-resistant calli was detected. The presence of bar gene in the calli DNA was demonstrated by PCR method. The rate of stable transformation ranged from 2,2 to 30 % depending on genotype. The relationship of transient expression results of β-glucuronidase gene and stable genetic transformation was observed. The proposed protocol for the genetic transformation of maize using a transient expression study of β-glucuronidase gene allows for a significant simplification of selection of competent to genetic transformation genotypes and creation of transgenic organisms with new traits.

Key words: Zea mays L., immature embryos, biolistic genetic transformation, bar, uidA

Tsitologiya i Genetika 2019, vol. 53, no. 6, pp. 15-25

E-mail: molgen icbge.org.ua

Nitovska I.O., Abraimova O.Ye., Duplij V.P., Derkach K.V., Satarova T.M., Rudas V.A., Cherchel V.Yu., Dziubetskyi B.V., Morgun B.V. Application of beta-glucuronidase transient expression for selection of maize genotypes competent to genetic transformation, Tsitol Genet., 2019, vol. 53, no. 6, pp. 15-25.

In "Cytology and Genetics":
I. O. Nitovska, O. Ye. Abraimova, V. P. Duplij, K. V. Derkach, T. M. Satarova, V. A. Rudas, V. Yu. Cherchel, B. V. Dziubetskyi, B. V. Morgun Application of Beta-Glucuronidase Transient Expression for Selection of Maize Genotypes Competent for Genetic Transformation, Cytol Genet., 2019, vol. 53, no. 6, pp. 451–458
DOI: 10.3103/S0095452719060082

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