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.

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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