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Obtaining of wheat (Triticum aestivum L.) lines with yeast genes of trehalose biosynthesis

Kvasko A.Yu., Isayenkov S.V., Dmytruk K.V., Sibirny A.A., Blume Ya.B., Yemets A.I.

SUMMARY. Yeast (Saccharomyces cerevisiae) trehalose biosynthesis genes (TPS1 and TPS2) were transferred into genomes of several common wheat varieties using two methods of Agrobacterium-mediated transformation (in vitro and in planta) to enhance their drought tolerance. For this purpose, vectors pBract214-TPS1 and pBract214-TPS2 were constructed using Gateway-cloning technique. Both vectors contained TPS1 and TPS2 genes under the control of the constitutive maize ubiquitin promoter (PUbi) and hygromycin-phosphotransferase (hpt) selec-table marker gene. Three-five days callus obtained from wheat immature embryos was used as explants for the transformation in vitro. Selection of transgenic plants was carried out on nutrient medium supplemented with 30 mg/L hygromycin (as selective agent).  Seeds of wheat (transgenic generation T1) were obtained after in planta method of transformation. Integration and presence of yeast  genes in wheat genomic DNA isolated from transgenic plants were confirmed by PCR analysis using primers specific to TPS1 and TPS2 genes.

Key words: trehalose, yeast genes of trehalose TPS1, TPS2, genetic transformation, Agrobacterium tumefaciens, in vitro, in planta, Triticum aestivum

Tsitologiya i Genetika 2020, vol. 54, no. 4, pp. 3-14

1. Institute of Food Biotechnology and Genomics NAS of Ukraine, Ukraine 04123, Kyiv 123, Osipovskogo st. 2a
2. Іnstitute of Cell Biology NAS of Ukraine, Ukraine, 79005, Lviv, Dragomanova st., 14/16

E-mail: kvasko.anna ukr.net, yemets.alla nas.gov.ua

Kvasko A.Yu., Isayenkov S.V., Dmytruk K.V., Sibirny A.A., Blume Ya.B., Yemets A.I. Obtaining of wheat (Triticum aestivum L.) lines with yeast genes of trehalose biosynthesis, Tsitol Genet., 2020, vol. 54, no. 4, pp. 3-14.

In "Cytology and Genetics":
A. Yu. Kvasko, S. V. Isayenkov, K. V. Dmytruk, A. A. Sibirny, Ya. B. Blume & A. I. Yemets Obtaining Wheat (Triticum aestivum L.) Lines with Yeast Genes for Trehalose Biosynthesis, Cytol Genet., 2020, vol. 54, no. 4, pp. 283–292
DOI: 10.3103/S0095452720040088

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