Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro

  • O. V. Dubrovna
  • L. V. Slivka
DOI: https://doi.org/10.7124/FEEO.v26.1264

Abstract

Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes. Methods. Agrobacterium-mediated transformation in culture in vitro. Results. The influence of the optical density of cells of the agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated using LBA4404 and AGL0 strains. It is shown that depending on the strain the most optimal is the concentration of agrobacteria 0.2-03 OD, duration of coculture for 2-3 days and the use of cefotaxime at a concentration of 250-500 mg/L. Conclusions. The optimal parameters for conducting Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes were selected.

Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures.

References

Hiei Y., Ishida Y., Komari T. Progress of cereal transformation technology mediated by Agrobacterium tumefaciens. Frontiers in Plant Science. 2014. Vol. 5. Р. 1–11. doi: 10.3389/fpls.2014.00628.

Dubrovna O.V., Morgun B.V. Current status of research of Agrobacterium-mediated transformation of wheat. Plant physiology and genetics. 2018. Vol. 50 (3). P. 187–217. [in Ukrainian]

Bhalla P., Ottenhof H., Singh M. Wheat transformation an update of recent progress. Euphytica. 2006. Vol. 149. P. 353–366. doi: 10.1007/s10681-006-9087-6.

Kumlehn J., Hensel G. Genetic transformation technology in the triticeae. Breeding Science. 2009. Vol. 59. P. 553–560. doi: 10.1270/jsbbs.59.553.

Opabode J. Agrobacterium-mediated transformation of plants: emerging factors that influence efficiency. Biotechnology and Molecular Biology. 2006. Vol. 1. P. 12–20.

Chauhan H., Khuran P. Use of doubled haploid technology for development of stable drought tolerant bread wheat (Triticum aestivum L.) transgenics. Plant Biotechnol. J. 2011. Vol. 9. P. 408–417. doi: 10.1111/j.1467-7652.2010.00561.

Сhugh A., Vikrant A., Mahalakshmi A., Khurana P. A novel approach for Agrobacterium-mediated germ line transformation of Indian bread wheat (Triticum aestivum) and Pasta wheat (Triticum durum). J. of Phytol. 2012. Vol. 4 (2). P. 22–29.

Bavol A.V., Dubrovna O.V., Lyalko I.I. Regeneration of plants from the explants of the top of wheat seedlings shoots. Bul. Ukr. Soc. Genet. Breeders. 2007. Vol. 5 (1–2). P. 3–10. [in Ukrainian]

Cheng M., Fry J., Pang S., Zhou H., Horinaka C., Duncan D., Conner T., Wan Y. Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol. 1997. Vol. 115. P. 971–980. doi: 10.1104/pp.115.3.971.

Sparks C., Doherty A., Jones H. Genetic transformation of wheat via Agrobacterium-mediated DNA delivery. Methods Mol Biol. 2014. Vol. 1099. Р. 235–250. doi.org/10.1007/978-1-62703-715-0_19.

Chen D., Dale P. A comparison of methods for delivering DNA to wheat: the application of wheat dwarf virus DNA to seeds with exposed apical meristems. Transgenic Res. 1992. Vol. 1. P. 93–100. doi: 10.1007/BF02513026.

Mahalakshmi A., Khurana P. Agrobacterium mediated gene delivery in various tissues and genotypes of wheat (Triticum aestivum L.). J Plant Biochem Biotechnol. 1995. Vol. 4. P. 55–59. doi: 10.1007/BF03262953.

Sharma V., Hansch R., Mendel J. Node-derived cultures with high-morphogenetic competence in barley and wheat. Plant Cell, Tissue and Organ Culture. 2007. Vol. 88 (1). P. 21–33.

Ahmad A., Zhong H., Wang W., Sticklen M. Shoot apical meristem: In vitro regeneration and morphogenesis in wheat (Triticum aestivum L.). In vitro cellular and development biology. 2002. Vol. 38 (2). P. 163–168. doi: 10.1079/IVP2001267.

Gorbatiuk I.R., Shcherbak N.L., Bannikova M.O., Velikozhon L.H., Kuchuk M.V., Morgun B.V. Obtaining herbicide phosphinotricin transgenic plants of wheat varieties Zymoyarka in vitro transformation. Plant physiol. genetics. 2016. Vol. 48 (1). P. 65–74. [in Ukrainian]

Sharma V, Hansch R, Mendel R, Schulze J. A highly efficient plant regeneration system through multiple shoot differentiation from commercial cultivars of barley (Hordeum vulgare L.) using meristematic shoot segments excised from germinated mature embryos. Plant Cell Rep. 2004. Vol. 23. P. 9–16.

Bavol A.V., Dubrovna O.V., Goncharuk O.M., Voronova S.S. Agrobacterium-mediated transformation of wheat using calli culture. Faktory eksper. evoliucii orhanizmiv. 2015. Vol. 15. P. 16–19. [in Ukrainian]

Rashid H., Afza A., Khan M., Chaudhry Z., Malik S. Effect of bacterial culture density and acetosyringone concentration on Agrobacterium mediated transformation in wheat. Pakistan Journal of Botany. 2010. Vol. 42. P. 4183–4189.

Vasil I.K. Molecular genetic improvement of cereals: transgenic wheat (Triticum aestivum L.). Plant Cell Reports. 2007.Vol. 26. P. 1133–1154. doi :10.1007/s00299-007-0338-3.

Dubrovna O.V., Bavol A.V., Zinchenko M.O., Goncharuk O.M. Effect of cefotaxime on morphogenesis in the culture of apical meristems and mature embryo of wheat. Physiol. Biochem. Cult. plants. 2012. Vol. 44 (3). P. 218–224. [in Ukrainian]