Effect of trophic, osmotically active and solidifying components of nutrient medium on the direct embryognesis in spring barley anther culture in vitro

  • E. V. Belinskaya
  • P. G. Dulnyev


Aim. Ambryoidogenesis, or somatic embryogenesis, is known to be the most efficient mode of plant regeneration in plant cell, tissue and organ culture. The investigation was aimed to elucidate effects of trophic and osmogenic components of inductive medium on the frequency of direct embryoidogenesis in spring barley anther culture in vitro and to determine mechanism of morphogenesis improvement coursed by chemically modified starch D-5aM used as a gelling agent instead of agar. Methods. Anthers of DH-line with a high androgenetic capacity were inoculated on inductive media containing N6 macro-, MS micronutrients, organic supplements, maltose or mannitol (0.3 M) and solidified with agar or chemically modified starch. Results. A positive effect of combination of high maltose content and chemically modified starch on the induction and regeneration processes in spring barley anther culture in vitro was confirmed. It was also shown that mannitol didn’t keep any growth or development processes going in barley anther and embryo culture, but at the same time this substance had no toxic effect. Conclusions. In order to achieve a high frequency of induction in spring barley anther culture, it is necessary to use medium containing maltose – a low weight component both with trophic and with osmotic activity.

Keywords: Hordeum vulgare L., anther culture in vitro, mannitol, maltose, embryo formation, plant regeneration.


Bergen S., Wang M. Microspore regeneration as a tool for plant breeding. Acta Horticulture. 2002. No. 572. P. 51−57. doi: 10.17660/Acta Hortic.2002.572.5.

Sorvari S., Schider O. Influence of sucrose and melibiose on barley anther culture in starch media. Plant Breeding. 1987. Vol. 99, No. 2. P. 161–171. doi: 10111/j1439-0523.1387.tb01167.x.

Roberts-Oehlschlager S.L., Dunwell J.M. Barley anther culture: pretreatment on mannitol stimulates production of microspore-derived embryos. Plant Cell, Tissue and Organ Culture. 1990. Vol. 20, No. 3. P. 235−240. doi: 10.1007/BF00041887.

Bilynska O.V. Effect of low molecular weight osmogenic substances on the induction of embryogenesis in spring barley anther culture in vitro. Factors in experimental evolution of organisms. 2017. Vol. 20. P. 164−167 (in Ukrainian).

Bilynska O.V., Dulnyev P.G. Morphogenic effect and trophic capacities of chemically modified starch D-5aM in spring barley (Hordeum vulgare l.) anther culture in vitro and embryoculture. Factors in experimental evolution of organisms. 2015. Vol. 17. P. 107−111 (in Ukrainian).

Вilynska O.V. Effect of low molecular weight osmogenic substanses and solidifying agent of the medium on barley haploid production in anther culture in vitro. Plant Cells in vitro: Fundamentals and Applications II: Book of Abstracts of international conference (Vienna, Austria, 26−27 Jun 2017). Vienna: VISCEA, 2017. P. 38.

Belinskaya E.V. Inheritance of potential for in vitro androgenesis in spring barley. Сytology and Genetics. 2008. Vol. 42, No. 4. P. 237−245. doi: 10.3103/S009545270804004X.

Belinskaya E.V., Dulnyev P.G. Modified starch dkkmod as a component of nutrient medium for barley haploid production in anther culture in vitro. Physiology and biochemistry of cultivated plants. 2007. Vol. 39, No. 2. P. 136–143 (in Russian).

Murashige T., Skoog F. A revised medium for growth and bioassays with tobacco tissue cultures. Physiol. Plant. 1962. Vol. 15. P. 473–497. doi: 10/1111/j.1399-3054.1962.tb08052.x.

Bilynska O.V. Effect of solidifying and trophic carbohydrate components of nutrient medium on barley haploid induction in anther culture in vitro. Naukovyi visnyk of NUBiP of Ukraine. 2009. Is. 132. P. 17–24 (in Ukrainian).

Lersten N.R. Flowering plant embryology. Ames (USA): Blackwell Publishing, 2004. 212 p.