The Effect of Sugars and Growth Regulators on Embryoid Formation and Plant Regeneration from Barley Anther Culture

Abstract

AbstractThe influence of carbon source and growth regulator composition in induction medium on anther culture response was investigated using spring barley genotypes. Anthers were cultured on BAC3, Ficoll‐containing medium, supplemented with one of the following carbohydrates: sucrose, maltose, cellobiosc and melibiose (6 % w/v). The use of either maltose or cellobiose resulted in a significantly higher anther response, calli and/or embryoid production and green plant regeneration compared to the incubation of anthers on a medium containing sucrose. Contrary to these results, the replacement of sucrose by melibiose in BAO medium, drastically reduced the efficiency of anther culture. As an average for the three genotypes tested, the frequency of green plants per 100 anthers plated was 9‐ to 22‐fold higher on medium supplemented with sucrose or cellobiose than on medium containing melibiose as a sole carbohydrate.Among the growth regulators tested, the combination of auxin NAA (2 mg/l) and cytokinin BAP (1 mg/1) performed much better than the employment of auxin 2,4‐D combined either with zeatin riboside or BAP as cytokinins. The beneficial effect of medium supplemented with NAA and BAP was associated with better embryoid formation compared to the other growth regulator combinations tested. The hormone‐free combination gave a similar anther response and production of calli as any medium supplemented with growth regulators, but the regeneration capacity of calli produced on hormone‐free medium was much lower, resulting in the drastic reduction of the number of both total and green regenerants.