Effect of Genotype, Stress Pre-Treatment and Culture Medium on plant Regeneration of Barley Microspores Via Anther Culture

Doubled haploid production is one of the best breeding methods to produce homozygous (pure) lines. Microspore culture technique is the most efficient method for production of doubled haploid plants. This research was carried out to study the effect of genotypes, stress pre-treatments and culture media on the callus induction and plant regeneration from microspores of anthers in barley (Hordeum vulgare L.). Anthers were isolated from six barley genotypes including F3 hybrids: 21070, 21082, 21095, 21076, 21063, and cultivar Igri). Anthers were placed on the solid FHG medium without hormone or containing IAA (2 mgl-1) and BAP (1 mgl-1) or 2.4-D (2 mgl-1) and Kin (0.5 mgl-1). In addition, the effect of cold stress alone and the combination of cold and starvation treatments were investigated on callus induction and regeneration frequency. According to the results obtained, the highest number of callus formation and regeneration percentage was induced in genotype Igri when FHG medium supplemented by IAA (2 mgl-1) and BAP (1 mgl-1) without any pre-treatment was applied. The results showed that cultured anthers in genotype 21082 induced the highest green regenerates when pre-treated with cold stress for 28 days cultured on FHG medium supplemented by 2.4-D (2 mgl-1) and Kin (0.5 mgl-1). In conclusion, genotypes tested can be categorized to three groups including responsive (Igri), low–responsive (21082-21076) and recalcitrant (the rest genotypes).