Efficient production of androgenic doubled-haploid mutants in barley by the application of sodium azide to anther and microspore cultures

The aim of this study was to establish a protocol for an efficient production of agronomical and/or physiological mutants from model (cvs. Igri and Cobra) and low-androgenic-responding (cv. Volga) cultivars of barley through the application of a mutagenic agent, sodium azide, to anthers and isolated microspores cultured in vitro. This technology offers the possibilities of screening for recessive mutants in the first generation, selecting for novel genotypes from very large haploid populations, avoiding chimerism and rapidly fixing selected genotypes as fertile true breeding lines. The mutagenic treatment, 10(-3)-10(-5) M sodium azide, was applied during the anther induction pre-treatment or immediately after the microspore isolation procedure. Out of 616 M-2 doubled-haploid lines characterised under field conditions, a total of 63 morphological and developmental independent mutant lines were identified. The percentage of M-2 doubled-haploid lines carrying mutations per line analysed was 3.8% when 10(-4) M sodium azide was applied to anthers from the low-responding cv. Volga; this increased to 8.6% and 15.6% when 10(-5) and 10(-4) M sodium azide were applied to freshly isolated microspores from model cultivars.