Comparison of anther and isolated microspore cultures in barley.: Effects of culture density and regeneration medium

A comparison of anther and microspore culture efficiency for green doubled haploid plant production was undertaken using 17 F-1 crosses with potential agronomic performance. Green doubled haploid plants were produced from all F-1 crosses by anther and microspore cultures, although there was a great variation among crosses. On average, anther culture resulted in a production of green plant twice that of isolated microspore culture (30 and 14 green plants/100 anthers, respectively). The effect of microspore culture density on green plant regeneration was studied with the cultivars Igri, Reinette and Hop which have a high, medium and low androgenic response. The highest number of dividing microspores was obtained at a density of 2.4 x 10(5) viable microspores/ml for the three cultivars. However, the optimal density for the percentage of embryos/dividing microspores and green plants/10(3) microspores depended on the cultivar. The highest number of green plants/10(3) microspores was produced at 1.2 x 10(5) viable microspores/ml for cv. Igri and 2.4 x 10(5) for cultivars Reinette and Hop. Percentage of green plants/total plants was raised when the culture density was increased up to 6.0 x 10(5) viable microspores/ml, especially for cv. Reinette. Six regeneration media differing in maltose concentration, organic nitrogen and type of auxin were assayed with embryos from cultivar Reinette. Media without organic nitrogen containing 31 g l(-1) maltose and the auxins IAA or NAA produced more vigorous green plants.