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

Abstract

A comparison of anther and microspore culture efficiency for green doubled haploid plant production was undertaken using 17 F1 crosses with potential agronomic performance. Green doubled haploid plants were produced from all F1 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 105 viable microspores/ml for the three cultivars. However, the optimal density for the percentage of embryos/dividing microspores and green plants/103 microspores depended on the cultivar. The highest number of green plants/103 microspores was produced at 1.2 x 105 viable microspores/ml for cv. Igri and 2.4 x 105 for cultivars Reinette and Hop. Percentage of green plants/total plants was raised when the culture density was increased up to 6.0 x 105 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.