Anther Culture Derived Homozygous Lines in Hordeum bulbosum

The production of homozygous doubled haploids via anther culture was investigated in the self-incompatible diploid species Hordeum bulbosum. Anthers from three accessions (GBC77, GBC752 and GBC753) were cultured on FHG media using IAA or three levels (1, 50 and 100 mg/l) of the auxin phenylacetic acid (PAA). Four green plants and 63 albino plants were obtained from a total of 1620 plated anthers (540/accession). The best mean anther response, number of embryogenic calli and regenerated plants were obtained with 50 mg/1 PAA. Three of the four green plants survived to maturity and, based on root-tip squashes stained with feulgen, all had 14 chromosomes like the anther donor parent. These anther culture-derived plants contained only some of the parental DNA bands, as observed by PCR analysis, indicating that they are of gametic origin rather than arising from somatic parental tissue. This is the first report of homozygous lines produced from this self-incompatible species.