Niger (Guizotia abyssinica (L.f) Cass) is an important oilseed crop in Ethiopia. The outcrossingrnand self-incompatibility nature of the crop make breeding works difficult. Thernapplication of anther culture in niger improvement is very important in developingrnhaploids or double haploids. Double haploids enable to increase the selection efficiencyrnof desirable traits in recurrent selection scheme or used in producing synthetic cultivars.rnIdentification of the best responsive genotypes and appropriate media for callus inductionrnare prerequisite to regenerate plants using anther culture technique that supports thernbreeding program. Anthers of four niger varieties namely Shambu, Kuyu, Esete andrnFogera were cultured into four different basal media (MS, B5, N6 and LS). All basalrnmedia were supplemented with 2 mg/l 2, 4-D + 0.3mg/l KN or 2 mg/l NAA + 0.3 mg/lrnKN. Identification of the appropriate stage of harvesting capitula and optimization of therntime duration of cold-pretreatment was undertaken using anthers of Shambu. Callusrninduction experiment was also conducted on MS, B5 and NN media to identify the typesrnof calli induced. Various experiments were carried out to optimize callus induction, shootrnregeneration and rooting of plantlets using different combinations and concentrations ofrnPGRs. Data such as callus induction percentage, types of induced calli and regenerationrnpercentage of shoots and roots were collected and analyzed. Capitula (buds) fully orrnslightly covered by sepal having whitish- green or greenish-yellow anthers were thernoptimum stages of harvesting capitula and 24 hours time duration was recommended forrncold pretreatment at 40C. ANOVA showed that genotypes and media significantly affectrncallus induction percentage and regeneration. The effect of the two factors, however, isrnindependent. The average callus induction percentage was highest in Shambu (29.4%)rnfollowed by Fogera (21.4%) in the same treatment. It was possible to distinguishrnembryogenic calli (EC) from non-embryogenic calli (NEC) by visual observation. NECrnwere identified by their friable morphology and amorphous texture, which wererndominantly developed in MS media. Embryogenic calli were mainly produced in NN andrnB5 media, which were distinguished by their variable texture and colour between the twornmedia. Among ten and nine different PGRs combinations and concentrations, 2 mg/l 2,4-rnD + 0.3 mg/l KN and 2 mg/l KN + 1mg/l IAA were found to be superior PGRsrncombinations for callus induction and for shoot regeneration respectively. It was alsornfound that 0.5mg/l IBA was the better PGR for rooting. Regenerants of Shambu seem tornbe having better survival rate than the variety Fogera either in the growth room or glassrnhouse.rnKey Words / phrases: Guizotia abyssinica (L.f) Cass., anther culture, embryogenic callus, nonembryogenicrncallus, basal media, plant growth regulators.