The present experiments were conducted to investigate some of the factors affecting the number of haploids derived from anther culture of Nicotiana tabacum. Anther - donor plants grown under controlled environment room at 3$0^{circ}C$ yielded more haploid than room at 18, 25 and 26-22-18$^{circ}C$ in anther culture. Donor plants starved of fertilizer yielded more haploids as compared to those of the well fed with fertilizer in anther culture. Pretreatment of exercised flower bud at 5$^{circ}C$ was shown to be more effective in anther culture than pretreatment at 7 and 1$0^{circ}C$, and the optimum temperature and period of pretreatment were 4 or 6 days at 5$^{circ}C$.