This study was conducted to establish the maternal haploid method for the practical breeding of M tabacum using the interspeciflc hybridization between M tabacum and N. aflicana. The frequency of surviving seedling per seed capsule of interspecific hybridization was 4.15. Among them, the frequencies of maternal haploid and hybrid were 1.20 and 2.95, respectively. The chromosome numbers of n=24 for maternal haploid and 2n=47 for hybrid were identified in surviving seedling from interspecific hybridiztion. Except the chromosome number, distinguishable morphological differences of material haploid from hybrid were observed at seedling stage.