Lines were developed from the two crosses, BY4 X NC 82 and BY 4 X BY 104, by bulk, backcross and anther-derived dihaploid methods of breeding , and their variations in major agronomic, chemical characters, heritability, and response to selection were compared. 1. The anther-derived dihaploid lines showed a greater genetic variability and heritability in characters investigated than the lines developed by the other two methods of breeding . However, those were Inferior In agronomic characters and yielded 15.4% less leaf tobacco than the lines developed by the bulk method. 2. The lines developed by bulk method were higher In the frequency of lines resistant to bacterial wilt and in the yield, but Its lines appeared later flowering. inferior leaf Quality, and higher total nitrogen contents than those developed by the backcross method of breeding. 3. In the population of the lines developed by the backcross method, bacterial wilt resistant lines were less frequent, but the lines with a good leaf quality were more frequent.