Orthogonal experimental design was adopted to tissue culture in banana (Musa acuminata Colla cv. Williams). Effects of BAP, NAA, IBA and different explants on explant direct budding were investigated in banana by using orthogonal design [L27 (313)]. Statistic analyses indicated that the main factor which mostly influenced the results was explants. BAP was the second factor, and both NAA and IBA the third. The results showed that the optimal medium was MS basal medium supplemented without the three growth regulators，and the best explant for the medium was the apical meristem of shoots．