Focusing on the problem of requiring a large number of labors for transporting culture bottles during the micropropagation period of tissue culture plantlets, a kind of automatic device for lining up culture bottles was designed. The automatic device transported bottles by using two conveyer belts which had opposite motion directions, and guided, assembled and separated bottles by using guiding board, guiding block and separating block respectively. The main factors which influenced sequencing speed and reliability were guiding belt speed v1, separating belt speed v2, dip angle α of guiding board and wedge angle β of guiding block. The sequencing speed and reliability were satisfactory with v1= 0.25 m/s, v2=0.4 m/s, α=47°and β=35°. Experiment results showed that the device could sequence automatically different size and shape bottles which were used frequently in plant tissue culture production and the sequencing reliability was more than 94%. The sequencing speed was nearly equal and the reliability was different while lining up bottles with same diameter and different shapes, because the bottles with less mass and lower center of gravity were easier to be lined up. Bigger bottles were easier to be sequenced than smaller ones while lining up bottles with same shape and different diameters. 