Abstract: Multi-hand parallel transplantation can effectively improve the efficiency of vaccination, but the majority of transplanting hands based on dimension-positioning are complex, which causes operation space of transplanting hand not enough when several transplanting hands are working at the same time. Meanwhile, the ability is poor for adapting individual differences of tissue culture seedlings, some tissue culture seedlings will be held unstably or be damaged by transplanting hand in the process of holding. Based on those issues, a tissue culture seedlings transplanting hand was designed in this research, which used a negative pressure air flow to complete the guidance and hold of potato tissue culture seedlings, to strengthen the adaptability for individual differences of tissue culture seedlings, to achieve flexible hold. The transplanting hand for tissue culture seedlings mainly consists of three parts, including the sucker, vacuum generating device, and negative pressure detection part. The sucker is the key part of transplanting hand, which decides the holding and guide effect of transplanting hand for tissue culture seedlings. In this paper, design and experiments were both mainly focused on the guide and control performance of sucker. The process that sucker holds a tissue culture seedling can be divided into three phases: guide and hold, stem section cutting, and stem section inserting. In guide and hold phase, the transplanting hand guide tissue culture seedlings of deviation from the holding position to mouth of sucker by negative pressure airflow. In stem section cutting phase and stem section inserting phase, transplantinged hands need to be stable to hold tissue culture seedlings, to ensure effect of stem section cutting and inserting. At the same time, sucker of transplanted hands should avoid tissue culture seedlings from damage. According to the above requirements of transplanting hand, first, the three-dimensional modeling was established based on work for sucker of transplanting hand and fluid simulation analysis was carried out based on Fluent software for shape of sucker of transplanting hand with guide wings. The shape of sucker of the transplanting hand was round tube with the outer diameter 5 mm, and inner diameter 4 mm, and the angle was set 90° for guide wings in the front of round tube, the distance was 10 mm between two guide wing vertex. In order to test the ability of accommodating tissue culture seedlings deviation by negative pressure air flow guide for transplanting hand, orthogonal experiment was carried out, the results showed that potato tissue culture seedlings of average diameter was 1.23 ± 0.21 mm, the radius of transplanting hand that guided tissue culture seedlings successfully was 4 mm when the vacuum of sucker was 8.6 × 10?3 MPa. The process of vaccination should avoid the direction of transplanting hand movement and direction of tissue culture bending was vertical by adjusting three transplanting hand operation sequence in order to reduce the interference between the two. In order to test the hold performance of the transplanting hand, experiments of stem section cutting and stem section inserting were carried out under the condition of the vacuum of tube of 8.6×10?3 MPa，the results showed that the straight speed of cutter was 0.03 m/s when the rotate speed of cutter of thickness was 0.3 mm and diameter was 45 mm is 7 r/s. When stem diameter of stem section of tissue culture seedlings was (0.7 ± 0.1), (1.1 ± 0.1), (1.5 ± 0.1) mm and the inserting speed was 0.01 m/s, the success rate of stem inserting was 100%.