Abstract: According to principle of bionics and friction inoculation of tobacco Mosaic virus，a minimally invasive inoculating device was designed.The motor was used to drive 800 abrasive cloth strip，and abrasive cloth strip was used to perform minimally invasive friction on smoke seedling.In order to avoid serious damage to delicate smoke seedling，the width of abrasive cloth strip was determined by the test to be 3 mm，and the speed of minimally invasive roller was 70 r/min.In order to facilitate daily management of tobacco vaccine，a tobacco inoculating spraying fertilization machine was designed with functions of inoculation，fertilization and drug spraying.The inoculating spraying fertilization machine was composed of frame，minimally invasive mechanism，control system，drug supplier and walking system.The mobile phone APP developed based on Eclipse was used as upper computer and STM32 as lower computer.Transmission of control instructions between the upper and lower computer was realized via Bluetooth，and conversion of three functions of spraying，fertilization and inoculation as well as control of the advance and turn of all-in-one machine was realized.Experiment showed vaccination rate of that seedbed inoculation spraying fertilization machine reached 87.3%，and index was excellent.Compared with tobacco Mosaic virus artificial inoculation method applied in production，this all-in-one machine achieved multi-purpose，broke through limitation of traditional artificial inoculation method，was minimally invasive and more adaptable，and improved working efficiency to a great extent.