Abstract: Disclosed is a biomechanical measuring technical method for maize seed radicle and coleorhiza separation, which is characterized by comprising the following operations: (1) seed sample preparation; (2) anterior tissue cutting; (3) radicle and coleorhiza separation; (4) coleorhiza sample acquisition; (5) coleorhiza sample fixation; (6) puncture force measurement; (7) information storage and analysis. The operation (3) includes the development of a maize radicle and coleorhiza separation device, and the operation (5) includes the development of a maize coleorhiza sample carrier. The present disclosure has the beneficial effects of providing direct biomechanical evidence for the research on the coleorhiza weakening regulation and control mechanism of the maize seed germination, and simultaneously providing reference for measuring the coleorhiza weakening biological force of the gramineous plant seeds.