The invention discloses an asymbiotic germination culture method of paphiopedilum gratrixianum seeds. The asymbiotic germination culture method comprises the following steps: cleaning, disinfecting and sterilizing paphiopedilum gratrixianum pods; cutting the pods into two sections by using a pair of aseptic scissors on an aseptic operating stand; sowing seeds in a sterilized germination culture medium, performing dark culturing at 25 DEG C in a darkroom, and regarding the appearance of white protocorms as the sign of seed germination; after the seeds generate the white protocorms, calculatingthe germination rate of the paphiopedilum gratrixianum seeds; after the seeds germinate and form the protocorms, transferring the protocorms to a place with irradiation to continue culturing; and after the protocorms change to green completely, transferring the protocorms into a paphiopedilum gratrixianum protocorm transferring culture medium to continue irradiated culturing. The paphiopedilum gratrixianum seed germination culture medium takes Hyponex 1 as a basic culture medium to which sucrose, agar, activated carbon and coconut powder are separately added. The germination culture method ofthe paphiopedilum gratrixianum seeds and use of the germination culture medium can prompt germination of the paphiopedilum gratrixianum seeds, improve the germination rate of the seeds, lay the foundation for seedling emergence and reintroduction of the paphiopedilum gratrixianum seeds, and are favorable for development of paphiopedilum gratrixianum conservation work.