The invention relates to the technical field of edible fungus cultivation, particularly to a method for applying cedar sawdust flammulina velutipes culture medium waste in agrocybe cylindracea cultivation. The method comprises the following steps of, firstly, preparing flammulina velutipes culture medium with cedar sawdust, and then utilizing the flammulina velutipes culture medium to cultivate flammulina velutipes; secondly, after the flammulina velutipes in the first step is harvested, obtaining flammulina velutipes culture medium waste, and preparing agrocybe cylindracea culture medium with the flammulina velutipes culture medium waste; thirdly, utilizing the agrocybe cylindracea culture medium obtained in the second step to cultivate agrocybe cylindracea. Through the method for applying the cedar sawdust flammulina velutipes culture medium waste in agrocybe cylindracea cultivation, the cultivated agrocybe cylindracea is not only rich in 17 amino acids such as asparaginic acid and glutamic acid, B vitamins as well as more than ten kinds of mineral trace elements, but also contains large amounts of anti-cancer polysaccharide, thereby obtaining high anti-cancer effects. Meanwhile, the cultivated agrocybe cylindracea has the advantages of being vigorous in growth, high in yield, white, tender and attractive in appearance, and fresh and crispy in taste.