The invention discloses a tissue cultural method of butterfly orchid. The method comprises the steps of seedling cultivation of proliferation nutrient solution which includes compound major elements, compound micro substances, inositol, 6-benzyl purine, 2,4-dichlorobenzene acetic acid, indolebutyric acid, mashed potatoes and saccharose; strong seedling cultivation of strong seedling culture solution which contains compound major elements, compound micro substances, inositol, peptone, activated carbon, citric acid, ammonium nitrate, nitrate of potash, mashed potatoes, mashed bananas and saccharose; adaptation cultivation of growing culture solution which comprises compound major elements, compound micro substances, inositol, indolebutyric acid, peptone, activated carbon, citric acid, ammonium nitrate, nitrate of potash, major element auxiliaries, mashed potatoes, mashed bananas and saccharose. A culture medium of butterfly orchid is also provided in the invention, and by using the culture medium, the germination rate is increased by 6%; the growing period of seedlings is shorten by 10 days; the survival rate of the seedlings is 100%; the growing period of the strong seedlings is shorten by 14 days.