The invention discloses a preparation method of Fuzhuan tea golden flower fungus spore powder, which comprising the steps of: inoculating golden flower fungus to a CZG slant culture medium, culturing for 5days at a temperature of 26-30 DEG C to obtain a seed culture; adding sterile water in the seed culture, washing down golden flower fungus spores, fully oscillating to obtain a uniformly-dispersed spore suspension; inoculating the spore suspension on the culture medium, carrying out constant-temperature culture at 26-30 DEG C, covering with a sterile breathable sealing film before white hypha germinates, removing the sterile breathable sealing film when yellow hypha germinates, continuously culturing for 3-5 days to obtain a yeast seed; then carrying out enlargement to obtain mouldy bran; and crushing the mouldy bran and then sieving to obtain the Fuzhuan tea golden flower fungus spore powder. The artificially cultured Fuzhuan tea golden flower fungus spore powder can be used for extracting effective components of the golden flower fungus and inoculating fermented Fuzhuan tea; and the fermentation period can be shortened and the pollution of infectious microbe is reduced.