The invention discloses a rapid propagation method of culturing medical anoectochilus roxburghii tissue through which seedling is formed in only one step. The invention mainly consists of the flowing steps: firstly, culture medium is prepared by three steps of preparation, prepackaging and sterilization; secondly, explant is treated and inoculated, which step has two conditions: when a natural growth plant is used as an explant, the step consists of three steps of cleaning, sterilizing and inoculation, and when a tissue culture sterilized seedling is used as an explant, the sterilized inoculation is directly carried out; thirdly, cultivation; and fourthly, transplantation. The invention uses the middle stem of an anoectochilus roxburghii plant, which is produced by subculture, as an explant. A stem section which contains a stem node is inoculated on culture medium under sterile condition. The results show that after the optimum culture medium is cultured for four months, the average seedling number of each explant is 4.8; the average net increased number of the leaves of each plant is 3.6; the average increased root number is 2.5 and the average net increased fresh weight is 525.6mg. The growing condition of the tissue culture seedling is quite good after being transplanted to substrate. The survival rate of plants achieves to above 95 percent after three months. The invention can greatly shorten the tissue culture seedling time of anoectochilus roxburghii, significantly improve the culture efficiency and reduce the seedling cost. The invention can be used as an important method of producing anoectochilus roxburghii seedlings.