The invention discloses a method for novel variety culture and rapid tissue culture propagation of paphiopedilum and relates to the technical field of breeding and tissue culture of paphiopedilum. Themethod for novel variety culture and rapid tissue culture propagation of paphiopedilum comprises the following steps: performing artificial hybridization pollination, performing sterile seed sowing and seed germination, strengthening seedlings, promoting rooting, and transplanting test tube seedlings. The method is good in operability, high in application value, high in hybridization success rate, and very effective for excellent cross parents of paphiopedilum in flowering asynchronism; in the rapid tissue culture propagation process, a unique seed germination culture medium and a seedling strengthening rooting culture medium are used, so that the germination rate of paphiopedilum hybridization seeds is increased, seedlings can grow rapidly, and the seedlings are good in quality. The method for novel variety culture and rapid tissue culture propagation of paphiopedilum, which is disclosed by the invention, has the characteristics of being good in operability, high in application value, high in hybridization success rate, and the like, the problems that conventional novel variety culture and rapid tissue culture propagation process of paphiopedilum is low in germination rate, hardin seedling growth, long in propagation cycle, low in propagation coefficient, large in propagation difficulty, and the like, are solved, an effective way is provided for production of paphiopedilum seedlings, and industrial planting can be achieved.