In the wild,the populations of Paphiopedilum are facing great threat because of habitat destruction and commercial demand for their beautiful and unique flowers.Mycorrhizal fungi are essential for orchid seed germination and seedling development in nature condition.To investigate mycorrhizal diversity and specificity in Paphiopedilum and use symbiosis technology for conservation and application of these orchids,we collected limited number of the root samples of P.malipoense and P.micranthum were from the field in Anlong,South Guizhou.The roots of P.hirsutissimum and P.dianthum were sampled from Guangxi Yachang Orchids Nature Reserve.Both of these places are karst land feature.The root samples of P.purpuratum were collected from the wild in Huizhou adjacent to Shenzhen,which is non-karst land feature.Sixteen isolates of Rhizoctonia-like fungi were isolated from the root pelotons of five mature orchids speices and identified by morphological and molecular characteristics.All these strains were belonged to Tulasnella (anamorph: Epulorhiza).Based on the phylogenetic analysis of the ITS-5.8S rDNA,thirteen Tulasnella isolates were separated into six groups.The fungi isolated from the roots of P.hirsutissimum,P.dianthum and P.malipoense shared group A and B,and two isolates came from P.dianthum belonged to group F.The strains associated with P.purpuratum were divided into group D and E,and only one fungi isolated from P.micranthum belonged to group C.These results showed that Paphiopedilum orchids had high specificity with Tulasnella fungi and were associated with various Tulasnella fungal taxa.In addition,the species in common habitats,like P.hirsutissimum,P.dianthum and P.malipoense,shared the same fungal taxa and P.purpuratum from distinctive habitat was associated with special Tulasnella fungal species.