Gymnadenia conopsea,an alpine Orchidaceae plant,was one of the widely used Tibetan traditional medicines.In this study,we sequenced total 105 expressed sequence tags(ESTs)from a full-length cDNA expression library constructed by the Oligo-capping technique.The further bioinformatic analyses suggested that the 65 represented unique sequences showed high homology to previously identified genes in other plants: 30 sequences matched to other uncharacterized expressed sequence tags(ESTs),and 10sequences showed no good matches to available sequences in DNA databases.Gene ontology annotation by InterProScan indicated that many of these cDNAs(7 percent)have no known molecular functions and may be unique to G.conopsea.Fifty-five ESTs with matched proteins were involved in a series of diverse functions,in which molecular function such as"binding"(42.9percent)and"catalytic activity"(25.0 percent)were the most frequent functions of the cDNAs.This eDNA library provided a critical basis for further investigation of functional genes expression under cold stress in this alpine species.In addition,13ESTs-based polymerase chain reaction(PCR)primers were designed and can also be used for genotypic identification and for the genetic diversity analysis of G.conopsea and its closely related species.