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Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon’s information Index (S.I.)indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.