The objective of this study was to analyze the genetic diversity using SSR marker and investigate the fatty acid composition of perilla (P. frutescens var. frutescens) germplasm. Genetic diversity among 95 accessions, which consisted of 29 weedy types and 66 landrace accessions, was evaluated based on 12 SSR markers carrying 91 alleles. The mean values of observed ($H_O$) and expected heterozygosities ($H_E$) were 0.574 and 0.640, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into two distinct groups, which were the landrace, moderate and weedy type, genetic distance (GD) value was 0.609. The physicochemical traits about crude oil contents and fatty acid compositions were analyzed using GC. Among tested germplasm, the total average oil contents (%) showed a range from 28.57 to 49.67 %. Five fatty acids and their contents in the crude oils are as follows: ${alpha}$-linolenic acid (41.12%-51.81%), linoleic acid (15.38%-16.43%), oleic acid (18.93%-27.28%), stearic acid (2.56%-4.01%), and palmitic acid (7.38%-10.77%). The average oil content of wild types was lower than landrace, and the oil content of middle genotype accessions was higher than other germplasm, but no significant variation between landrace and wild types was shown. Nevertheless, IT117174, landrace of Korea, was highest in crude oil content (47.11%) and linolenic acid composition (64.58%) among the used germplasm. These traits of the selected accessions will be helped for new functional plant breeding in perilla crop.