Camelina (Camelina sativa (L.) Crantz), an oilseed crop, belongs to the Brassicaceae family. Two unique features of camelina in comparison with the main oil crops are an adaptation to different environments and also its unique oil composition. The development of doubled haploid plants is one of the essential methods for crop improvement. The study of genetic diversity is an important step in planning crop breeding programmes. This research was conducted to evaluate the genetic variation of 81 camelina doubled haploid lines obtained from 15 crosses by inter simple sequence repeat (ISSR) markers. The total number of amplified bands was 243, of which 239 bands (98.3%) showed polymorphism. The percentage of polymorphic bands varied between 93.75 and 100. The size of the bands ranged from 50 to 1700 base pairs. The informative ISSRs were identified by estimating marker features: polymorphism information content, effective multiplex ratio, marker index and resolving power. Three markers had higher resolving power values (9.88, 8.5 and 7.46) and were the most informative markers to identify the lines. Cluster analysis based on the complete algorithm divided the lines into five groups, indicating relatively clear configuration from the geographic distribution patterns of the parents of the doubled haploid lines. Principal coordinate analysis classified the 81 camelina doubled haploid lines into six groups. The ISSR markers detected high polymorphism to reveal the genetic variation of camelina lines. The findings of this research, along with the characterization of biochemical traits of the lines, can improve breeding programmes achieve high-yielding camelina varieties with higher and better oil content.