German chamomile, Matricaria recutita L., is a medicinal plant belonging to Asteraceae family, which is known as the "star among medicinal species" due to its long history of utilization, dating back to the time of the Egyptian pharaohs. This species is endemic to Iran and its status has been introduced as a low-risk plant (LS). Numerous studies on the chromosome numbers of different species of the genus Matricaria have shown that the use of molecular and cytogenetic approaches allows obtaining additional information about the structure and functional organization of the its genome to answer questions associated with the classification and evolution of different species and the selection of new Matricaria races. Karyotype analysis and comparison of chamomile cultivars and wild species allow us to reveal their species characteristics. In the studies conducted on different cultivars of M. recutita species, the chromosomal number of this species was reported as 2n= 4x= 36 and 2n= 2x= 18. Cytogenetic studies showed that the chromosomes of M. recutita have a small size and similar morphology and are metacentric in nature. At the same time, the karyotypes of chamomile species have not been well studied and still need special attention. In this study, the chromosomal numbers of four different populations of M. recutita species collected from different regions of Iran were determined and after measuring different chromosomal parameters and karyotype, their cytogenetic traits and the distance and proximity of the populations were determined using the measured traits.