With the aim of investigation of the Cytotaxonomy of two species of Plantago genus from the family of Plantaginaceae, collected from the habitats of the northern provinces of Iran, the karyotype of 21 populations belonging to P. major L. and P. lanceolata L. is studied in the meristem cells of root tip using an image analysis system. The chromosomal dimensions such as the length of short arm and the length of whole chromosome are measured. The AR and CI statistics are also calculated after preparation, staining, and microscopic investigation of samples. Separation of chromosomes by photoshop software through Micromeasure software in at least 3 appropriate cells of mitotic metaphase observed. The populations are measured based on karyotype statistics including S%, TF%, A1, DRL, A2, VRC, karyotype formula and grouping the Stebbins for studied populations. The results of Principal Componet Analysis (PCA) show that two first main component totally indicates the 21.5% of total Variance. Dendrogram of cluster analysis using ward method, shows nicely separation of four cluster for twostudied species. In this research, the studied populations of P. major L. and P. lanceolata L. had the chromosomal number of 2n=2x= 12 and the basic chromosome number was equal to x= 6 in all populations and it was consistent with previous studies and this indicates the diploid species in Iran. Furthermore, according to the Levans Table, the chromosomes of studied populations were from the metacentric and submetacentric species.
