The amount of genetic variability of a species is essential for its survival and adaptation in different environments, and studies of genetic diversity using molecular markers are necessary to understand the genetic structure of a population and to orientate effective strategies of germplasm conservation. The aim of current study was to determine the SSR markers that can be used rapidly and reliably to evaluated the pepper of Bulgaria landraces, and applied the markers to assement of introduce genetic diversity of the pepper germplasm. We used 22 polymorphic microsatellite markers to analysis of genetic diversity within 61 pepper collection of Bulgaria landraces germplasm, all SSR primers pairs produced 80 polymorphic and reproducible amplification fragments. An average value of polymorphic information contents (PIC) were 0.334 with a range of 0.061 to 0.63. The mean values of observed (HO) and expected heterozygosity (HE) were 0.383 and 0.154, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into three distinct groups, which was the landrace, moderate and wilde type, genetic distance (GD) value was 0.540. An average day of flowering time was 53 days with a range of 45 to 60 days. The everage od fruit length and width were 9.38cm with a range 2.1 to 23.6cm, and 3.51cm with a range 0.6 to 8.9cm, respectively. Molecular data were complemented with morphological measurements according to the descriptor list for the pepper collection of Bulgaria landraces germplasm.