Amsonia orientalis Decne. (Apocynaceae), is a rare and threatened plant species which is located only in a constricted area in northeast of Greece and northwest of Turkey in the world. Although phylogenetic analysis depending on nucleotide sequences of genes from different sources (nucleus, mitochondria and chloroplast) became a major tool for classification of plant species, there is still a big lack of information about A. orientalis in the international molecular data bases such as NCBI. In the current study, we phylogenetically analyzed three commonly used molecular markers (18S rDNA, 18S-28S rDNA-ITS region and trnL-F intergenic spacer) from A. orientalis samples collected from Turkey to determine the genetic diversity and also to question the systematic position of A. orientalis. As a result, A. orientalis samples clearly showed close relation with Alyxieae tribe rather than Vinceae. And this result brings the necessity to reconsider the morphological characters that have used to delimit the tribes of Rauvolfioideae.