Gene expression analysis is one of the important stages of modern molecular biology and increase a better understanding of complex biological processes. Real time PCR is the most precise method to evaluate the pattern of gene expression. Use of reference genes is the best method to normalize the data obtained from the mRNA molecule. Beta-carotene pathway from the geranyl-geranyl-pp which includes four different enzymes has not been identified in pomegranate. On the other hand a proper reference gene as a normalizer in real time PCR has not been introduced for this plant. For this purpose, the RNA was extracted using a CTAB based method from the different tissues of the pomegranate (leaf, flower, stem, seed and peel) and cDNA was synthetized. The stability of eight reference genes including rps9, 18s, actin, ubq, pal, cyp, cpr and ef1 was evaluated in the qPCR reaction and finally the beta-carotene pathway genes were partially sequenced and their expression was analyzed using the introduced reference gene. The results of the electrophoresis and the spectrophotometry demonstrated that the RNAs from the different tissues were extracted with high quality and quantity. Analysis of the eight reference genes in qPCR by the Norm Finder software revealed that the least (0.0246) and the most (0.2472) standard deviation (SD) is related to the cyp and 18s , respectively. In leaf and seed ubq , flower cpr , stem cyp and peel rps9 showed more expression stability . Mean comparison showed that the expression of lyc , psy and zdc genes was significant in the seed tissue. ) zdc ? psy ? lyc) . In the peel tissue, it showed that the expression of psy and lyc , zdc and lyc was not significant, while the expression of psy and zdc was significant ( psy ? lyc? zdc). The mean comparison also showed that the expression of three genes in the Shirin Poost Siyah Yazd genotype was not significant, while their expression in Qomi Poost Qermez and Torsh Poost Sefid Ryjab were significant ( lyc ? psy ? zdc ). ). Cyp was introduced as a stable internal reference gene in pomegranate. Lycopene pigment involved in the production of color, therefore, peels color is probably associated with lyc gene expression. The expression of lyc at the end of the pathway is more in the peel tissue, therefore, it can be concluded that the beta-carotene synthesis in the peel tissue is more than seed. Keywords: pomegranate, RNA extraction, qPCR, Carotenoid, gene expression