Molecular Genetic Diversity and Phylogenetic Analyses of Punica granatum L. Populations Revealed by ISSR Markers and Chloroplast (cpDNA) trnL-F Region

In this study, molecular genetic diversity and phylogenetic analysis of Turkish (Aegean region) Punica granatum L. populations was conducted based on ISSR-PCR and chloroplast DNA trnL-F sequences. Genomic DNA samples were isolated from fresh and green pomegranate leaves. Eight ISSR primers were used to determine genetic diversity between pomegranate populations. For the amplification of the cpDNA trnL-F region, trne and trnf primers were used in PCR. In the ISSR-PCR analysis, all gel images were examined and the presence and absence of polymorphic bands were scored. In ISSR-PCR, a total of 55 bands were obtained. Polymorphism rate approximately 78% were determined. UPGMA dengogram consist of two clades. For cpDNA trnL-F analysis, sequences were analysed using BioEdit and FinchTV programs. Both phylogenetic analyses and genetic distances were obtained using MEGA 6.0 program. Maximum likelihood and bootstrap trees were generated to determine genetic relationships between pomegranate populations. For populations, trnL-F sequences were detected between 357 and 384 bases. In addition, trnL-F sequences of some species of Lythraceae family from NCBI with pomegranate populations phylogenetic tree was constructed. Analysis of pomegranate populations based on ISSR-PCR and cpDNA trnL-F sequences revealed that the rate of polymorphism obtained in the ISSR-PCR results were higher than the results obtained from the trnL-F sequences.