Sequence Analysis of nrDNA ITS Region in Paşa and Şah Apple (Malus × domestica) Genotypes

Abstract

In this study, phylogenetic analysis of Şah and Paşa apple genotypes was performed using nrDNA ITS sequences. After the plant leaves were brought to the laboratory, gDNAs were obtained by genomic DNA isolation method. PCR amplification was performed using primers ITS4 and ITS5A. ITS sequences of some apple and Rosaceae species were retrieved from NCBI, bioinformatics analyzes were made with Bioedit 7.2.3, Finch TV 1.4.0 and MEGA 6.0 programs and phylogenetic trees were constructed. In the study, ITS sequence length of the Paşa apple genotype was 656 bp, and the ITS sequence length of the Şah apple genotype was 649 bp. Only in the maximum likelihood phylogenetic tree constructed using sequences of apple genotypes, Paşa and Şah apple genotypes appeared in the same clade as Malus domestica cultivar="Casciana'' and Malus domestica cultivar="Rotella''. In the phylogenetic tree generated including other species belonging to the Rosaceae family; apple genotypes Pyrus, Cotoneaster, Crataegus, Sorbus, Eriobotrya and Prunus were detected in a same clade. Overall results clearly suggested that the ITS sequences were both suitable for differentiation between the selected genera and were compatible with previous phylogenetic studies.