Investigation of genetic differences between chickpea blight agent ascochyta rabiei isolates with iPBS and SCoT DNA markers

The causative agent Ascochyta rabiei causes significant damage to chickpea plants. In this study, diseased chickpea plants obtained from different locations in Ankara and Çankırı provinces were used. Isolations from diseased plants collected from a total of 7 different locations were performed on chickpea flour agar medium, which is the most suitable medium for pathogen sporulation. The genomic DNA of the isolates was extracted using a CTAB-based method. The intra-species genetic diversity among the obtained A. rabiei isolates was investigated using the iPBS (inter-Priming Binding Sites) and SCoT (Start Codon Targeted) DNA marker systems. In preliminary work, the SCoT primers (13, 28, 29, 32) that were expected to provide polymorphism in distinguishing groups and evaluating genetic differences within groups were selected and used to detect intra-species differences among all isolates. The number of polymorphic bands amplified by iPBS primers ranged from 11 to 15, while the number of SCoT markers ranged from 7 to 13. The average number of amplified PCR products for iPBS and SCoT marker systems was found to be 10, 12, 13, and 11, respectively. However, no intra-species polymorphism was detected among the isolates using the iPBS and SCoT marker systems. It was concluded that the iPBS and SCoT marker systems are not suitable DNA marker systems for intra-species differentiation of A. rabiei isolates and further investigation with larger pathogen populations is necessary.