Genetic Diversity of Persian Clover Populations Using Semi-Random Markers

Field crop landraces are valuable genetic sources. Twenty populations of Persian clover (Trifolium resupinatum L.) collected from different areas of Iran were used in this study. DNA extractions were carried out using minipreparation method with equal amount of leaves from 30 plants of each population. DNA samples from 20 clover populations were evaluated using semi-random (ISJ) markers. Ten primers out of 30 which used IT (intron-targeting) and ET (exon-targeting) primers produced repeatable bands. Cluster analysis was conducted using NTSYS software and UPGMA method based on Jaccard's similarity matrix. Primers totally produced 111 bands, of which 93 bands (%93) were polymorphic among clover genotypes. The greatest and least amplification fragments belonged to IT15-31 and ET18-4 primers, respectively. Average band number per primer was estimated 11.1 bands. Furthermore, IT primers produced more polymorphic DNA fragments with higher resolution. Based on cluster analysis and cutting dendrogram in 0.8 similarity coefficients, clover populations were divided into five groups in which Kazerun and Kermanshahi (1) individually formed a separate cluster. According to similarity matrix, the least similarity (%42) belonged to Alvijan and Kazerun and the highest similarity belonged to Chegeni and Haftchin Hamedan. Clustering based on semi-specific PCR method almost substantiated the grouping based on geographical origin. Considering the results, it is concluded that PCR-based semi-random marker technique can be used for genetic diversity study of Persian clover as well as discrimination of its cultivars.