Molecular research on the genetic diversity of Tunisian date palm (Phoenix dactylifera L.) using the random amplified microsatellite polymorphism (RAMPO) and amplified fragment length polymorphism (AFLP) methods

Abstract

The genetic diversity organization and evaluation of relationships within Tunisian date-palm cultivars were assessed using the random amplified microsatellite polymorphism (RAMPO) and amplified fragment length polymorphism (AFLP) methods. 18 combinations of random/ISSR primers and six AFLP primers combinations were tested with DNAs isolated from 40 date-palm cultivars. Our results show that using both markers systems, the Tunisian date-palm germplasm is characterized by a large and continuous genetic diversity. In addition, due to the greater number of markers per assay, the AFLP technique seems to be more informative than the RAMPO method. In fact, 186 and 428 polymorphic bands were detected using RAMPO and AFLP primers, respectively. Moreover, AFLP markers were found most polymorphic with the highest average PIC value (0.7) and marker index (50.54). In addition, independent as well as combined analyses of the cluster analyses of the RAMPO and AFLP fragments showed that cultivars are clustered independently from the sex of trees or else their geographical origin. On the other hand, based on Pearson and Spearman correlation between RAMPO and AFLP distance, matrices were positive and highly significant. This result indicates good congruence between these two molecular markers. The opportunity of the designed methods is discussed with the molecular characterization of genotypes in order to enhance the conservation and the improvement of the local date-palm germplasm.

Key words: Phoenix dactylifera, genetic diversity, random amplified microsatellite polymorphism (RAMPO), amplified fragment length polymorphism (AFLP), Tunisian.