Evaluating the efficiency of DNA Barcode rbcL for detection of genetic relationships between four Moringa spp. genotypes

Abstract

Alterations in the DNA sequences are very useful for the unique markers development, which could be utilized as a barcoding of DNA for different plants. The barcoding of DNA is used for identification of living organisms and provides extra and integral support to identify the morpho-species, because it characterized by a reproducible, rapid and economic tool. This study was conducted to analyze the phylogenetic relationships between two species of the genus Moringa; M. oleifera and M. Peregrina, using DNA barcodes. In Egypt only two Moringa species (M. oleifera and M. Peregrina) were cultivated. These two species (four Moringa genotypes) were commonly used in the traditional medicine. To authenticate the different genotypes, rbcL regions were evaluated. DNA barcode regions were amplified using universal primers. The length of rbcL barcoding region varied between 638 bp in M. oleifera (II) collected from El-Suis governorate (Egypt) to 663 bp in M. peregrina from Giza (III), with a mean of 649.75 bp. In all investigated species, the percentage of GC content found to be nearly 44.01% and the span aligned sequences around 638-663 nucleotides. The number of variable sites within the sequences of the four Moringa genotypes was found to be 46 sites (11 transitions, 15 transversions and 20 indel). A phylogentic tree based on rbcL barcoding was constructed. Three clusters were shown, the first contained Moringa oleifera (R2) and the second contained Moringa oleifera (R1) and Moringa peregrina (R3). While the third cluster contained Moringa peregrina (R4). Several studies have reported similar observations. The results demonstrate that the analysis of these DNA barcode sequences is a reliable method for distinguishing between four Moringa genotypes.