Inhibition of Poly Phenol Oxidation during Genomic DNA Isolation and Morphological Variations in Berberis lycium Royle Genotypes

Abstract

Berberis lyceum is an economically and medicinally important genus distributed abundantly in the mountainous ranges of Pakistan. The color of hairs on the leaves of B. lyceum genotypes was pale to brown while the branch color was brown in all B. lyceum genotypes. Similarly, flower color was pale yellow while the plant height was ranged from 2.0 to 5.0 meters. Average length of lamina was 2.4 cm, average numbers of thorns per branch were 50.0 and average number of berries per branch was 18. Leaves tend to have comparatively high phenolic compounds that can interfere with DNA isolation. DNA-based markers are becoming popular for the identification of plants. Molecular genetic diversity studies of Berberidaceae family are almost lacking, which revealed the neglected status and the extent of the gap in the knowledge of this family. Mercaptoethanol being a reducing agent directly or indirectly causes damage to DNA. It is one of the limiting factors found in our study that can inhibit the oxidation process and at its low concentration brown viscous DNA pellet was extracted from fresh leaves while at high concentration intact whitish DNA pellet of good quality was observed that found suitable for PCR amplification. The DNA quantity and quality from dry leaves were quite low that was improved with the increase in the concentration of β 2-mercaptoethanol. It is concluded that a combination of 1% PVP and 0.01 to 0.06 % β mercaptoethanol was found satisfactory for extracting DNA from fresh and dry leaves of B. lyceum genotypes. Further, fresh and dry leaves were not able to exhibit any significant effects.