DNA Barcode: The Genetic Blueprint for Identity and Diversity of Phyllanthus amarus Schum. et. Thonn

Abstract

Since the identification of the plant species is an important event, the present work was carried out to identify the medicinal plant Phyllanthus amarus taking the genetic variability among the species into consideration. The chloroplast genomic DNA was isolated and tested for quantity and quality. 2 μl of the template DNA, master mix, molecular-grade water and the forward and reverse primer were added and the final volume was made to 50 μl, and the amplification was performed in a gradient thermo-cycler and tested for amplification. The product was purified and sequenced and the sequence was submitted to GenBank. The sequence was subjected to multiple sequence alignment, and the phylogenic tree was constructed using MEGA 4 bioinformatics tools. The genomic DNA of the chloroplast was isolated, the highly conserved non-coding intron region of t-RNA L of the chloroplast was amplified, the amplified fragment was sequenced and the sequence in FASTA format was subjected to Multiple Sequence Alignment. The sequence showed no homology with any other sequence reported in the GenBank and this was the first highly conserved non-coding intron sequence of the plant Phyllanthus amarus that was deposited and the phylogenic tree was constructed. The identification of plant species based on the morphology is acceptable to an extent but the genetic blueprint gives you the exact information for the identification of the plant species. DNA barcode is a standardized and cost-effective molecular identification system used for plant identification. Hence the medicinally valuable plants needs the accurate identification among the species belonging to the same genus.