TITLE:
DNA Fingerprinting of Essential Commercialized Medicinal Plants from Pakistan
AUTHORS:
Waqar Ahmad, Khushi Muhammad, Altaf Hussain, Habib Ahmad, Khalid Kahn, Iqbal Ahmed Qarshi, Kamran Iqbal Shinwari, Muhammad Shahid Nadeem, Youxiong Que, Ayaz Khan, Javed Iqbal
KEYWORDS:
RAPD, Decamers, Medicinal Plants, DNA Fingerprinting
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.8 No.9,
August
9,
2017
ABSTRACT: Development of fingerprints based on DNA markers is
necessary for proper identification and standardization of plant species. These
techniques are widely used to develop an unquestionable method of plant
identification to protect the patents and quality control for industry. In this
study, fifteen commercially important medicinal plants of Pakistan were
collected from botanical garden of Qarshi Industries (Pvt.) Ltd, Pakistan. The
objective was to optimize the extraction of genomic DNA for use in a PCR-based
random amplified polymorphic DNA marker approach. The initial protocol used 60
decamers to amplify scorable amplicons; only nine markers produced significant
bands in genomic DNA of medicinal plants. These markers generated 51 bands
ranging between 250 and 1600 bp. The most important property of genomic markers
is polymorphism to enable specific identification; all the used markers showed
100% polymorphism across 15 different plants. Further, six decamers amplified
specific bands to reliably identify 8 species. The amplified bands were
arranged in a binary matrix and analyzed by DNAMAN version 5.2.2 statistical
software. A homology tree was constructed using binary data for nine markers,
and four major clusters/clades were observed. The Rose, Mentha and Stevia
accessions had shown clear clustering and grouped in major clusters/clads I, II and III respectively. Sixty decamers amplified 51
polymorphic loci in the genomes of 15 commercially
valuable accessions. Moreover clear phylogenetic construction was observed in
the generation of homolog tree. This protocol could therefore be useful to
provide a baseline to authenticate, identify and perform phylogenetic analysis
of important medicinal plants used in the Pakistani herbal medicine industry.