摘要
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.
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.
