Abstract
Objective: The objective of this research was to develop the RAPD based SCAR marker for the correct identification of the Iris ensata Thunb. (I. ensata) plant from its adulterants.Methods: Five samples of I. ensata. from the different geographical area were used in this study. The plant genomic DNA was isolated with the CTAB method with some modification (as dried samples were also used). After that, polymorphism was checked with the help of the 10-mer random primers of OPAA and BG series. Then, the bands of interest were eluted and cloned into pGEMT easy vector for the sequencing. Finally, the sequence is used to develop the SCAR primers (Ir-f andIr-R) specific for I. ensata and the developed primers also validated with respect to the market samples.Results: A putative 580 bp sequence specific for Iris ensata was identified from the randomly amplified polymorphic DNA (RAPD) analysis. To overcome the main limitation of RAPD it has been converted into SCAR markers. So that, this specific band was then eluted, cloned and sequenced. After that, SCAR primers (Ir-F and Ir-R) were synthesized by using this sequence. For the validation of the synthesized SCAR primers, they were tested with respect to the market samples. The amplicon of 260 bp was produced by the SCAR primers in the authentic I. ensata but market samples did not produce any bands with the synthesized SCAR primers.Conclusion: The results of this study show a high level of polymorphism in the RAPD pattern of the different accessions of the plant. Furthermore, this study results in the successful development of the RAPD based SCAR marker for the identification of the I. ensata.
Highlights
In Iridaceae family, Iris is the major genus which has near about 230 species [1]
We developed reliable sequence characterized amplified region (SCAR) markers based on the amplified product of the randomly amplified polymorphic DNA (RAPD) primers for the identification of Iris ensata from its adulterants
Many published studies showed the development of RAPD based marker from the large number of accessions but these plants have good availability such as genus Artemisia [21], Panax species [22], Phyllanthus species [23], Ganoderma lucidum [24], Ipomoea mauritiana [25], Bemisiatabaci (Genn.) [26]. in our study, we used dried root samples for the isolation of the genomic DNA and we successfully isolated the ample amount of genomic DNA by doing some modifications in the CTAB method of Doyle and Doyle (1990)
Summary
In Iridaceae family, Iris is the major genus which has near about 230 species [1]. It provides models for the introgressive hybridization and hybrid fitness studies of the plant [2]. The roots of the plant have many important medicinal properties like it is used to kill parasitic worms, used in hepatic diseases, used as an appetizer, as a diuretic agent, as a detoxifying agent and as an antidote [4,5,6]. This plant is used in combination with other plants to treat various diseases such as liver grievances, dropsy and venereal affections [7]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: International Journal of Pharmacy and Pharmaceutical Sciences
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
