Application of Partial Internal Transcribed Spacer Sequences for the Discrimination of Artemisia capillaris from Other Artemisia Species (original) (raw)
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Biological and Pharmaceutical Bulletin, 2008
Artemisia plants, particularly A. iwayomogi, A. capillaris, A. princeps, and A. argyi, are important medicinal materials that are utilized in traditional Asian medicines. 2) A. iwayomogi and A. capillaris, referred to in Korea as 'Haninjin' and 'Injinho', respectively, 2) have been utilized traditionally for the treatment of diuresis and as anti-inflammatory agents, as is also the case in China and Japan. 2,3) Meanwhile, both A. princeps and A. argyi are referred to as 'Aeyup,' and are employed in the treatment of colic pain, vomiting, and irregular uterine bleeding. 2,4) In traditional Korean medicine markets, a portion of certain specific Artemisia herbs tend to be distributed interchangeably with other Artemisia herbs. 2) For instance, some 'Haninjin' is distributed under the name 'Aeyup.' However, 'Aeyup' may occasionally be substituted with 'Haninjin' or 'Injinho' in medicinal prescriptions. To address this problem, an efficient method for the identification of Artemisia herbs is clearly necessary. Currently, the discrimination of one medicinal herb from another is a process which involves subjective methods, frequently predicated on the morphological features of the plant and/or the observations of a trained individual. However, these methods are not always sufficient for the identification of plants that morphologically resemble each other, particularly when the plants are in a dried and/or sliced state, the common forms of plants when purveyed in an herbal market. Therefore, objective methods, such as those predicated on differences in DNA sequences between organisms, 5) should be developed. In a previous work, 6) we showed that a sequence-characterized amplified region (SCAR) marker 7) based on the nucleotide sequence of a specific random amplified polymorphic DNA (RAPD) product amplified uniquely from 'Aeyup' (both A. princeps and A. argyi) proved to be a reliable DNA marker for the discrimination of 'Aeyup' from other Artemisia herbs. As some Artemisia herbs can be well-distinguished from each other by RAPD, a SCAR marker of 'Haninjin' (A. iwayomogi) developed by the same strategy used for the SCAR markers of 'Aeyup' might facilitate the discrimination of 'Haninjin' from other Artemisia herbs, particularly 'Injinho' and 'Aeyup.' In this work, we developed SCAR markers of Artemisia herbs, including 'Haninjin.' We then applied these SCAR markers to the development of a method that could be applied not only to identify tested Artemisia herbs as 'Haninjin' (A. iwayomogi), but also to determine which Artemisia herbs are being tested concurrently with A. iwayomogi. Using this method, we were able to differentiate 'Haninjin' from other Artemisia herbs with more certainty as compared with a technique predicated on the amplification of a single DNA marker, such as a SCAR marker. Furthermore, we were able to apply this method to a search for Artemisia herbs that are substituted for 'Haninjin' and/or mixed with 'Haninjin'. In service of these research purposes, we developed a multiplex PCR method 8) using a primer set to amplify a SCAR marker of 'Haninjin', along with two other primer sets to amplify the 'Injinho' and 'Aeyup' SCAR markers, in order to amplify each SCAR marker of the Artemisia herbs in a single
Biological and Pharmaceutical Bulletin, 2006
About 500 plants belong to the genus Artemisia. 1,2) Most Artemisia herbs are perennials, growing in the northern hemisphere 3) and are used for various purposes, such as medicine, food, spices, food and ornamentation. The medicinal effects of Artemisia herbs are extremely diverse and include the following: cell protection from peptic ulcers, 4) liver protection, 5,6) anti-malarial 7) and anti-tumor effects. 8) Among Artemisia herbs, A. princeps, A. argyi, A. capillaris, and A. iwayomogi are important medicinal materials in traditional medicine in Asia. In Korea, A. princeps and A. argyi are classified as the same Artemisia herb, called 'Aeyup'. 9) Without discrimination between A. princeps and A. argyi, both plants have been used in traditional Korean medicine for the treatment of colic pain, vomiting and diarrhea, and irregular bleeding from the uterus. 10) On the other hand, A. capillaris and A. iwayomogi are classified as 'Injinho' and 'Haninjin', respectively, and have been used in China and Japan for anti-inflammation and diuresis. 11) For discrimination of medicinal plants, subjective identification methods, based on the plants morphological features, are generally used. With these methods, however, it is very difficult to discriminate between medicinal plants. For instance, young leaves of A. capillaris, harvested particularly in early spring and classified as 'Myun-injin', 9) are not easily distinguished from those of 'Aeyup'. Therefore, a part of 'Injinho' is distributed as 'Aeyup' in the market of traditional Korean medicine. Also, some 'Haninjin' is distributed as 'Aeyup'. Furthermore, instead of 'Aeyup', 'Haninjin' and 'Injinho' are sometimes used for medicinal prescriptions. To solve this problem, an efficient discrimination method, which can be utilized to identify Artemisia herbs, is needed. Various molecular biological techniques, utilizing the different genetic information of organisms, are employed for species discrimination of plants. In particular, random ampli
Research in pharmaceutical sciences
Malaria is currently one of the most important causes of mortality in developing countries. High resistance to available antimalarial drugs has been reported frequently, thus it is crucial to focus on the discovery of new antimalarial drugs. Artemisinin, an effective antimalarial medication, is isolated from various Artemisia species. To identify the Artemisia species producing high quantity of artemisinin, eight species of Artemisia were screened with the genetic sequence characterized amplified region (SCAR) marker for higher quantity of artemisinin. The DNA band corresponding to SCAR marker was cloned into pGEM®-T Easy vector and sequenced. The content of artemisinin in tested species was also measured using high-performance liquid chromatography (HPLC) assay. The primers designed for high-artemisinin SCAR marker could amplify a specific band of approximately 1000 bp which was present in two Artemisia annua and Artemisia absinthium species. These SCAR marker sequences for two sel...
Genomic DNA isolation from Artemisia species grown in cold desert high altitude of India
Artemisia which produces a large number of secondary metabolites is naturally found in cold deserthigh altitude environment of India. Secondary metabolites such as alkaloids, flavonoids, phenols,polysaccharides and terpenes represent a significant barrier to the extraction of pure genomic DNA. Thus, in this study, the DNA extraction protocol to extract pure genomic DNA from different Artemisiaspecies was tailored. The protocol was based on the CTAB method with slight modifications. In thestudy, 1.6 M NaCl, 2% cetyltrimethyl ammonium bromide (CTAB), 3% polyvinylpyrrolidone (PVP) and 0.5% β-mercaptoethanol was used in the extraction buffer. The incubation period was kept for 1 h at65°C with one-tenth of the volume of warm (55°C) 10% CTAB solution during the extraction process.This study described a reliable protocol for extracting good quality and optimum amount of DNA from Artemisia species suitable for PCR analysis.
Journal of Essential Oil Bearing Plants, 2019
The present study aims to investigate the chromatographic fingerprinting of the essential oils of Artemisia vulgaris L. growing wild in Kumaon and Garhwal region of Uttarakhand, India. The aerial parts were harvested in full flowering time from twenty two locations at the altitudes between 627-2793 m. The essential oils from aerial parts were isolated by hydro-distillation through a Clevenger apparatus and analysed by GC and GC/MS for the identification of their chemical composition. Chromatographic analysis has shown that all the essential oils of A. vulgaris were dominated by varied composition and chrysanthenone (0.1-26.6 %), vulgarole (0.1-20.6 %), artemisia ketone (0.01-19.8 %), α-thujone (0.01-19.0 %), 1,8-cineole (1.6-13.5 %), β-thujone (0.2-13.2 %), caryophyllene oxide (1.4-11.2 %) and camphor (0.9-11.1 %). Hierarchical cluster and principal component analysis (PCA) were applied to study the chemical variability among populations which showed three major groups. Group-1 having A17, A38, A42, A46 and A56 populations, was possessed vulgarole (7.6-20.6 %) in higher amount followed by γ-eudesmol (4.9-9.2 %), camphor (1.4-10.0 %), 1,8-cineole (1.6-8.9 %) and camphene (1.3-7.5 %). In group 2 (A52, A60), artemisia ketone was found in major portions (18.9-19.8 %) followed by 1,8-cineole (2.6-13.5 %) and artemisia alcohol (1.8-8.6 %), while group 3 (
2020
Artemisia is a significant genus in family Asteraceae and has large number of taxa having economic and ecological importance. The aim of the study was to analyze highly medicinal products as well, antioxidant activity and genetic variability in three species of Artemisia from various sites of district Poonch. For the evaluation of phytochemicals and antioxidants, different volumes of extracts and solvents were used from leaves. The species were analyzed for genetic diversity within and between various Artemisia species. For molecular investigation, random amplification of polymorphic DNA (RAPD) fragment was used to determine genetic variations between and within species. From of ten primers only six primers gave amplified products. By using all six primers, total 331 bands were amplified from which 30 were monomorphic, 301 were polymorphic and percentage of polymorphism was 91% in Artemisia japonica, 321 bands were amplified in Artemisia roxberghiana from which 21 were monomorphic, 3...
Applying high-resolution melting (HRM) technology to identify five commonly used Artemisia species
Scientific reports, 2016
Many members of the genus Artemisia are important for medicinal purposes with multiple pharmacological properties. Often, these herbal plants sold on the markets are in processed forms so it is difficult to authenticate. Routine testing and identification of these herbal materials should be performed to ensure that the raw materials used in pharmaceutical products are suitable for their intended use. In this study, five commonly used Artemisia species included Artemisia argyi, Artemisia annua, Artemisia lavandulaefolia, Artemisia indica, and Artemisia atrovirens were analyzed using high resolution melting (HRM) analysis based on the internal transcribed spacer 2 (ITS2) sequences. The melting profiles of the ITS2 amplicons of the five closely related herbal species are clearly separated so that they can be differentiated by HRM method. The method was further applied to authenticate commercial products in powdered. HRM curves of all the commercial samples tested are similar to the bot...