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ABSTRACT: One of the medicinal materials produced by plants belonging to the genus Scrophularia, ‘Scrophularia Radix’ (SR), has been prescribed for many centuries to treat diseases such as inflammation, abscesses of carbuncles
and constipation. In China, the dried root of S. ningoensis Hemsley is the source of SR. In contrast, the root of S. buergeriana is generally prescribed as SR in Korea. Studies conducted to identify the bioactive compounds in these two Scrophularia plants have revealed marked differences in the contents and concentration of the compounds they contain. However, S. ningpoensis has been indiscriminately prescribed in Korea along with S. buergeriana as SR. Furthermore, S. koraiensis has long been used in lieu of S. buergeriana as SR in Korea. Therefore, a standard or method to reliably distinguish these three species of Scrophularia is needed. Recently, we found that the differences in the nucleotide sequences of the internal transcribed spacer (ITS) of
Scrophularia plants could be applied to develop DNA markers to discriminate each plant. In this study, ITS sequences of 22 samples including
three types of Scrophularia plants were amplified, determined and analyzed. Based on the results of these analyses, we designed the following primer
sets: Ni F (5′-TTAACCATATAGGGGCCTCG-3′) / Ni R (5′-C CCCTCTCTGTATCCCAA-3′) to amplify a 379 bp DNA marker for the identification
of S. ningpoensis; Bu F (5′-TTAACC ATATCGGGGCCAAG-3′) / Bu R (5′-ATCACGACAGCAC GCGA-3′) to amplify a 491 bp DNA marker for S. buergeriana; and Ko F (5′-ATAACCATATCGGGGCCTC-3′) / Ko R (5′-TCAAGAAACGCACTATCCC-3′) to amplify a 167 bp DNA marker of S. koraiensis. Using these primer sets, we were able to efficiently identify Scrophularia plants sold as SR in the herbal market in dried and sliced states after processing as medicinal materials.
KeywordsScrophularia Radix-
S. buergeriana
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S. ningpoensis
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S. koraiensis
-DNA maker-ITS, internal transcribed spacer
Genes & genomics 04/2012; 32(2):181-189. · 0.44 Impact Factor
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ABSTRACT: This study describes a method for discriminating Rangifer antlers from true Cervus antlers using agarose gel electrophoresis, capillary electrophoresis, quantitative real-time PCR, and allelic discrimination. Specific primers labeled with fluorescent tags were designed to amplify fragments from the mitochondrial D-loop genes for various Cervus subspecies and Rangifer tarandus differentially. A 466-bp fragment that was observed for both Cervus and Rangifer antlers served as a positive control, while a 270-bp fragment was specifically amplified only from Rangifer antlers. Allelic discrimination was used to differentiate between Cervus and Rangifer antlers, based on the amplification of specific alleles for both types of antlers. These PCR-based assays can be used for forensic and quantitative analyses of Cervus and Rangifer antlers in a single step, without having to obtain any sequence information. In addition, multiple PCR-based assays are more accurate and reproducible than a single assay for species-specific analysis and are especially useful in this study for the identification of original Cervus deer products from fraudulent Rangifer antlers.
Journal of Animal Science 01/2012; 90(7):2075-83. · 2.10 Impact Factor
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ABSTRACT: Some plants classified in the genus Artemisia are used for medicinal purposes. In particular, A. iwayomogi, which is referred to as 'Haninjin,' is used as an important medicinal material in traditional Korean medicine. However, A. capillaris, and both A. argyi and A. princeps, referred to as 'Injinho' and 'Aeyup,' respectively, are used for purposes other than those for which 'Haninjin' is utilized. However, it is occasionally difficult to differentiate 'Haninjin' from 'Injinho' and/or 'Aeyup' on the basis of their morphological features, particularly when in the dried and/or sliced form. Therefore, the development of a reliable method by which to discriminate 'Haninjin' from other Artemisia herbs, especially 'Injinho' and 'Aeyup,' is clearly necessary. We recently determined that the RAPD (random amplified polymorphic DNA) technique can be used to discriminate efficiently between some Artemisia herbs. In particular, when applied to RAPD, the non-specific UBC primer 391 (5'-GCG AAC CTC G-3') was demonstrated to amplify PCR products specific to A. iwayomogi. Based on the nucleotide sequences of the PCR product, we designed a 2F1 (5'-ACC TCG GAC CTA AAT ACA-3')/ 2F3 (5'-TTA TGA TTC ATG TTC AAT TC-3') primer set to amplify a SCAR (sequence-characterized amplified region) marker of A. iwayomogi. Employing this primer set, along with two other primer sets amplifying SCAR markers of 'Aeyup' (A. argyi and A. princeps) and both 'Injinho' (A. capillaris) and A. japonica, which are classified into the same subgroup in a phenogram constructed from RAPD analysis, we developed a multiplex PCR method by which A. iwayomogi could be discriminated with certainty from other Artemisia herbs. Via this method, we determined not only whether the tested Artemisia herb was A. iwayomogi, but also which Artemisia herbs were tested concurrently with A. iwayomogi.
Biological & Pharmaceutical Bulletin 05/2008; 31(4):685-90. · 1.66 Impact Factor
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ABSTRACT: Some Artemisia herbs are used for medicinal purposes. In particular, A. princeps and A. argyi are classified as 'Aeyup' and are used as important medicinal material in traditional Korean medicine. On the other hand, A. capillaris and A. iwayomogi, which are classified as 'Injinho' and 'Haninjin', respectively, are used for other purposes distinct from those of 'Aeyup'. However, sometimes 'Aeyup' is not clearly discriminated from 'Injinho' and/or 'Haninjin'. Furthermore, Artemisia capillaris and/or A. iwayomogi have been used in place of A. princeps and A. argyi. In this study, we developed an efficient method to discriminate A. argyi and A. princeps from other Artemisia plants. The RAPD (random amplified polymorphic DNA) method efficiently discriminated various Artemisia herbs. In particular, non-specific primer 329 (5'-GCG AAC CTC C-3'), which shows polymorphism among Artemisia herbs, amplified 838 bp products, which are specific to A. princeps and A. argyi only. Based on nucleotide sequence of the primer 329 product, we designed a Fb (5'-CAT CAA CCA TGG CTT ATC CT-3') and R7 (5'-GCG AAC CTC CCC ATT CCA-3') primer-set to amplify a 254 bp sized SCAR (sequence characterized amplified regions) marker, through which A. princeps and A. argyi can be efficiently discriminated from other Artemisia herbs, particularly, A. capillaris and A. iwayomogi.
Biological & Pharmaceutical Bulletin 05/2006; 29(4):629-33. · 1.66 Impact Factor
