Development of SCAR Markers for the Discrimination of Three Species of Medicinal Plants, Angelica decursiva (Peucedanum decursivum), Peucedanum praeruptorum and Anthricus sylvestris, Based on the Internal Transcribed Spacer (ITS) Sequence and Random Amplified Polymorphic DNA (RAPD)

Abstract

Angelicae decursivae radix ('Jeonho' in Korean) is prescribed as the root of Angelica decursiva (= Peucedanum decursivum) and Peucedanum praeruptorum in Korean pharmacopoeia. However, Anthricus sylvestris has been usually distributed on the market because it is identical to the Korean plant name 'Jeonho'. Furthermore, due to the morphological similarity of the aerial parts and herbal medicines, the correct identification of these roots is difficult. Therefore, to develop a reliable method for discriminating among A. decursiva (= P. decursivum), P. praeruptorum and A. sylvestris, we applied the tools of molecular genetics, such as the analysis of ribosomal DNA internal transcribed spacer (rDNA-ITS) and random amplified polymorphic DNA (RAPD). In the comparison of rDNA-ITS sequences, we found a specific primer region for the identification of A. sylvestris among three varieties of the herb that produced a 273 bp strand of DNA specific to A. sylvestris. As the result of RAPD analysis, we developed one sequence characterized amplified region (SCAR) marker for A. decursiva and P. praeruptorum that amplified a 363 bp DNA fragment specific to both A. decursiva and P. praeruptorum and two markers for P. praeruptorum that amplified 145 bp and 305 bp DNA fragments specific to P. praeruptorum. Furthermore, we established the SCAR markers for the simultaneous discrimination of the three species by applying a multiplex-polymerase chain reaction (PCR) with a combination of primers. This method of discrimination would be useful in preventing the distribution of adulterates because it can identify each herb and distinguish it from inauthentic substitutions.

SCAR markers for discriminating species of two genera of medicinal plants, Liriope and Ophiopogon

“…PCR-based identification is particularly useful when botanical identification based on morphology is difficult, such as when samples are incomplete, damaged, and/or dried (Ray and Roy, 2009). Choo et al (2009) used multiplex-PCR, which is rapid and highly specific for DNA of closely related species, to successfully amplify specific markers from mixed DNA samples. Such efficient, precise and sensitive techniques are required for rapid identification of variants at the interand/or intraspecies level (Weder, 2002).…”

“…Because it is rapid, simple and relatively inexpensive, particularly to analyze numerous samples, the SCAR method has been used to identify a wide range of medicinal plant species, including Panax ginseng (Wang et al, 2001), Artenisia princeps and A. argyi (Lee et al, 2006), Phyllanthus emblica (Dnyaneshwar et al, 2006), Anthricus sylvestris (Choo et al, 2009), Bambusa balcooa (Das et al, 2005) and Atractylodes (Chen et al, 2001).…”

Identification 4 kinds of Muxiang using Multiplex PCR

“…. ITS 부위의 염기서열은 진화에 따라 빠르게 변화하기 때문에, ITS 염기서열 정보는 특히 식물의 속 (genus) 이하 단위에서의 계통학적 유연관계 연구 [21][22][23] , 유전 적 다양성 평가 24) , 종 내 변이 연구 25) , DNA barcoding 26) 을 위해 널리 사용되고 있다. 이러한 ITS 염기서열은 약용식물의 감별을 위해서도 효율적으로 사용되고 있다 27) .…”

Molecular authentication of the medicinal herb Ruta graveolens (Rutaceae) and an adulterant using nuclear and chloroplast DNA markers

“…Therefore, nrDNA-ITS region is believed to be a preferred choice for molecular typing of plant species. Moreover, in recent years, ITS sequence of nrDNA has been employed as genetic markers in various plant species (Zhang et al, 2007;Choo et al, 2009;Gulbitti-Onarici et al, 2009). These nrDNA-ITS fragments amplified from the genomic DNA samples of R. graveolens and E. dracunculoides were sequenced and sequence differences were observed through the Clustal-X software.…”

“…Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA) have been extensively used for phylogenetic study among plant species (Choo et al, 2009;Gulbitti-Onarici et al, 2009), molecular discrimination of herbal materials (Zhang et al, 2007) and DNA barcoding (Zuo et al, 2011). The nrDNA-ITS region has frequent insertions/ deletions, which can be phylogenetically informative (Baldwin et al, 1995).…”