Development of molecular markers for authentication of the medicinal plant species Patrinia by random amplified polymorphic DNA (RAPD) analysis and multiplex-PCR

Abstract

The authentication of components from plant species is vital for the quality control and standardization of herbal medicines. Due to morphological similarities when dried, it is very difficult to distinguish authentic medicinal plants Patrinia villosa and P. scabiosifolia from adulterants of P. rupestris and P. saniculifolia in the herbal medicine Patriniae Radix. To establish a reliable authentication tool distinguishing authentic Patriniae Radix species from adulterants, we used random amplified polymorphic DNA (RAPD) genomic profiling and obtained several species-specific DNA fragments for each of the four Patrinia species. Based on the sequences of these amplicons, we developed sequence characterized amplified region (SCAR) markers as stable molecular authentication tools. SCAR markers to identify the authentic Patriniae Radix species, P. villosa and P. scabiosifolia, were developed. To accurately identify adulterants, we also developed SCAR markers able to identify the inauthentic P. rupestris and P. saniculifolia species. Furthermore, we established multiplex-PCR SCAR assays to simultaneously distinguish all four species using combinations of species-specific SCAR marker primer sets. These efforts resulted in reliable DNA-based molecular markers to identify authentic Patriniae Radix species from their adulterants and a rapid authentication assay for the efficient identification of Patrinia species. These markers will facilitate the standardization of Patriniae Radix preparations and therefore prevent the distribution of adulterants.

Differentiating Authentic Adenophorae Radix from Its Adulterants in Commercially-Processed Samples Using Multiplexed ITS Sequence-Based SCAR Markers

“…Judging from our laboratory experience, these additional costs amount to at least $300-$400 USD for the analysis of 10 herbal medicine samples. Therefore, the SCAR method is much more convenient and less expensive for the authentication of herbal medicines and for the identification of the botanical origins of herbal medicines than the DNA barcoding method [22,24,26]. In spite of these disadvantages, DNA barcoding is mightily important for plant species identification with high resolution and for obtaining primary sequence information in species-specific marker development.…”

“…Therefore, the SCAR method is simple to carry out, its results are simple to interpret, and it produces fewer errors in PCR amplification and sequencing [21]. Furthermore, the efficiency and ability of this assay improved with the combination of individual SCAR markers and multiplex PCR, namely, multiplexed SCAR marker assays [22,25,26]. These have been abundantly and successfully applied to authenticate medicinal plant species and commercially-processed herbal medicines [22].…”

Espécies de Ralstonia no Brasil : caracterização fenotípica, molecular, novas fontes de resistência em tomateiro e patogenicidade em cafeeiro

“…Marcadores moleculares são uma ferramenta importante para a pesquisa genética devido a sua capacidade de diferenciar e estudar variações fenotípicas e genotípicas (Moon et al, 2016). Os sistemas de marcadores moleculares (ex.…”

“…A estratégia de marcadores SCAR (sequence characterized amplified region) emprega, como informação inicial para geração de primers específicos, a sequencia de diversos marcadores usados para fingerprinting (Holmberg et al, 2009), tais como RAPD (Moon et al, 2016) e BOX-PCR (Rossi et al, 2014). Amplicons são purificados e sequenciados, suas sequencias são então usadas para desenho de primers específicos.…”

Cultivation and Characteristics of Licorice F1 (Glycyrrhiza glabra x G. uralensis) Lines

“…Medicinal plants have become highly valued as potential herbal medicines in many countries. Thus, various molecular markers are development such as RAPD, ITS/NTS, SCAR (Kim et al, 2015;Moon et al, 2016). They have been used for authenticate and distinguish plants from their relatives.…”