Development of real-time PCR based molecular markers for two medicinal herb Artemisia species A. capillaris and A. iwayomogi.

Artemisia capillaris and Artemisia iwayomogi are well-known herbal medicines which are used as hepatotherapeutic drugs. These two herbal species can be confused with each other, owing to their morphological similarity and similar Korean common names of "Injinho" and "Haninjin," respectively. Molecular markers to distinguish between the two plants were developed. Six primer sets were designed and verified, and their efficiencies were found to range from 90.28 to 98.29%. The developed primer sets had significant correlation coefficient values between the cycle threshold values and the logarithm of DNA concentration for their target species (R2 > 0.98), with slopes ranged from - 3.3637 to - 3.5793. The specificity of the quantitative polymerase chain reaction (qPCR) was confirmed with 14 other species. Additionally, 16 commercial medicinal herbs and 40 blind samples were tested to evaluate their reliability. Collectively, the findings indicate that developed qPCR-based target-specific primer sets have potential applicability toward protection of consumers' rights. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01166-0.

Complete organellar genomes of six Sargassum species and development of species-specific markers.

Sargassum is one of the most important brown algal genera that can be used as food and raw material for medicinal purpose, and has various beneficial effects. As the classification of Sargassum species is currently based on their morphological characteristics, organellar genome sequences of Sargassum would provide important information for accurate identification of species and developing species-specific markers. We sequenced the complete organellar genomes of six Sargassum species, including the first complete chloroplast genome sequences of S. fulvellum, S. serratifolium, S. macrocarpum, and S. siliquastrum, and the first complete mitochondrial genome sequences of S. fulvellum, S. serratifolium, and S. macrocarpum. The chloroplast genomes of the 6 Sargassum species contained 139 protein-coding genes (PCGs), and the mitochondrial genomes possessed 37 PCGs. A comparative study was performed between the newly sequenced organellar genomes and 44 other species belonging to class Phaeophyceae. Phylogenetic relationships using PCGs shared by Phaeophyceae species were constructed with IQ-TREE 2 using the maximum likelihood method. In addition, we developed real-time PCR markers based on SNPs to distinguish the 6 Sargassum species. Our results provide useful information for establishing phylogenetic relationships between brown algae.

Development and application of DNA markers to detect adulteration with Scopolia japonica in the medicinal herb Atractylodes lancea.

Atractylodes lancea rhizomes are commonly consumed in east Asia as traditional medical herbs. However, in Korea, because of their morphological similarity, A. lancea rhizomes can be contaminated with those of Scopolia japonica imported from China. To detect adulteration with S. japonica in the complex products of A. lancea, we developed two PCR-based DNA markers, multiplex PCR and quantitative real-time PCR. The sensitivity of the multiplex PCR primer combinations and real-time PCR was confirmed with a series of DNA concentrations (0.01-10 ng/µL). The specificity of the developed PCR assays was confirmed with 14 other species. In addition, 14 commercial A. lancea medicinal herbs and 20 blind samples were tested with the developed PCR assays to demonstrate the reliability. Taken together, the developed multiplex and real-time PCR-based target-specific primer sets may be useful for detecting the target species and have the potential to contribute to food safety and consumer health care. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-01008-5.