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The present study utilizes polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using partial plastid rbcL and mitochondrial trnC–trnP gene sequences to distinguish the six representative Pyropia species produced via mariculture in Korea. The rbcL, trnC, and trnP sequences of 15 Pyropia species from the NCBI database were aligned to determine specific restriction enzyme sites of the six Pyropia species. To confirm the presence of restriction sites of eight enzymes, PCR amplicons were digested as follows: a 556 bp fragment within the rbcL region of chloroplast DNA was confirmed in P. yezoensis using BglI, whereas Tth111I, AvaII, BsrI, and BsaAI enzymes produced fragments of 664, 271, 600, and 510 bp, respectively, from the rps11–trnG region of mitochondrial DNA in P. seriata, P. dentata, P. suborbiculata, and P. haitanensis. In the case of P. pseudolinearis, HindIII, SacII, and SphI enzymes each had two cleavage sites, at positions 174 and 825, 788 and 211, and 397 and 602 bp, respectively. All six species were successfully distinguished using these eight restriction enzymes. Therefore, we propose that PCR-RFLP analysis is an efficient tool for the potential use of distinguishing between the six Pyropia species cultivated via mariculture in Korea.

This study investigated the antioxidant, antiaging, and antibacterial properties of Gracilaria verrucosa (GV) based on 95% methanol (GVM), ethanol (GVE), and hot water (GVW) extractions. Antioxidant activity assays revealed the total polyphenol and flavonoid contents were highest in GVM and GVE. The 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-Azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) activities were highest in GVE and GVM. Furthermore, GVE exhibited the highest ferric-reducing antioxidant power (FRAP) value. In comparison, superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities were highest in GVM. Collectively, GVE and GVM had stronger antioxidant activities than GVW. Additionally, collagenase, elastase, and tyrosinase inhibition assays showed that GVM exhibited the strongest anti-wrinkle and skin-whitening activities. Liquid chromatography–tandem mass spectrometer (LC–MS/MS) revealed that GVW had the highest 4-hydroxy benzoic acid content, whereas GVE had the highest naringenin and naringin contents. Additionally, GVE exhibited the strongest antimicrobial activity against six foodborne bacteria, with minimum inhibitory and bactericidal concentrations of 0.06–0.3 and 0.1–0.5 μg/μL. Correlation analysis of the GV extracts indicated a strong positive relationship between TPC and ABTS, SOD, and CAT activities (r = 0.760–0.982, p = 0–0.018). Overall, GVE and GVM can be applied to the development of functional agents across diverse industries.

The tuber of Cynanchum wilfordii (Baekshuoh Radix in Korean) is an important medicinal herb in Korea and China; however, it is difficult to differentiate C. wilfordii from a related medicinal herb, C. auriculatum (Baishouwu Radix in Chinese). We sought to develop a molecular method that could be used to distinguish between the tubers of C. wilfordii and C. auriculatum. We aligned the chloroplast genome sequences (available in the NCBI database) of the two species and identified three species-specific insertion and deletion (InDel) sites in the trnQ-psbK, rps2-rpoC2, and psaJ-rpl33 intergenic spacer (IGS) regions. To confirm the presence of these three InDels and validate their use as markers, we designed three primer pairs to amplify the trnQ-psbK, rps2-rpoC2, and psaJ-rpl33 IGS regions. Polymerase chain reaction (PCR) amplification of the trnQ-psbK IGS region yielded a 249 bp fragment for C. wilfordii, and 419 bp fragment for C. auriculatum, whereas the rps2-rpoC2 IGS primers produced a 629 bp fragment from C. wilfordii and a 282 bp fragment from C. auriculatum. In the psaJ-rpl33 IGS region, allele fragments of 342 and 360 bp in length were amplified from C. wilfordii, whereas 249 and 250 bp fragment were amplified from C. auriculatum. We propose these three InDel markers as a valuable, simple, and efficient tool for identifying these medicinal herbs and will thus reduce adulteration of these herbal materials in commercial markets.

We previously established that polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using partial plastid rbcL and mitochondrial trnC–trnP gene sequences can be used to distinguish the six representative Pyropia species produced via mariculture in Korea. In this study, we develop progressive InDel markers by comparing seven complete Pyropia chloroplast genomes obtained from The National Center of Biotechnology Informnation (NCBI) GenBank. Comparative analyses of nucleotide diversity among the genomes revealed seven hypervariable sites (cemA, rps13, trnM-argB, petD-petB, trnR-trnQ, ccs1-orf24, and ycf12-ftrB) among 637 sliding windows with nucleotide diversity > 0.025 (Pi). These sites included two genes and five gene-intergenic regions, three of which (cemA, trnM-argB, trnR-trnQ) showed complete amplification for all six test species. Finally, trnM-argB, an InDel-variable locus with high discriminatory power, was selected as a DNA barcode candidate. These results suggest that the obtained trnM-argB region can be used for the effective exploration of the variation present in six Korean Pyropia and for further evolutionary, phylogenetic, barcoding and genetic engineering studies of Pyropia species.

The leaves of Perilla frutescens var. acuta (PFA) are commonly used as a traditional medicine in Korea, Japan, and China. We previously showed that PFA attenuates eye fatigue by improving visual accommodation through a clinical study. However, detailed mechanisms and chemical compounds have not been studied. In this study, we analyzed the active compounds in an aqueous extract of PFA involved in ciliary muscle relaxation in vitro and in vivo. NMR and MS analyses showed that the PFA extract contained mainly luteolin-7-O-diglucuronide and apigenin-7-O-diglucuronide. The composition after freeze-drying and spray-drying was similar. Freeze-dried PFA (50 µg/mL, 100 µg/mL, and 200 µg/mL) increased nitric oxide and cGMP levels in ciliary muscle cells isolated from the eyes of rats. [Ca2+]i decreased in a dose-dependent manner. Furthermore, Sprague-Dawley rats treated with freeze-dried PFA (200 mg/kg, orally) showed significantly increased cGMP levels compared with the control group and irradiated with white light. Our results suggest that PFA extract has the potential to reduce eye fatigue by relaxing ciliary muscles.

Members of the Vaccinium genus (Ericaceae family) have excellent medicinal and nutritional properties. In this study, we sequenced the complete chloroplast (cp) genomes of three species, Vaccinium bracteatum Thunb., V. vitis-idaea L., and V. uliginosum L., to investigate their phylogenetic relationships within the Ericaceae. The chloroplast genomes of these species are 174,404 bp, 173,967 bp, and 173,356 bp, respectively. These genomes have a typical quadripartite structure, with an LSC region (106,565 bp, 106,013 bp, and 105,856 bp) and an SSC region (2979 bp, 3518 bp, and 3146 bp) separated by a pair of IRs (32,430 bp, 32,218 bp, and 32,177 bp, respectively). These cp genomes are composed of 139 genes, including 93 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. Maximum likelihood phylogenetic analysis indicated that V. bracteatum is most closely related to V. oldhamii. Our findings should facilitate further evolutionary and genetic studies.

The complete chloroplast (cp) genome of Stauntonia hexaphylla, a monotypic genus native to Korea, was determined. The whole cp genome is 158,390 bp in size, containing a large single-copy (LSC) region of 87,115 bp and a small single-copy region (SSC) of 18,928 bp, separated by a pair of inverted repeats (IRs) of 26,174 bp. The cp genome encodes 117 genes, including 79 protein-coding, 38 tRNA-coding, and 8 rRNA-coding genes. The overall GC content is 37.8%. A phylogenetic analysis demonstrated a close relationship between S. hexaphylla and S. obovatifoliola subsp. urophylla. The cp genome will provide new insight into the evolution of Lardizabalaceae.

Zanthoxylum schinifolium and Zanthoxylum piperitum are the sources of the well-known traditional Korean herbal medicines “sancho” (prickly ash) and “chopi” (Korean pepper), respectively. Sancho and chopi are often indiscriminately mixed due to the similar appearance of the herbal materials when used as spices and herbal medicines. Moreover, commercial sancho and chopi products often contain adulterants, which is insufficient to ensure food efficacy and safety. In this study, we developed hypervariable insertion/deletion (InDel) markers to distinguish between sancho and chopi products by comparing the complete chloroplast genome sequences of four Zanthoxylum species deposited in the National Center for Biotechnology Information (NCBI) GenBank. Comparative analyses of the nucleotide diversity (Pi) of these Zanthoxylum genomes revealed four hypervariable divergent sites (trnH-psbA, psbZ-trnG, trnfM-rps14, and trnF-ndhK) with Pi > 0.025 among 520 windows. Of these four regions, including two genic and two intergenic regions, only psbZ-trnG yielded accurate PCR amplification results between commercial sancho and chopi products from the Korean herbal medicine market. We therefore selected psbZ-trnG, an InDel-variable locus with high discriminatory powers, as a candidate DNA barcode locus. This InDel marker could be used as a valuable, simple, and efficient tool for identifying these medicinal herbs, thereby increasing the safety of these spices and herbal materials in the food market.

Pyropia dentata is an economically valuable species in seaweed aquaculture in the southwest coastal regions of Korea. Herein, we determined the complete chloroplast (cp) genome sequence of P. dentata using PacBio resequencing data. The whole cp genome is 192,266 bp in size, encodes 148 genes (including 92 protein-coding, 44 tRNA-coding, and 12 rRNA-coding genes), and lacks inverted repeat (IR) regions. A maximum likelihood phylogenetic analysis demonstrated that P. dentata was most closely related to P. haitanensis. The cp genome will provide a useful reference for further studies of P. dentata.

Elaeagnus is a genus which consists about 70 species of flowering plants in the family Elaeagnaceae, and its edible fruit is a natural product used as food and in traditional medicine. In this study, we sequenced the complete chloroplast (cp) genome of four species, namely Elaeagnus umbellate Thunb., E. multiflora Thunb., E. macrophylla Thunb., and E. glabra Thunb., to study their phylogenetic relationships within the Elaeagnaceae. Total lengths of the chloroplast genome were 152,261 bp, 152,267 bp, 152,224 bp, and 152,227 bp, respectively. The four genomes had representative quadripartite structures, with an LSC region (82,207 bp, 82,191 bp, 82,136 bp, and 82,139 bp) and an SSC region (18,262 bp, 18,282 bp,and 18,278 bp for both species) separated by a pair of IRs (25,896 bp, 25,897 bp, and 25,905 bp for the latter two species), respectively. Moreover, they were composed of 136-137 genes, including 88 protein-coding genes, 40-41 tRNA genes, and 8 rRNA genes. A maximum likelihood phylogenetic analysis indicated that E. umbellata was most closely related to E. multiflora, whereas E. macrophylla was close to E. glabra.