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Background: The genus Prunus (Family Rosaceae) comprises over 400 plant species and exhibits vast biodiversity worldwide. Due to its wide distribution, its taxonomic classification is important. Anatomical characters are conserved and stable and thus can be used as an important tool in plant taxonomic characterization. Thus, this study aimed at examining and documenting P. africana leaf, stem, and seed anatomy using micrographs and photographs for possible use in identification, quality control, and phylogenetic studies of the species.Methods: P. africana leaves, stems, and seeds were fixed, dehydrated in ascending ethanol series (50–100 %), embedded in Technovit resin, and sectioned using a microtome for mounting histological slides for anatomical observation under a microscope and subsequent description.Results: The anatomical sections of a young stem revealed a cortex consisting of isodiametric parenchyma cells, druse crystals, primary vascular bundles, and pith. The mature stem bark consisted majorly of rhytidome with periderm densely arranged in multiple layers, a cluster of stone cells, and sclerenchyma. The sections of the leaf were hypostomatic with stomata size ranging between 18.90– (22.34)–26.90 × 15.41– (18.40)–21.22 μm. The leaf sections showed the presence of characteristic druse crystals, vascular bundles, and mesophyll layers. The pericarp showed the presence of epicarp, mesocarp, and endocarp with a thickness of approximately 350–400, 300–350, and 30–50 μm, respectively and a seed testa with a thickness of approximately 50–60 μm. Conclusion: The characteristic morphological and anatomical features observed in P. africana leaves, stems, and seeds in this study could provide useful data in taxonomical identification of this species.

The genus Fritillaria is an important herbaceous medicinal plant. The dried bulbs of many Fritillaria species are used as Fritillariae Bulbus in traditional oriental medicines. However, these herbal medicines frequently contain mixtures of different types of Fritillariae Bulbus and other Fritillaria species with little or no medicinal value. The accurate identification of different types of Fritillariae Bulbus is very difficult due to the morphological similarities between the plants and dried bulbs; therefore, it is necessary to accurately identify these herbal medicines and other Fritillaria species based on their genetic characteristics. To find molecular markers for Fritillaria species and elucidate the genomic characteristics of F. thunbergii, we sequenced the complete chloroplast (cp) genome of F. thunbergii and performed a comparative analysis with related plant taxa. The complete cp genome of F. thunbergii is 152,155 bp with a quadripartite structure, which is similar to other plants, and was highly similar to previously reported Fritillaria cp genomes in regard to gene content, order, orientation, and GC contents. Comparative analysis of the cp genomes identified two genes, matK and rps16, that contained sufficient markers to distinguish between two herbal medicines (Fritillariae Thunbergii and Fritillariae Hupehensis Bulbus) and five Fritillaria species. These results provide useful information for identification of Fritillaria species and quality control of Fritillariae Bulbus.

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.

To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared (FT-IR) spectroscopy can be used to discriminate and compare metabolic equivalence, standard medicinal parts from four medicinal plants (Cynanchum wilfordii Hemsley, Atractylodes japonica Koidz, Polygonum multiflorum Thunberg and Astragalus membranaceus Bunge) and their in vitro-produced adventitious roots were analyzed by FT-IR spectroscopy. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from the FT-IR spectral data showed that the whole metabolic pattern from Cynanchum wilfordii was highly similar to Astragalus membranaceus. However, Atractylodes japonica and Polygonum multiflorum showed significantly different metabolic patterns. Furthermore, adventitious roots from Cynanchum wilfordii and Astragalus membranaceus also showed similar metabolic patterns compared to their standard medicinal parts. These results clearly show that mass proliferation of adventitious roots may be applied to aquire novel supply of standard medicinal parts from medicinal plants. However, the whole metabolic pattern from adventitious roots of Atractylodes japonica and Polygonum multiflorum were not similar to their standard medicinal parts. Furthermore, FT-IR spectroscopy combined with multivariate analyses established in this study may be applied as an alternative tool to discriminate the whole metabolic equivalence from several standard medicinal parts. Thus, we suggest that these metabolic discrimination systems may be applied for metabolic standardization of herbal medicinal resources.

Akebiae Caulis, ‘Mutōng’ in Chinese, refers to the stem of Akebia quinata in the Korean modern pharmacopoeia. The stems of Aristolochia manshuriensis and Clematis armandii have also been traditionally used for clinical purposes, and are often marketed as ‘Mutōng’ under the names ‘Guān-Mutōng and Chuān-Mutōng’, respectively. Furthermore, due to the morphological similarity of the herbal medicines, their correct identification by traditional subjective methods is difficult. Therefore, to develop a reliable method for discriminating these three herbal medicines, we applied the tools of molecular genetics, including random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR) marker development using samples of Akebia quinata, Akebia trifoliata, Aristolochia manshuriensis, and Clematis armandii. By comparative analysis of RAPD polymorphisms, we identified several potential PCR products that can be used to distinguish the herbal materials of each species. Using these RAPD products, we developed unique sequence characterized amplified region (SCAR) markers for determination of the species of plant materials; the products were 590 bp for A. quinata, 300 bp for A. trifoliata, 373 bp for A. manshuriensis, and 432 bp for C. armandii. Furthermore, we optimized SCAR markers for simultaneous discrimination of the four species by multiplex-PCR. These markers allow efficient identification of closely related herbal medicines originating from Akebia, Aristolochia, and Clematis species, and have potential uses in preventing the distribution of adulterants, as they can be used to identify each species and distinguish them from inauthentic substitutes.