Increased public awareness of nutritional and health issues has resulted in the increasing consumption of food and herbal products made from the root of Pueraria montana var. lobata (Willd.) Maesen & S. M. Almeida ex Sanjappa & Predeep (kudzu vine) and P. montana var. thomsonii (Benth.) M. R. Almeida. The famous herbal medicine Yufeng Ningxin, which is used to treat cardiovascular diseases, can be legally produced only using P. montana var. lobata. However, precise identification at the subspecies level is usually challenging when these products’ ingredients lose their morphological characteristics after deep processing. Here, six herbarium specimens, 21 expert-identified original plant samples, 30 raw material samples, 10 food products and 12 herbal products were collected to test the subspecies-level authentication abilities of ITS2 sequences. The results showed that ITS2 sequences can distinguish P. montana var. lobata from P. montana var. thomsonii with stable single nucleotide polymorphism (SNP) sites. A total of 93.3% of raw material samples were consistent with the markings on their labels, but only 50% of Gegen Powder samples were made from P. montana var. lobata. High-quality ITS2 sequences were successfully obtained from nine of the 12 herbal products using Sanger sequencing. Substitution and fungal contamination were detected in 3 herbal products by further DNA metabarcoding, even though thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) tests verified that the products met existing quality standards. This study demonstrated that DNA barcoding is a powerful tool for the identification of P. montana var. lobata and P. montana var. thomsonii at the subspecies level, and we conclude that DNA barcodes can be broadly applied to trace the raw materials of food and herbal products.

Bupleuri Radix is the dry root of certain species of the genus Bupleurum and is commonly used in traditional Chinese medicine. The increasing global demand for Bupleuri Radix cannot be fulfilled with wild populations only. Therefore, cultivated Bupleurum is now the main commercial source of this medicinal product. Different species of Bupleurum show different medicinal properties and clinical effects, making reliable authentication and assignment of correct botanical origin for medicinal species critical. However, accurate identification of the cultivated Bupleurum species is difficult due to dramatic morphological variations resulting from cultivation. In this study, we sampled 56 cultivated Bupleurum populations of six different morphotypes (Types A-F) from the main production areas of China, and 10 wild populations of four species were used as reference materials. Conventional DNA barcoding was conducted to identify cultivated Bupleurum species. Additionally, verification based on complete chloroplast genomes was performed and new chloroplast markers were developed and evaluated. The combination of these methods resulted in the successful identification of all cultivated Bupleurum individuals. Three chloroplast regions are recommended as additional barcodes for the genus: ycf4_cemA, psaJ_rpl33, and ndhE_ndhG. This is a reliable and promising strategy that can be applied to the authentication of natural products and the identification of other medicinal plant species with similar taxonomic problems.

Goji (fruits of Lycium spp.) is commonly consumed as food and medicine. The increasing market demand for goji has led to its wide cultivation and broad breeding, which might cause loss of genetic diversity. This study aims to uncover the genetic diversity of the cultivated and wild Lycium. The chloroplast genome (CPG) of 34 accessions of Chinese food-medicinal Lycium spp., including the popular cultivars and their wild relatives, was re-sequenced and assembled, based on which the genetic diversity was evaluated. Sequence structural comparison shows that CPG is comparatively conserved within species. Phylogenetic analysis indicates that CPG is sufficient for the discrimination of Lycium species; combined with nuclear ribosomal internal transcribed spacer (Nr ITS) sequences, materials with mixed genetic backgrounds can be identified. Nucleotide diversity analysis reveals that the modern cultivars are probably with a common maternal parent, while the wild accessions are with higher level of genetic diversity. For the first time this study reveals the intraspecies genetic diversity of Lycium spp. using CPG, highlighting the urgent conservation demand of wild genetic resources of Lycium. Our study also demonstrates that CPG provides crucial evidence for identification of Lycium species with mixed genetic backgrounds and highlights the importance of the wild relatives in genetic diversity conservation. This CPG-based technology will contribute to the sustainable development of medicinal plants broadly.

Perilla frutescens (L.) Britton is a highly valuable medicinal plant known for a wide variety of chemotypes based on the components of its essential oils. The appropriate and secure medicinal use of P. frutescens has been severely limited due to the lack of chemotype classification standards and pharmacological studies. In this paper, a classification standard for essential oils in P. frutescens was proposed. The pharmacological activities of different chemotypes or their main components were summarized. Genetic and molecular studies related to the formation of different chemotypes were also reviewed. The molecular mechanism underlying the formation of P. frutescens chemotypes is also discussed to pave the way for the innovation of P. frutescens germplasms for medicinal use.

As a Chinese national key protected medicinal fern naturally occurring in forest understories, Cibotium barometz faces severe threats of wild population degradation, while standardized large-scale artificial breeding technology for conservation purposes remains immature. To establish an efficient spore-based conservation propagation system for this endangered forest fern, this study quantified the independent and interactive effects of spore storage temperature, storage duration and sowing density on spore germination, gametophyte growth and sporophyte seedling establishment. Spores were preserved under four gradient temperature treatments with sequential sampling at multiple storage durations, followed by sowing trials with a series of density gradients; germination rate, seedling establishment rate and gametophyte–sporophyte conversion rate were dynamically recorded and statistically analyzed. The results demonstrated that appropriately extended storage significantly shortened the germination phase and simultaneously elevated both spore germination and sporophyte seedling formation rates. Among all temperature treatments, storage at −4 °C achieved the maximum germination and seedling establishment capacity, whereas ultra-low-temperature cryopreservation at −196 °C greatly promoted gametophyte–sporophyte conversion rate. The optimal sowing density balancing growth space and survival rate was determined to be 30 spores per cm². The complete dynamic developmental traits covering the full spore propagation life cycle of C. barometz were systematically summarized in this work. Our findings supply reliable technical parameters to standardize spore breeding protocols, and offer critical support for ex situ conservation, wild forest population restoration and sustainable resource utilization of C. barometz.

Accurate identification of Fritillaria taxa is essential for quality control of Fritillariae Cirrhosae Bulbus. This study evaluated morphological analysis, chloroplast DNA barcoding, and steroidal alkaloid profiling across 40 individuals from four taxa (F. unibracteata, F. unibracteata var. wabuensis, F. cirrhosa, and F. taipaiensis) from Sichuan province, China. Among five chloroplast markers, ycf1 and rps4 were the most polymorphic, with nucleotide diversity of 0.065 to 0.116 and 0.035 to 0.120, respectively, and generally distinguished the four taxa into well-supported clades, though two F. unibracteata nested within the F. cirrhosa group. Morphologically, F. unibracteata var. wabuensis produced the largest bulbs, with a mean fresh weight of 26.99 g, approximately 24-fold greater than that of F. unibracteata at 1.12 g, whereas F. unibracteata and F. cirrhosa overlapped substantially in all measured traits. PCA revealed that biomass traits were slightly more discriminative, while central bud height contributed the least. Among the samples analyzed, alkaloid profiles provided the most distinct diagnostic signals: F. unibracteata var. wabuensis accumulated the highest sipeimine at 298.40 μg/g and sipeimine-3-β-D-glucoside at 156.30 μg/g; F. taipaiensis was richest in peimine and peiminine; F. unibracteata uniquely contained sipeimine-3-β-D-glucoside without detectable sipeimine; and F. cirrhosa showed the lowest levels across all five compounds. The integration of chloroplast markers with alkaloid profiling improved species discrimination for the samples examined. Chloroplast markers for initial screening followed by the alkaloid profiling of the ambiguous sample might offer a practical framework for the authentication and quality control of Fritillariae Cirrhosae Bulbus, but this strategy requires validation across broader geographic scales and with larger sample sizes.