Cepharanthine analogs mining and genomes of Stephania accelerate anti-coronavirus drug discovery.

Cepharanthine is a secondary metabolite isolated from Stephania. It has been reported that it has anti-conronaviruses activities including severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Here, we assemble three Stephania genomes (S. japonica, S. yunnanensis, and S. cepharantha), propose the cepharanthine biosynthetic pathway, and assess the antiviral potential of compounds involved in the pathway. Among the three genomes, S. japonica has a near telomere-to-telomere assembly with one remaining gap, and S. cepharantha and S. yunnanensis have chromosome-level assemblies. Following by biosynthetic gene mining and metabolomics analysis, we identify seven cepharanthine analogs that have broad-spectrum anti-coronavirus activities, including SARS-CoV-2, Guangxi pangolin-CoV (GX_P2V), swine acute diarrhoea syndrome coronavirus (SADS-CoV), and porcine epidemic diarrhea virus (PEDV). We also show that two other genera, Nelumbo and Thalictrum, can produce cepharanthine analogs, and thus have the potential for antiviral compound discovery. Results generated from this study could accelerate broad-spectrum anti-coronavirus drug discovery.

Comparative analysis of medicinal plants Scutellaria baicalensis and common adulterants based on chloroplast genome sequencing.

BACKGROUND: Scutellaria baicalensis Georgi has been extensively used as a medicinal herb in China for over 2000 years. They may be intentionally or inadvertently substituted or blended with comparable species in the local market, threatening clinical medication safety. Molecular markers are effective tools to prevent misidentification and eliminate doping and falsification among Scutellaria plants. This study screened four highly variable regions to identify Scutellaria and its adulterants. In addition, a phylogenetic analysis was performed using the complete cp genome combined with published Scutellaria species samples. Moreover, a comparative analysis of the cp genomes was conducted to investigate the cp genome evolution of S. baicalensis. RESULTS: The complete cp genome of five species of Scutellaria was sequenced for the first time, and four previously published Scutellaria species were re-sequenced. They all exhibited a conserved quadripartite structure in their cp genomes, including two distinct regions, namely a small and large single copy region, respectively, and two inverted repeats encompassing the majority of ribosomal RNA genes. Furthermore, the nine species exhibited high conservation from aspects of the genome structure, codon usage, repeat sequences, and gene content. Four highly variable regions (matK-rps16, ndhC-trnV-UAC, psbE-petL, and rps16-trnQ-UUG) may function as potential molecular markers for differentiating S. baicalensis from its adulterants. Additionally, the monophyly of Scutellaria was ascertained and could be reclassified into two subgenera, subgenus Anaspis and subgenus Scutellaria, as evidenced by the phylogenetic analyses on sequences of cp genome and shared protein-coding sequences. According to the molecular clock analysis, it has been inferred that the divergence of Scutellaria occurred at approximately 4.0 Mya during the Pliocene Epoch. CONCLUSION: Our study provides an invaluable theoretical basis for further Scutellaria species identification, phylogenetics, and evolution analysis.

Extraction, purification, characteristics, bioactivities, prospects, and toxicity of Lilium spp. polysaccharides.

The genus Lilium has been widely used worldwide as a food and medicinal ingredient in East Asia for over 2000 years due to its higher nutritional and medicinal value. Polysaccharide is the most important bioactive ingredient in Lilium spp. and has various health benefits. Recently, Lilium spp. polysaccharides (LSPs) have attracted significant attention from industries and researchers due to their various biological properties, such as antioxidant, immunomodulatory, antitumor, antibacterial, hypoglycaemic, and anti-radiation. However, the development and utilization of LSP-based functional biomaterials and medicines are limited by a lack of comprehensive understanding regarding the structure-activity relationships (SARs), industrial applications, and safety of LSPs. This review provides an inclusive overview of the extraction, purification, structural features, bioactivities, and mechanisms of LSPs. SARs, applications, toxicities, and influences of structural modifications on bioactivities are also highlighted, and the potential development and future study direction are scrutinized. This article aims to offer a complete understanding of LSPs and provide a foundation for further research and application of LSPs as therapeutic agents and multifunctional biomaterials.

Comparatively analyzing of chloroplast genome and new insights into phylogenetic relationships regarding the genus Stephania.

The medicinal plant of the genus Stephania holds significant economic importance in the pharmaceutical industry. However, accurately classifying and subdividing this genus remains a challenge. Herein, the chloroplast (cp) genomes of Stephania and Cyclea were sequenced, and the primary characteristics, repeat sequences, inverted repeats regions, simple sequence repeats, and codon usage bias of 17 species were comparatively analyzed. Twelve markers were identified through genome alignment and sliding window analysis. Moreover, a molecular clock analysis revealed the divergence between subgenus (subg.) Botryodiscia and the combined Cyclea, subg. Stephania and Tuberiphania during the early Oligocene epoch. Notably, the raceme-type inflorescence represents the ancestral state of the Stephania and Cyclea. The genetic relationships inferred from the cp genome and protein-coding genes exhibited similar topologies. Additionally, the paraphyletic relationship between the genera Cyclea and Stephania was confirmed. Bayesian inference, maximum likelihood, and neighbor-joining trees consistently showed that section Tuberiphania and Transcostula were non-monophyletic. In conclusion, this research provides valuable insights for further investigations into species identification, evolution, and phylogenetics within the Stephania genus.

Comparative analysis of chloroplast genome and new insights into phylogenetic relationships of Ajuga and common adulterants.

Introduction: The potential contamination of herbal medicinal products poses a significant concern for consumer health. Given the limited availability of genetic information concerning Ajuga species, it becomes imperative to incorporate supplementary molecular markers to enhance and ensure accurate species identification. Methods: In this study, the chloroplast (cp) genomes of seven species of the genus Ajuag were sequenced, de novo assembled and characterized. Results: exhibiting lengths ranging from 150,342 bp to 150,472 bp, encompassing 86 - 88 protein-coding genes (PCGs), 35 - 37 transfer RNA, and eight ribosomal RNA. The repetitive sequences, codon uses, and cp genomes of seven species were highly conserved, and PCGs were the reliable molecular markers for investigating the phylogenetic relationship within the Ajuga genus. Moreover, four mutation hotspot regions (accD-psaI, atpH-atpI, ndhC-trnV(UAC), and ndhF-rpl23) were identified within cp genomes of Ajuga, which could help distinguish A. bracteosa and its contaminants. Based on cp genomes and PCGs, the phylogenetic tree preliminary confirmed the position of Ajuga within the Lamiaceae family. It strongly supported a sister relationship between Subsect. Genevense and Subsect. Biflorae, suggesting the merger of Subsect. Biflorae and Subsect. Genevenses into one group rather than maintaining separate categorizations. Additionally, molecular clock analysis estimated the divergence time of Ajuga to be around 7.78 million years ago. Discussion: The species authentication, phylogeny, and evolution analyses of the Ajuga species may benefit from the above findings.

Analysis of codon usage patterns in 48 Aconitum species.

BACKGROUND: The Aconitum genus is a crucial member of the Ranunculaceae family. There are 350 Aconitum species worldwide, with about 170 species found in China. These species are known for their various pharmacological effects and are commonly used to treat joint pain, cold abdominal pain, and other ailments. Codon usage bias (CUB) analysis contributes to evolutionary relationships and phylogeny. Based on protein-coding sequences (PCGs), we selected 48 species of Aconitum for CUB analysis. RESULTS: The results revealed that Aconitum species had less than 50% GC content. Furthermore, the distribution of GC content was irregular and followed a trend of GC1 > GC2 > GC3, indicating a bias towards A/T bases. The relative synonymous codon usage (RSCU) heat map revealed the presence of conservative codons with slight variations within the genus. The effective number of codons (ENC)-Plot and the parity rule 2 (PR2)-bias plot analysis indicate that natural selection is the primary factor influencing the variation in codon usage. As a result, we screened various optimal codons and found that A/T bases were preferred as the last codon. Furthermore, our Maximum Likelihood (ML) analysis based on PCGs among 48 Aconitum species yielded results consistent with those obtained from complete chloroplast (cp.) genome data. This suggests that analyzing mutation in PCGs is an efficient method for demonstrating the phylogeny of species at the genus level. CONCLUSIONS: The CUB analysis of 48 species of Aconitum was mainly influenced by natural selection. This study reveals the CUB pattern of Aconitum and lays the foundation for future genetic modification and phylogenetic analyses.

The potential, challenges, and prospects of the genus Spirulina polysaccharides as future multipurpose biomacromolecules.

Spirulina has been widely used worldwide as a food and medicinal ingredient for centuries. Polysaccharides are major bioactive constituents of Spirulina and are of interest because of their functional properties and unlimited application potential. However, the clinical translation and market industrialization of the polysaccharides from genus Spirulina (PGS) are retarded due to the lack of a further understanding of their isolation, bioactivities, structure-activity relationships (SARs), toxicity, and, most importantly, versatile applications. Herein, we provide an overview of the extraction, purification, and structural features of PGS; meanwhile, the advances in bioactivities, SARs, mechanisms of effects, and toxicity are discussed and summarized. Furthermore, the applications, potential developments, and future research directions are scrutinized and highlighted. This review may help fill the knowledge gap between theoretical insights and practical applications and guide future research and industrial application of PGS.

Preparation, structural characterization, bioactivities, and applications of Crataegus spp. polysaccharides: A review.

Crataegus, is a genus within the Rosaceae family. It is recognized as a valuable plant with both medicinal and edible qualities, earning it the epithet of the "nutritious fruit" owing to its abundant bioactive compounds. Polysaccharides are carbohydrate polymers linked by glycosidic bonds, one of the crucial bioactive ingredients of Crataegus spp. Recently, Crataegus spp. polysaccharides (CPs) have garnered considerable attention due to their diverse range of bioactivities, including prebiotic, hypolipidemic, anticancer, antibacterial, antioxidant, and immunobiological properties. Herein, we provide a comprehensive overview of recent research on CPs. The analysis revealed that CPs exhibited a broad molecular weight distribution, ranging from 5.70 Da to 4.76 × 108 Da, and are composed of various monosaccharide constituents such as mannose, rhamnose, and arabinose. Structure-activity relationships demonstrated that the biological function of CPs is closely associated with their molecular weight, galacturonic acid content, and chemical modifications. Additionally, CPs have excellent bioavailability, biocompatibility, and biodegradability, which make them promising candidates for applications in the food, medicine, and cosmetic industries. The article also scrutinized the potential development and future research directions of CPs. Overall, this article provides comprehensive knowledge and underpinnings of CPs for future research and development as therapeutic agents and multifunctional food additives.

Bioactive polysaccharides from Momordica charantia as functional ingredients: a review of their extraction, bioactivities, structural-activity relationships, and application prospects.

Momordica charantia L. is a well-known medicine and food homology plant with high pharmaceutical and nutritional values. Polysaccharides are carbohydrate polymers connected by glycosidic bonds, one of the key functional ingredients of M. charantia. Recently, M. charantia polysaccharides (MCPs) have attracted much attention from industries and researchers due to their anti-oxidant, anti-tumor, anti-diabetes, anti-bacteria, immunomodulatory, neuroprotection, and organ protection activities. However, the development and utilization of MCPs-based functional foods and medicines were hindered by the lack of a deeper understanding of the structure-activity relationship (SAR), structural modification, applications, and safety of MCPs. Herein, we provide an overview of the extraction, purification, structural characterization, bioactivities, and mechanisms of MCPs. Besides, SAR, toxicities, application, and influences of the modification associated with bioactivities are spotlighted, and the potential development and future study direction are scrutinized. This review provides knowledge and research underpinnings for the further research and application of MCPs as therapeutic agents and functional food additives.

Bioactive polysaccharides from lotus as potent food supplements: a review of their preparation, structures, biological features and application prospects.

Lotus is a famous plant of the food and medicine continuum for millennia, which possesses unique nutritional and medicinal values. Polysaccharides are the main bioactive component of lotus and have been widely used as health nutritional supplements and therapeutic agents. However, the industrial production and application of lotus polysaccharides (LPs) are hindered by the lack of a deeper understanding of the structure-activity relationship (SAR), structural modification, applications, and safety of LPs. This review comprehensively comments on the extraction and purification methods and structural characteristics of LPs. The SARs, bioactivities, and mechanisms involved are further evaluated. The potential application and safety issues of LPs are discussed. This review provides valuable updated information and inspires deeper insights for the large scale development and application of LPs.

Extraction, structural-activity relationships, bioactivities, and application prospects of Bletilla striata polysaccharides as ingredients for functional products: A review.

Bletilla striata is a well-known medicinal plant with high pharmaceutical and ornamental values. Polysaccharide is the most important bioactive ingredient in B. striata and has various health benefits. Recently, B. striata polysaccharides (BSPs) have attracted much attention from industries and researchers due to their remarkable immunomodulatory, antioxidant, anti-cancer, hemostatic, anti-inflammatory, anti-microbial, gastroprotective, and liver protective effects. Despite the successful isolation and characterization of BSPs, there is still limited knowledge regarding their structure-activity relationships (SARs), safety concerns, and applications, which hinders their full utilization and development. Herein, we provided an overview of the extraction, purification, and structural features, as well as the effects of different influencing factors on the components and structures of BSPs. We also highlighted and summarized the diversity of chemistry and structure, specificity of biological activity, and SARs of BSP. The challenges and opportunities of BSPs in the food, pharmaceutical, and cosmeceutical fields are discussed, and the potential development and future study direction are scrutinized. This article provides comprehensive knowledge and underpinnings for further research and application of BSPs as therapeutic agents and multifunctional biomaterials.

Extraction, structural-activity relationships, bioactivities, and application prospects of Pueraria lobata polysaccharides as ingredients for functional products: A review.

Pueraria lobata (Willd.) Ohwi is an important resource with dual functions in medicine and food since ancient times. Polysaccharides are the main bioactive component of P. lobata and have various bioactivities, such as antidiabetic, antioxidant, immunological activities, etc. Due to the distinctive bioactivity of P. lobata polysaccharides (PLPs), the research on PLPs is booming. Although a series of PLPs have been isolated and characterized, the chemical structure and mechanism are unclear and need further study. Here, we reviewed recent progress in isolation, identification, pharmacological properties, and possible therapeutic mechanisms of PLPs to update awareness of these value-added natural polysaccharides. Besides, the structure-activity relationships, application status, and toxic effects of PLPs are highlighted and discussed to afford a deeper understanding of PLPs. This article may provide theoretical insights and technical guidance for developing PLPs as novel functional foods.

Comparative analysis of medicinal plant Isodon rubescens and its common adulterants based on chloroplast genome sequencing.

Isodon rubescens (Hemsley) H. Hara is the source of Donglingcao under the monograph Rabdosiae Rubescentis Herba in Chinese Pharmacopoeia. In the local marketplace, this medicine can be accidentally contaminated, deliberately substituted, or mixed with other related species. The contaminants of herbal products are a threat to consumer safety. Due to the scarcity of genetic information on Isodon plants, more molecular markers are needed to avoid misidentification. In the present study, the complete chloroplast (cp) genome of seven species of Isodon was sequenced, de novo assembled and characterized. The cp genomes of these species universally exhibited a conserved quadripartite structure, i.e., two inverted repeats (IRs) containing most of the ribosomal RNA genes and two unique regions (large single copy and small single copy). Moreover, the genome structure, codon usage, and repeat sequences were highly conserved and showed similarities among the seven species. Five highly variable regions (trnS-GCU-trnT-CGU, atpH-atpI, trnE-UUC-trnT-GGU, ndhC-trnM-CAU, and rps15-ycf1) might be potential molecular markers for identifying I. rubescens and its contaminants. These findings provide valuable information for further species identification, evolution, and phylogenetic research of Isodon.

Phylogenetic relationship and characterization of the complete chloroplast genome of Laggera crispata, a folk herbal medicine plant in China.

Laggera crispata, an herbaceous plant, has been used in Chinese medicines as anti-inflammatory, analgesic, and anti-viral. In this study, the complete chloroplast (cp) genome sequence of L. crispata was first reported. The cp genome of L. crispata is 155522 bp in length, with two inverted repeats (IR) regions of 25042 bp, the large single copy (LSC) region of 84198 bp and the small single copy (SSC) region of 21240 bp. 128 genes were predicted, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic analysis suggested that L. crispata is more closely related to Pluchea pteropoda and P. indica with solid bootstrap values belonging to the subfamily Inuleae of Asteraceae.

Comparative Analysis of Chloroplast Genome and New Insights Into Phylogenetic Relationships of Polygonatum and Tribe Polygonateae.

Members of Polygonatum are perennial herbs that have been widely used in traditional Chinese medicine to invigorate Qi, moisten the lung, and benefit the kidney and spleen among patients. However, the phylogenetic relationships and intrageneric taxonomy within Polygonatum have long been controversial because of the complexity of their morphological variations and lack of high-resolution molecular markers. The chloroplast (cp) genome is an optimal model for deciphering phylogenetic relationships in related families. In the present study, the complete cp genome of 26 species of Trib. Polygonateae were de novo assembled and characterized; all species exhibited a conserved quadripartite structure, that is, two inverted repeats (IR) containing most of the ribosomal RNA genes, and two unique regions, large single sequence (LSC) and small single sequence (SSC). A total of 8 highly variable regions (rps16-trnQ-UUG, trnS-GCU-trnG-UCC, rpl32-trnL-UAG, matK-rps16, petA-psbJ, trnT-UGU-trnL-UAA, accD-psaI, and trnC-GCA-petN) that might be useful as potential molecular markers for identifying Polygonatum species were identified. The molecular clock analysis results showed that the divergence time of Polygonatum might occur at ∼14.71 Ma, and the verticillate leaf might be the ancestral state of this genus. Moreover, phylogenetic analysis based on 88 cp genomes strongly supported the monophyly of Polygonatum. The phylogenetic analysis also suggested that Heteropolygonatum may be the sister group of the Polygonatum, but the Disporopsis, Maianthemum, and Disporum may have diverged earlier. This study provides valuable information for further species identification, evolution, and phylogenetic research of Polygonatum.

Comparative analysis of complete chloroplast genome sequences of five endangered species and new insights into phylogenetic relationships of Paris.

Paris L. genus has been a precious traditional herb for more than 2000 years in China. However, due to overexploitation and habitat destruction, Paris is threatened by extinction. Similar morphological features cause the classification of Paris species in dispute. The chloroplast (cp) genome approach has been used to investigate the evolution of Paris. However, some studies confirm that the cp genome may result in misleading relationships because of the length variation, gaps/indels deletion, and incorrect models of sequence evolution in concatenated datasets. Therefore, there is a high demand for a reconstructed phylogenetic relationship and developed genetic markers to conserve these species. Recent studies have demonstrated that the protein-coding genes could provide a better phylogenetic relationship in the phylogenetic investigation. In this study, the complete cp genomes of five species were characterized, and the length of five cp genomes ranges from 162,927 bp to 165,267 bp, covering 89 protein-coding genes, 38 tRNA, and eight rRNA. The analysis of the repeat sequences, codon usage, RNA-editing sites, and comparison of cp genomes shared a high degree of conservation. Based on the protein-coding genes, the phylogenetic tree confirmed Paris's position in the order Melanthiaceae, providing maximum support for a sister relationship of the subgenera Paris sensu strict (Paris s.s.) with the Daiswa and Trillium. In addition, the molecular clock showed that subgenus Paris was inferred to have occurred at about 52.81 Mya, whereas subgenus Daiswa has originated at 24.56 Mya, which was consistent with the phylogenetic investigation. This study provided a valuable insight into the evolutionary dynamics of cp genome structure in the family Melanthiaceae, and it also contributes to the bioprospecting and conservation of Paris species.

The first complete chloroplast genome of Vicatia thibetica de Boiss.: genome features, comparative analysis, and phylogenetic relationships.

Vicatia thibetica de Boiss.: a herb in the family Apiaceae, has been used for over a hundred years as an essential medicinal and edible plant in the Bai ethnic group of Dali City. However, due to the lack of study on plastid genomes of V. thibetica, studies of comparison and phylogeny with other related species remain scarce. In the current study, we assembled, annotated, and characterized the entire chloroplast (cp) genome of V. thibetica through high-throughput sequencing for the first time, compared with published whole chloroplast genomes from the same family. A phylogenetic analysis of the chloroplast genome has also been performed. The whole chloroplast genome of V. thibetica was 145,796 in size and consisted of a large single-copy region (LSC; 92,186 bp), a small single-copy region (SSC; 17,452 bp), and a pair of inverted repeat regions (IRs; 18,079 bp) forming a circular quadripartite structure. Annotation resulted in 128 genes, including 84 protein-coding genes (PCGs), 35 transfer RNA genes (tRNAs), eight ribosomal genes (rRNAs), and one pseudogene. Repeat sequence analysis displayed V. thibetica plastid genome contains 75 simple repeats, 37 long repeats, and 29 tandem repeats. Compared with the cp genome of other Apiaceae species, a common feature was that the IR regions of the genome were more conservative compared to the LSC and SSC regions. Highly variable hotspots included rps16, ndhC-trnV-UAC, clpP, ycf1, and ndhB in the genomes, which supply valuable molecular markers for phylogeny, identification, and classification in the Apiaceae family. The results of phylogenetic analysis strongly supported the genus Vicatia as an independent genus in the family Apiaceae, in which the closest affinities to the related species of Angelica, Peucedanum, and Ligusticum were observed. In conclusion, the first chloroplast genome of Vicatia reported in this study may  improve our understanding of phylogenetic relationship of different genera of Apiaceae. In addition, the current data will be valuable as chloroplast genomic resource for species identification and population genetics. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01154-y.

Detection of Gegen Adulteration Using Multiple Fingerprints Coupled With Chemometric Strategy.

BACKGROUND: The root of Pueraria montana var. lobata (gegen) is a well-known traditional Chinese medical herb, which is prone to be accidentally contaminated with Pueraria montana var. thomsonii, Pueraria wallichii, and Pueraria peduncularis due to the morphological character similarity. These adulterations might cause quality confusion and safety issues. OBJECTIVE: In this study, the screening technique to detect adulteration in gegen was developed using multiple fingerprints and chemometrics. METHOD: A range of gegen samples and possible known adulterants including Pueraria montana var. thomsonii, Pueraria wallichii, and Pueraria peduncularis were collected. FT-IR and HPLC coupled with different chemometric techniques, including similarity analysis (SA), hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least-squares discriminant analysis (OPLS-DA), provide the qualitative chemometric models for gegen adulteration detection. RESULTS: FT-IR and HPLC combined with OPLS-DA successfully differentiated authentic gegen from adulterants. Both FT-IR and HPLC units can be used as alternative methods to traditional methods. The HPLC showed better performance in identifying samples than FT-IR. CONCLUSIONS: The use of FT-IR and HPLC coupled with chemometrics could potentially be the proper selection method for the early quality evaluation of gegen. This method can be used to combat fraud in the herbal industry in the future. HIGHLIGHTS: FT-IR and HPLC combined with chemometrics analysis were developed to discriminate between Pueraria lobata (gegen) and adulterants. The multiple fingerprints combined with multivariate methods were successfully applied to the study of the gegen and its adulterants. The chemometrics analysis using SA and OPLS-DA indicate significant differentiation in the chemical composition of these species. This research provides important chemotaxonomic references in species identification.

The complete chloroplast genome of Aconitum piepunense (Ranunculaceae) and its phylogenetic analysis.

Aconitum piepunense belonging to the family Ranunculaceae is an endangered herb species in southwestern China. In this study, the complete chloroplast genome of A. piepunense was sequenced, and the results revealed a typical quadripartite structure with a length of 155,836 bp, including a large single-copy region (LSC, 86,433 bp), a small single-copy region (SSC, 16,945 bp), and a pair of inverted repeat (IR) regions (IRa and IRb, 26,229 bp, respectively). A total of 130 genes were identified in the A. piepunense chloroplast genome, containing 85 protein-coding genes, 37 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. Phylogenetic analysis based on the maximum likelihood method indicated that A. piepunense formed a monophyletic group, which was sister to A. contortum and A. vilmorinianum.