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Medicinal plants have a variety of values and are an important source of new drugs and their lead compounds. They have played an important role in the treatment of cancer, AIDS, COVID-19 and other major and unconquered diseases. However, there are problems such as uneven quality and adulteration. Therefore, it is of great significance to find comprehensive, efficient and modern technology for its identification and evaluation to ensure quality and efficacy. In this study, deep learning, which is superior to conventional identification techniques, was extended to the identification of the part and region of the medicinal plant Paris polyphylla var. yunnanensis from the perspective of spectroscopy. Two pattern recognition models, partial least squares discriminant analysis (PLS-DA) and support vector machine (SVM), were established, and the overall discrimination performance of the three types of models was compared. In addition, we also compared the effects of different sample sizes on the discriminant performance of the models for the first time to explore whether the three models had sample size dependence. The results showed that the deep learning model had absolute superiority in the identification of medicinal plant. It was almost unaffected by factors such as data type and sample size. The overall identification ability was significantly better than the PLS-DA and SVM models. This study verified the superiority of the deep learning from examples, and provided a practical reference for related research on other medicinal plants.

Jasmonic acids, their effects on plant resistance and metabolites, have been extensively researched. However, methyl dihydrojasmonate (MDJ), a cost-effective and safe substitute for jasmonic acids, lacks sufficient research. This study presents a comprehensive analysis the impact of MDJ on Paris polyphylla var. yunnanensis. The present study aimed to enhance the accumulation of steroidal saponins (polyphyllin I, II, III, D, V, VI, VII, H) in Paris polyphylla var . yunnanensis which is a medicinal plant that contains medicinal steroidal saponins. To achieve this, different concentrations of MDJ were applied to Paris polyphylla var . yunnanensis . The study examined the impact of MDJ on physiology and polyphyllins. The optimal treatment concentration was identified, and the effects of MDJ on physiological metabolism and the expression pattern of related genes were explored. The results indicated that MDJ treatment significantly increased the levels of photosynthetic pigments, antioxidant enzymes, osmoregulators peaked at M2. Moreover, MDJ enhanced the accumulation of polyphyllins in both the leaves and rhizomes peaked at M2, particularly the sum of four polyphyllins (polyphyllin I, II, VI, VII) stated in the Chinese Pharmacopoeia, which increased by 45.65% following treatment. Based on transcriptome analysis, we revealed the polyphyllins-related genes by WGCNA. Furthermore, the majority of genes related to steroidal saponins were up-regulated following MDJ treatment. In the future, we recommend treating Paris polyphylla var . yunnanensis with suitable MDJ concentrations in production to efficiently enhance the yield of steroidal saponins.

Paris yunnanensis , also named as Rhizoma Paridis in the Chinese Pharmacopeia, is a perennial Chinese medicinal herb commonly grown in Southwest China. However, several viruses have been found infecting this plant in recent years. Using high-throughput sequencing (HTS) and Sanger sequencing, this study obtained the complete genome sequences of three capillovirus isolates and one potyvirus isolate. Genomic and phylogenetic analyses revealed that these three capillovirus isolates are the same virus as the newly reported capillovirus, Paris polyphylla chlorotic mottle virus. The newly found potyvirus isolate shares 52.4–68.9% nucleotide sequence identity with other known potyviruses and thus, is grouped into the bean common mosaic virus subgroup. Based on the nucleotide sequence identity, we consider this virus a novel potyvirus species and propose ‘Paris potyvirus 5’ (ParPV-5) as its common name, and ‘ Potyvirus shilinense ’ as its species name. To characterize their biological features, two infectious clones, representing the two viruses, have been constructed through homologous recombination or yeast homologous recombination, and inoculated to several species plants, respectively. The results showed both of the viruses can infect P. yunnanensis and Nicotiana benthamiana . In addition, Paris polyphylla chlorotic mottle virus (PpCMV) can infect N. tabacum var. Xanthi nc, Cucurbita moschata , and Capsicum annuum, and ParPV-5 can infect Cucumis sativus L. and Bidens pilosa L . However, except mild leaf deformation exhibited on the PpCMV-inoculated C. moschata plants, no obvious symptom were observed in these plants including P. yunnanensis . A total of 179 field P. yunnanensis leaf samples from four counties in 2020–2021, and all 640 P. yunnanensis plants from a whole study plot of Lijiang in 2024, were tested using RT-PCR and specific primers, the results showed that PpCMV is a potential preponderant species in some regions, and ParPV-5 has the possible transmission from the original site to other regions.

Background Polyphyllins are significant medicinal compounds found in Paris species, with different polyphyllins fulfilling distinct medicinal roles. Although some genes involved in polyphyllin synthesis have been identified, further exploration of the genes in the polyphyllin synthesis pathway is necessary due to the extensive genome of Paris species. The content and composition of polyphyllins vary among different Paris species, and the variations in specific polyphyllin levels across these plants make them promising candidates for identifying metabolites and genes associated with the biosynthesis of specific polyphyllins. Results In this study, we investigate the global metabolic and transcriptomic profiles of three types of Paris polyphylla var. yunnanensis (Franch.) Hand.-Mazz, one Paris fargesii Franch, and one Paris forrestii (Takht.) H. Li. The rhizome of P. polyphylla is rich in polyphyllin I and II, while P. forrestii is abundant in polyphyllin III, and P. fargesii contains high levels of polyphyllin VI, VII and H. The three Paris species exhibit distinct metabolomic and transcriptomic profiles. Through an integrated analysis of metabolic and transcriptomic data, along with a phylogenetic analysis of genes related to polyphyllin synthesis in Pari s, we annotated a total of six 2,3-oxidosqualene cyclases (OSCs), 120 cytochrome P450s (CYPs), and 138 UDP glycosyltransferases (UGTs). Phylogenetic tree analysis of the obtained data assisted in refining the candidate gene pool for OSC , CYP , and UGT . Subsequently, we identified 6, 12, and 26 candidate genes for OSC , CYP , and UGT , respectively. Finally, by combining the analyses of metabolic and genetic differences, we identified a total of 17 candidate genes, including 2 CAS , 4 CYP , and 11 UGT . Conclusions P. fargesii and P. forrestii are candidate medicinal plants for the development and application of specific polyphyllins. Transcripts from the UGT91 subfamily in Paris may play dual roles, contributing to both the synthesis of polyphyllin II and the catabolism of polyphyllin V and VI. The homologous genes of PpUGT73CE1 may regulate the synthesis of polyphyllin VI in P. fargesii . This study provides new insights into the investigation of biosynthetic pathways in medicinal plants that lack gene clusters.

The [ Paris polyphylla Smith var. Yunnanensis (Franch.) Hand. - Mazz] (The following is denoted by P. polyphylla ) contains various chemical components such as steroidal saponins, flavonoids, polysaccharides, amino acids, etc. It is a traditional Chinese medicinal herb with important medicinal value. So far, the pharmacological research on P. polyphylla at home and abroad mainly focuses on compounds such as saponins and flavonoids, and there are relatively few reports on the study of P. polyphylla polysaccharides. In recent years, with the continuous deepening of research on the P. polyphylla , scientific and technological workers have gradually realized the application value of the polysaccharides from P. polyphylla , and the research of it has been increasing year by year. This article provides a review of the preparation, structural characterization, bioactivities, and potential clinical applications of the polysaccharides from P. polyphylla , aiming to provide a reference for the development, application, and further research.

Background  Paris polyphylla Sm. is a precious medicinal plant rich in various active ingredients. In addition to the well-known saponins, the flavonoids it contains have unique pharmacological potential in antioxidant, neuroprotective, and metabolic regulation. However, the flavonoids in Paris polyphylla Sm. have not been fully researched and developed yet. In this work, we conducted a comprehensive metabolomics and transcriptomics analysis to reveal the metabolic differences and biosynthetic mechanisms of flavonoids in the leaves, stems, and roots of Paris polyphylla Sm. Results Non-targeted metabolomics analysis detected a total of 332 metabolites in Paris polyphylla Sm., among which flavonoids accounted for 19.49%. The diversity and abundance of flavonoids in leaves are the highest, followed by stems and roots. By comparing the metabolites of the roots, stems, and leaves in Paris polyphylla Sm., it was found that there were 45 differential metabolites (DMs) between the leaves and roots, of which flavonoids accounted for 35%. There are 38 DMs between leaves and stems, of which flavonoids account for 45.45%. And there are 52 DMs in stems and roots, among which flavonoids account for 25.53%. A total of 62,766 genes were detected by transcriptomics, and pairwise comparison showed that there were tens of thousands of differentially expressed genes (DEGs) between each group. Afterwards, we selected 39 flavonoids and related metabolites (e.g., kaempferol-3-O-glucoside, quercetin 3-β-D-glucoside, rutin) for targeted metabolomics validation and performed RT-qPCR validation on 29 key flavonoid synthesis genes (e.g., C4H, CHS, FLS, F3’H) to verify the reliability of non-targeted metabolomics and transcriptomics. Conclusions This work indicated that leaves are the main site for the biosynthesis of flavonoids in Paris polyphylla Sm. Among them, kaempferol-3-O-glucoside, quercetin 3-β-D-glucoside, rutin, and other flavonoids are present in higher contents in leaves ( P  < 0.05). Further research on its biosynthetic mechanism indicates that naringenin chalcone is converted to naringenin by chalcone isomerase (CHI). Among them, CHI may be the rate-limiting enzyme in the biosynthesis of flavonoids in Paris polyphylla Sm. The expression of FLS is higher in leaves ( P  < 0.05) and tends to promote the synthesis of flavonols. This work promotes the utilization of non-medicinal parts of Paris polyphylla Sm. and enhances the sustainable development of this precious traditional Chinese medicine resource.

The complete genome sequence of a new potyvirus from Paris polyphylla var. yunnanensis was determined. Its genomic RNA consists of 9571 nucleotides (nt), excluding the 3’-terminal poly(A) tail, containing the typical open reading frame (ORF) of potyviruses and encoding a putative large polyprotein of 3061 amino acids. The virus shares 54.20%-59.60% nt sequence identity and 51.80%-57.90% amino acid sequence identity with other potyviruses. Proteolytic cleavage sites and conserved motifs of potyviruses were identified in the polyprotein and within individual proteins. Phylogenetic analysis indicated that the virus was most closely related to lily yellow mosaic virus. The results suggest that the virus should be classified as a member of a novel species within the genus Potyvirus, and we have tentatively named this virus “Paris yunnanensis mosaic chlorotic virus” (PyMCV).

Paris polyphylla saponin II (PPII) has good biological activity in inhibiting tumor angiogenesis. However, the mechanism of its action is still unclear. This study first observed the inhibitory effect of PPII on cervical cancer cells (Hela) through the establishment of MTT and nude mouse subcutaneous transplantation tumor models. Afterwards, then, we collected Hela cell supernatant for culturing HUVEC cells and treated it with PPII. Observe the invasion, migration, and lumen formation ability of drugs through Transwell, cell scratch test, and angiogenesis experiment. MDC staining was used to observe positive staining in the perinuclear area, AO staining was used to observe acidic areas, and transmission electron microscopy staining was used to observe ultrastructure and autophagy. In addition, the effects of PPII on autophagy- and angiogenesis-related protein expression were detected by Western blotting and quantitative reverse transcriptase polymerase chain reaction. Finally, HUVECs were treated with autophagy inhibitors 3-MA, CQ, and PI3K inhibitor LY294002, respectively. The results showed that the autophagy level of cells treated with PPII was significantly increased. In addition, adding autophagy inhibitors can effectively inhibit angiogenesis in cervical cancer. Further research suggests that PPII induces autophagy in HUVEC cells by regulating the PI3K/AKT/mTOR signaling pathway, thereby affecting angiogenesis and inhibiting Hela cell proliferation, lumen formation, invasion, and migration.

Background Paris polyphylla var. yunnanensis is an important medicinal plant, and the main active ingredient of the plant is polyphyllin, which is a steroid saponin with pharmacological activities. The central enzyme genes participating in the biosynthesis of polyphyllin are increasingly being uncovered; however, UGTs are rarely illustrated. Methods and results In this study, we cloned a new sterol glycosyltransferase from Paris polyphylla var. yunnanensis and identified its catalytic function in vitro. Pp UGT6 showed the ability to catalyse the C-3 glycosylation of pennogenin sapogenin of polyphyllin, and Pp UGT6 showed catalytic promiscuity towards steroids at the C-17 position of testosterone and methyltestosterone and the triterpene at the C-3 position of glycyrrhetinic acid. Homology modelling of the Pp UGT6 protein and virtual molecular docking of Pp UGT6 with sugar acceptors and donors were performed, and we predicted the key residues interacting with ligands. Conclusions Here, Pp UGT6, a novel sterol glycosyltransferase related to the biosynthesis of polyphyllin from P. polyphylla, was characterized . Pp UGT6 catalysed C-3 glycosylation to pennogenin sapogenin of polyphyllin, which is the first glycosylation step of the biosynthetic pathway of polyphyllins. Interestingly, Pp UGT6 demonstrated glycodiversification to testosterone and methyltestosterone at C-17 and triterpene of glycyrrhetinic acid at the C-3 position. The virtual molecular docking of Pp UGT6 protein with ligands predicted the key residues interacting with them. This work characterized a novel SGT glycosylating pennogenin sapogenin at C-3 of polyphyllin from P. polyphylla and provided a reference for further elucidation of the phytosterol glycosyltransferases in catalytic promiscuity and key residues interacting with substrates.

Background: Cancer is one of the top two leading causes of death worldwide. Ethnobotanical research, it is one of methods, which is able to discover effective anticancer drugs based on “prototype” of indigenous people’s historical experiences and practices. The rhizomes of Paris polyphylla var. yunnanensis (Franch.) Hand.-Mazz. have been used as botanical drugs to treat cancer by Yi, Bai, Dai, and Naxi ethnic groups in Yunnan, China, where this species is widely cultivated in a large scale in Yunnan. Materials and methods: To identify the substances of anticancer activities based on indigenous medicine knowledge, chromatography was performed to separate saponins from the rhizomes of P. polyphylla var. yunnanensis , followed by spectroscopy to determine the structure of six isolated saponins. The cytotoxicity of five extracts and six pure compounds were evaluated by MTS method. Quantitative determination of total saponins of P. polyphylla var. yunnanensis was analyzed by HPLC. Cell cycle assay, apoptosis assay, and mitochondrial membrane potential were used to evaluate the pro-apoptotic activity in vitro . Results: Five extracts and six pure saponins showed significant inhibitory cytotoxic activities of three human liver cancer cell lines (SMMC-7721, HepG2, and SK-HEP-1) and one non-small-cell lung cancer cell line (A549). The contents of Paris saponins I, II, and VII were 6.96% in the rhizomes of P. polyphylla var. yunnanensis , much higher than Chinese Pharmacopoeia standards (0.6%). Six saponins induced significant apoptosis and cell cycle arrest in three human cancer cell lines (A549, SMMC-7721, and HepG2), which was associated with the loss of mitochondrial membrane potential. Conclusion: The result of this study support that cultivated P. polyphylla var. yunnanensis could be a substitute for wild resource as an anticancer medicine based on indigenous medicine knowledge.