Paris Polyphylla Research Articles

First Report of Colletotrichum spaethianum Causing Anthracnose on Jiaotou (Allium chinense G. Don) in China.

Jiaotou (Allium chinense G. Don) is a culinary and aromatic herb belonging to the family Amaryllidaceae. It is native to China and cultivated for its culinary uses in many other countries, which also possess high medicinal value in traditional Chinese medicine (Rhetso et al. 2020). In March 2023, a disease resembling anthracnose was detected in the fields of Changde city, Hunan Province, China. The symptoms manifested as yellow to brown necrotic spots located at the tip or middle of the blade. The lesions tended to wrinkle, leading to leaf distortion, thinning, and wilting. This disease has a high incidence rate of approximately 50% in an area of nearly 20 hectares. Ten symptomatic plants were collected for pathogen isolation. The leaf tissues were surface sterilized using 70% ethyl alcohol and 2% sodium hypochlorite, rinsed with sterile distilled water, and then transferred to potato dextrose agar (PDA) and incubated at 26°C in the dark. Ten colonies with similar cultural morphology were selected from the 10 affected plant samples. The fungal strains produced densely aerial mycelium on PDA, displaying off-white to gray-brown coloration. Conidia were hyaline, curved or slightly curved, aseptate, with a rounded apex and truncate base, contained oil globules, and measuring 21.0 to 29.0 × 4.1 to 5.3 μm (n = 50. Appressoria were dark brown, irregular in shape, partially lobed, and measured 6.5 to 15.5 µm long × 5.0 to 10.5 µm wide. Setae were straight, dark brown, with two- to three-septate, 78.5 to 155.4 μm long (n = 50). These morphological characteristics were similar to Colletotrichum spaethianum (Damm et al. 2009). The internal transcribed spacer (ITS) region of rDNA, actin (ACT), chitin synthase (CHS-1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes of three representative strains, JT-1, JT-2 and JT-3, were amplified and sequenced as described in Weir et al. (2012). These sequences were 99 to 100% identical to corresponding sequences of C. spaethianum strains (565/569 bp, MN396364.1 for ITS; 565/569 bp, MN433299.1 for ACT; 262/263 bp, MN433298.1 for GAPDH; 272/272 bp, MH020773 for CHS-1, respectively) (Zhong et al. 2020; Liu et al. 2020; Sun et al. 2020). A concatenated phylogenetic tree showed that all three strains were clustered within the C. spaethianum clade. The sequences of isolate JT-1 were deposited in GenBank (accession nos. PP911453, PP911607, PP911605, and PP911606 for ITS, ACT, GAPDH and CHS-1, respectively). For pathogenicity assays, 1-month-old seedings of A. chinense in plots were inoculated by spraying with a conidial suspension (1 × 105 conidia/ml) until runoff. Control plants were sprayed with sterilized water. All the inoculated plants were kept in a greenhouse at 25°C with a 12-h/12-h light/dark cycle. The experiment was conducted twice with three replications each. After 7 days, symptoms similar to those observed in the field developed on leaves of the inoculated plants, whereas the control plants remained asymptomatic C. spaethianum was specifically reisolated and identified from the symptomatic leaves, thus fulfilling Koch's postulates. Colletotrichum spaethianum has been recorded as a pathogen fungus on many medicinal plants, such as Peucedanum praeruptorum (Guo et al. 2013), Anemarrhena asphodeloides (Okorley et al. 2019), and Paris polyphylla (Zhong et al. 2020). To our knowledge, this is the first report of C. spaethianum causing anthracnose on Jiaotou in China. Effective control measures should be implemented to mitigate potential further damage to Jiaotou cultivation.

First Report of Leaf Blight on Paris polyphylla Caused by Ceratobasidium ramicola in China.

Paris polyphylla var. yunnanensisis a perennial herb with significant medicinal properties in anticancer, anti-inflammatory, antibacterial immunomodulatory and antispasmodic activities (Duan et al. 2018). In April 2022, leaf blight disease emerged in Xiangtan City (Hunan), affecting P. polyphylla plantings over an area of 3×104 m2 (27.904°N, 112.918°E). The disease incidence reached an average of 22% of the plants in the field, with infected plants initially displaying water-soaked chlorosis, followed by dry yellow shrinkage that gradually spread from the leaf tips to the entire plant. To identify the causal agent, 20 leaf lesions (4 mm2) collected from 20 plants were surface sterilized with 75% ethanol for 10 s, 5% NaOCl for 30 s, rinsed in sterile distilled water three times, transferred to potato dextrose agar (PDA) plates with lactic acid (0.125%) , and incubated at 28 °C in the dark. Four isolates (PP21 to PP24) with similar morphologies were obtained and purified by the hyphal-tip method. Colonies on PDA initially appeared white with cottony mycelium, later turning light-yellow on the underside. Septate hyphae were branched at right angles with a small constriction at the departure of the branch point and measured 4.16 to 7.93 µm in diameter. Binucleate cells within the septate hyphae were visualized using Giemsa staining (Servicebio, China). For molecular identification, the rDNA internal transcribed spacer (ITS), the second largest subunit of nuclear DNA-directed RNA polymerase II (RPB2), and ATP synthase subunit 6 (ATP6) were amplified from genomic DNA of the isolates extracted by Fungus Genomic DNA Extraction Kit (Bioflux, China) using primers ITS1/ITS4 (White et al. 1990), bRPB2-6F/bRPB2-7.1R (Matheny 2005; Reeb et al. 2004) and ATP61/ATP62 (Kretzer and Bruns 1999), respectively. The ITS, RBP2 and ATP6 of the four isolates were sequenced and deposited in GenBank. BLASTn search of sequenced ITS (PQ187050, PQ187051, PP728052, PQ187052), RBP2 (PQ202833, PQ202834, PP735921, PQ202835) and ATP6 (PQ202837, PQ202838, PP735922, PQ202839) revealed a >99% identity with the type strain of Ceratobasidium ramicola CBS133.82 (NR138368, DQ301708, and DQ301577). For phylogenetic analysis, concatenated sequences of ITS, RBP2, and ATP6 were employed using the maximum-likelihood method in MEGA-X. Based on the morphological and molecular analyses, the isolates were classified into the C. ramicola clade (Bandoni 1979; Samuels et al. 2012; Jeong et al. 2023). To test the pathogenicity of the isolate PP23, mycelial plugs (5 mm in diameter) were placed directly on 15 healthy leaves from 15 three-year-old plants after puncturing with sterile needles. Sterile PDA plugs served as controls. All plants were kept in a greenhouse with conditions of 25°C, 80% relative humidity and a photoperiod of 12 h. After 5 days, all infected plants developed leaf blight symptoms similar to those described above, whereas the control plants remained asymptomatic. The pathogenicity test was conducted three times. C. ramicolawas re-isolated from all symptomatic leaves and confirmed to be identical to the original isolate based on morphology and nucleotide sequences of ITS, RBP2, and ATP6. While C. ramicola is commonly reported to be isolated from diseased cacao, its pathogenicity to cacao remains unknown (Samuels et al. 2012). To our knowledge, this is the first report of C. ramicola causing leaf blight on P. polyphylla, a medicinal herb with significant economic importance in China.

Ethnobotanical and Ethnopharmacological Study of Paris polyphylla var. yunnanensis in Yunnan Province, China

The traditional medicinal knowledge in the northwest of Yunnan Province, China have been poorly studied. Paris polyphylla var. yunnanensis (PPvY) is widely cultivated and used as indigenous traditional Chinese medicine (TCM) to treat cancer in northwest Yunnan. This study aims to reveal the traditional medicinal knowledge of PPvY and folk formulas related to PPvY through literature research and ethnobotanical investigation. Semi-structured interviews were conducted with 14 highly regarded folk doctors in the northwest of Yunnan, China, based on relevant data collected in the initial phase of the research. We identified twenty-three traditional treatments, thirty pairing herbs used with PPvY in therapy, and eight processing methods of PPvY. The results indicated that PPvY and its associated formulas containing PPvY were primarily used for treating cancer and inflammation and for clearing heat and detoxifying. The TCM herbs most frequently used alongside PPvY included Engleromyces sinensis and Glycyrrhiza yunnanensis. The commonly employed processing methods primarily involved using PPvY in both its dry and fresh forms, while special processing methods, such as processing in wine and honey, steaming, and foil-packet boiling, were worth further research. Our results highlight the diversity of medicinal plants and the richness of traditional medical knowledge in northwest Yunnan, China. This study may offer clues for the development and research of indigenous medicinal plants. Additionally, a collective effort is needed to create a plan for the sustainable use of indigenous medicinal plants, enhancing local economic development while safeguarding biodiversity and traditional medicinal knowledge.

Polyphyllin VI Ameliorates Pulmonary Fibrosis by Suppressing the MAPK/ERK and PI3K/AKT Signaling Pathways via Upregulating DUSP6.

Pulmonary fibrosis (PF) is a lethal disease caused by inordinate repair of damaged lungs, for which limited strategies are available. Polyphyllin VI (PPVI), extracted and isolated from Paris polyphylla Smith var. chinensis (Franch.) Hara, has been regarded as an important traditional Chinese herbal medicine for the treatment of respiratory system diseases. This study evaluated effects of PPVI on PF and its underlying mechanism. Experimental procedure For evaluating the anti-PF effect of PPVI, we established an in vivo PF mouse model via intratracheal infusion of bleomycin (BLM) in mice and an in vitro PF model induced by TGF-β1 in NIH/3T3, HPF and A549, respectively. Subsequently, the mechanism of PPVI effects was further explored using RNA sequencing (RNA-Seq). The in vivo and in vitro results demonstrated that PPVI significantly inhibited inflammation, oxidative damage, and epithelial-mesenchymal transition. Furthermore, RNA sequencing indicated that PPVI ameliorated PF by modulating inflammation and oxidative stress responses. Furthermore, dual specificity phosphatase 6 (DUSP6), was the shared and most significant differentially expressed gene associated with inflammation and oxidative stress response after PPVI treatment. Mechanistically, silencing DUSP6 can eliminate the suppressive impact on PPVI for the activation of fibroblast and the phosphorylation of ERK and AKT. Summarily, our findings revealed the potential of PPVI in mitigating PF via upregulating DUSP6 and highlighted the regulatory function of DUSP6 in the pathogenesis of PF.

Understory growth of Paris polyphylla accumulates a reservoir of secondary metabolites of plants.

Paris polyphylla is an important traditional medicinal plant of the Himalayan region. It is extensively used for the production of natural steroidal saponins and flavonoids. Although seed dormancy of wild plants can be broken to be artificially maintained and regenerated through micropropagation in the laboratory, the success of secondary metabolite production in higher quantities and the synthesis of superior plant metabolites have been very limited. In this study, we present differential metabolic profiling of P. polyphylla plants maintained for 8 years in natural and greenhouse conditions. Untargeted profiling of the metabolites through ultra-performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS), followed by statistical analysis, identified secondary metabolites that were enriched in the naturally occurring plant roots compared with the greenhouse plant roots. A multivariate statistical analysis revealed the differential distribution of the compounds between the two groups. Overall, we identified 1,182 secondary metabolites, with 116 metabolites being differentially upregulated and 256 metabolites being downregulated. Moreover, 810 metabolites showed no significant variation under both growing conditions. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the naturally forest-grown P. polyphylla plants were significantly enriched in steroidal saponins, lipids, vitamins, flavonoids, and flavonols. An analysis of the top 10 differentially upregulated secondary metabolites indicated a significantly enriched quantity of spirost-5-en-3,12-diol and kaempferol synthesis pathways, which are known to reduce the effect of free radicals scavengers inside the cell. In addition, veratramine alkaloids were also enriched under natural conditions. Our findings indicated that naturally maintained P. polyphylla plants are suitable for the extraction of medicinally important compounds. Our study established a causal relationship between the metabolic composition of the roots and their natural growth condition. This study highlighted the importance of environmental conditions in the biosynthesis of secondary metabolites of plants.

Identification and pathogenicity analysis of Alternaria alternata, the causal agent of leaf spot disease in Paris polyphylla var. yunnanensis

Identification and pathogenicity analysis of Alternaria alternata, the causal agent of leaf spot disease in Paris polyphylla var. yunnanensis

Investigate growth of Paris polyphylla under synergic effects of CeO2 and SiO2 using as fertilizers

This study successfully synthesized SiO2 and CeO2 nano-materials to fertilize for Paris polyphylla (P. polyphylla). The obtained results indicated that nano CeO2 and SiO2 enhanced root growth and plan height of the P. polyphylla, respectively. This was due to the P. polyphylla absorbed SiO2 nano particles via roots and transferred them to the epidermis walls and vascular bundle of stem and leaves to protect as well as to induce growth of aboveground parts while the P. polyphylla also absorbed CeO2 nanoparticles and retained them in the epidermal roots to provide a medium culture accelerating the nutrient uptake of roots to significantly improve its growth. The simultaneously use of nano CeO2 and SiO2 greatly induced both root growth and plan height of the P. polyphylla. The extraction experiments suggested that significant amounts of gracillin, an important medicinal compound, accumulated in the P. polyphylla rhizome. Gracillin content in the rhizome of the CeO2 fertilized P. polyphylla was also greatly higher than that in the SiO2 fertilized P. polyphylla. Thus, the nano CeO2 not only promoted the development but also enhanced formation of gracillin in the rhizome of the P. polyphylla.

Antitumor effect of polyphyllin I (PPI) on colorectal cancer: Evidence from patient-derived organoids and Notch signaling suppression

Colorectal cancer (CRC) is a malignant tumor with a high incidence, ranking first among gastrointestinal malignancies. We investigated the impact of polyphyllin I (PPI), a natural compound found in Paris polyphylla, on CRC. PPI has been documented to exhibit anticancer activity against various tumors. This study aimed to assess the effects of PPI on colorectal cancer and explore its potential mechanisms. Our research demonstrated that PPI inhibited proliferation, promoted apoptosis, and induced G2 cell-cycle arrest in a dose-dependent manner. Additionally, our results indicated that PPI suppressed Notch signaling by downregulating the Notch1 receptor, its ligand Jagged1, and the downstream target Hes1 expression. Furthermore, we confirmed the antitumor effect of PPI on patient-derived organoids. In conclusion, our study indicates that PPI impedes the growth of colon cancer by suppressing the Notch signaling pathway.

Identification and validation of autophagy-related genes influenced by paris polyphylla in tongue cancer using network pharmacology

BackgroundTongue squamous cell carcinoma (TSCC) represents the most prevalent form of head and neck squamous cell carcinomas, comprising approximately one-third of all oral cancers. Paris polyphylla(PP) exhibit promising anti-tumor properties, yet their underlying mechanisms remain elusive. This study offers novel insights into the molecular mechanisms underlying TSCC treatment with PP and establishes a theoretical basis for their clinical application.MethodsEmploying transcriptomics and network pharmacology methodologies, we identified autophagy-related key genes associated with the effects of PP. These genes were subjected to KEGG and GO enrichment analyses to determine their related functions. In vitro, CAL-27 cells were treated with 10, 30, and 60 μg/ml of PP for 24 h to assess tumor cell proliferation, apoptosis, and autophagy-related markers.Key findings.Molecular docking of MAPK3 and PSEN1 with PP revealed stable hydrogen bond interactions, indicating the therapeutic potential of these saponins in TSCC through the autophagy pathway. In vitro experiments demonstrated significant inhibition of proliferative activity in tongue squamous carcinoma CAL-27 cells and promotion of tumor cell apoptosis by PP. Western blot analysis confirmed alterations in the expression of autophagy markers P62, LC3B, and Beclin1 following treatment, suggesting activation of the autophagy pathway.ConclusionsOur results suggest that PP inhibits tumor cells through the autophagy pathway, in which MAPK3 and PSEN1 play a role as potential functional molecules.

Occurrence of Maize Leaf Spot Disease Caused by Leptosphaerulina australis in Yunnan, China.

Maize (Zea mays) is vital as a staple food and livestock feed crop. Yunnan is one of the main maize-producing provinces in China (National Bureau of Statistics, 2022). While corn production in Yunnan is lower than the national average, the development of drought-tolerant varieties has contributed to improving productivity. In August 2021, a new leaf spot disease on maize was observed in Lancang, Yunnan (22°26'38.11"N to 22°48'38.68"N, 99°48'15.13"E to 99°59'20.03"E), causing serious damages to maize production with incidence up to 76.19 %. Initially, small light yellow lesions were seen scattered on diseased maize leaves, round or polygon, measuring 0.3 to 2.0 cm in diameter. In the intermediate phase, these lesions sank, ruptured, and turned white with dark brown borders. In severe cases, they merged into large irregular patches, reaching up to 10 cm, leading to complete leaf necrosis. Small black ascomata were seen on the lesions. Tissue sections reveal perithecium embedded in leaves, measuring 94~145 μm in diameter. Symptomatic tissues were sterilized in 1.5% NaClO for 60s, and washed twice withsterile purified water, then plated on potato dextrose agar (PDA) at 25℃, 90% relative humidity (RH), and a 12-hour light cycle. 6 isolates were obtained from 2 diseased maize cultivars. In 20 days, the colony reached the edge of the PDA plate, the center darkening from white, featuring white aerial mycelium on top and black on the reverse side. Brown ascomata, solitary or clustered, measured 80.1~176.7 × 55.57~138.9 μm. The ellipsoid to oblong ascospores were 17.9~39.7 × 10.9~14.1 μm, and the bitunicate, thick-walled asci were 90.1~133.3 × 26.6~33.5 μm. The genomic DNA was extracted using the Chelex-100 method (Möller et al. 1992). For molecular identification, the ITS, LSU, and β-tubulin (Tub2) genes were amplified using primer pairs ITS1/ITS4 (White et al. 1990), LR0R/LR5 (Vilgalys et al. 1990) and Btub2Fd/Btub4Rd (Woudenberg et al. 2009), respectively. Sequencing was performed by Sangon Biotech (Shanghai) Co., Ltd. The sequenced loci (GenBank accession nos.: LSU, OL687348-53; ITS, OL617009-10, and OL664058-61; Tub2, OL741678-83) of the isolates exhibited 100%/ 99%/ 100% similarities with L. australis genes: LSU, MH868885; ITS, KF381084; Tub2, GU237541, respectively. Using MEGA 11.0, phylogenetic trees were constructed using the maximum-likelihood algorithm on concatenated sequences of LSU, ITS, and Tub2 for isolates LCMB1 to 6. The isolates clustered with two L. australis strains with 100 % bootstrap support (1,000 replicates). The results were consistent with the Bayesian Inference tree. The pathogenicity test used strain LCMB4 on six healthy maize plants during the heading period under natural conditions. Three leaves pre-plant were wounded with sterile sandpaper and sprayed with conidial suspension (106 spores ml-1, diluted in sterilized water) in the greenhouse at 28℃, 90% RH, and a 12-hour light cycle, with sterilized distilled water used for control. Inoculated leaves developed symptoms consistent with the described after 10 days, while control leaves remained symptomless. The same pathogen was re-isolated from the infected leaves, fulfilling Koch's postulates. Previously, L. australis has been isolated from turfgrass (Mitkowski et al. 2004), Alfalfa (Zhang et al. 2021), soil (Li et al. 2018), and Paris polyphylla var. chinensis (Fu et al. 2019), but not from maize. This is the first report of L. australis causing leaf spot on maize globally.

Unveiling the hidden potential: Above-ground parts of Paris yunnanensis Franch. Is promise as an anti-acne therapeutic beyond traditional medicinal sites

The dried rhizomes of Paris yunnanensis Franch. have been extensively utilized in traditional Chinese medicine as hemostatic, antitumor, and antimicrobial agents. An examination of classical texts and renowned Chinese medical formulations showcased its efficacy in acne treatment. Presently, there is a significant scarcity of Paris resources. Consider directing attention towards the non-medicinal parts of Paris to mitigate the strain on medicinal resources within this realm. To address these resource limitations, this study investigated the bioactivity and pharmacodynamics of the above-ground parts of Paris (AGPP). A synergistic approach integrating network pharmacology, molecular docking (in silico validation), and animal experimentation (in vivo validation) was employed to elucidate the potential mechanisms underlying the efficacy of AGPP against acne vulgaris in this study. The active constituents in AGPP extracts were identified via UHPLC-Q-Orbitrap HRMS analysis, with their targets extracted for network pharmacological analysis. KEGG pathway analysis unveiled potential therapeutic mechanisms, validated through molecular docking and rat auricular acne model experiments. Comprehensive chemical characterization revealed fifty constituents, including steroidal saponins, flavonoids, amino acids, organic acids, phytohormones, phenolic acids, and alkaloids. Diosgenin, Quercetin, Kaempferol, Ecdysone, and α-linolenic acid were identified as main constituents with acne-treating potential. Core targets included SRC, MAPK3, and MAPK1, with key signaling pathways implicated. Histologically, AGPP mitigated acne-induced follicular dilatation and inflammation, inhibiting inflammatory cytokine production (IL-6, IL-1β, TNF-α). This study offers insight into AGPP's mechanism for acne treatment, laying groundwork for Paris development and drug discovery.

Study on Synthesis and Regulation of PPVI and PPVII in Paris polyphylla with UV.

Paris polyphylla Smith var. Chinensis (Franch.) Hara is a medicinal plant that belongs to the Liliaceae family. Its main components are parissaponins, which have excellent medicinal effects such as anti-inflammatory, anti-tumor, etc. Improving the quality of parissaponins through artificial directional regulation has emerged as a practice to meet medical demand and is a new research hotspot. In this paper, P. polyphylla plants were treated with UVA, UVB, and UVC, and the contents of PolyPhyllin VI (PPVI) and PolyPhyllin VII (PPVII), saponin synthase (squalene synthase, SS; cycloartenol synthase, CAS; cytochrome P450, CYP450; and glycosyl transferases, GT) activity, MDA, and the photosynthetic pigment indexes were measured and analyzed. The results showed that PPVII content increased by 32.43% with UVC treatment after 4 h (3.43 mg/g), but the PPVI and PPVII contents in the other groups decreased compared with CK (control group) and they did not return to the original level after 4 h. SS, CAS, CYP450, and GT synthases were activated in varying degrees via UV treatment and increased, respectively, by 22.93%, 10.83%, 20.15%, and 25.98%. Among them, GT, as the last of the synthetases, had a shorter response time to UVB (30 min) and UVC (15 min); the difference was sensible compared with CK. Moreover, UV had a stressing effect and promoted the rapid accumulation of MDA content (increased 17.66%, 34.53%, and 9.65%) and carotenoid (increased 7.58, 5.60, and 7.76 times) within 4 h compared to CK. UVB and UVC radiation visibly improved chlorophyll a content (42.56% and 35.45%), but UVA did not, and the change in chlorophyll b content showed no overt statistical difference. In addition, PPVI and PPVII were negatively correlated with SS, CAS, carotenoids, and MDA (p < 0.05) and positively correlated with CYP450, GT, and chlorophyll a (p < 0.05). This study provides a theoretical basis for using UV light to regulate secondary metabolism in P. polyphylla, which is of great value for production management.

Linking soil nutrients, microbial community composition, and enzyme activities to saponin content of Paris polyphylla after addition of biochar and organic fertiliser

The application of organic fertilisers and biochar has become widespread in agroforestry ecosystems to enhance the yield and quality of crops and medicinal plants. However, their specific impact on both the yield and quality of Paris polyphylla (P. polyphylla), along with the underlying mechanisms, remains unclear. In this study, we investigated the distinct effects of organic fertiliser (at concentrations of 5% and 10%) and biochar application (at levels of 2% and 4%) on P. polyphylla saponin content. This content is intricately regulated by available soil nutrients, enzyme activities, and microbial community compositions and activities. Our results clearly demonstrated a significant increase in the saponin content, including total saponin, polyphyllin I (PPI), polyphyllin II (PPII), polyphyllin VI (PPVI), and polyphyllin VII (PPVII), in P. polyphylla following the application of both biochar and organic fertiliser. Moreover, in comparison to the control group, the addition of biochar and organic fertiliser led to a considerable rise in the activity of glycosyltransferase enzyme (GTS) and cycloartenol synthase (CAS) in P. polyphylla. Additionally, it increased soil available potassium (AK) and soil organic matter (SOM) concentration, along with the activity of urease, acid phosphatase, and catalase, although biochar amendment resulted in a decrease in nitrate nitrogen (NO3−-N) concentration. Crucially, our findings revealed a positive correlation between total saponin content and the activity of CAS in P. polyphylla, soil AK, SOM concentration, and the activities of urease, acid phosphatase, and catalase. Conversely, there was a negative correlation with NO3−-N content. Furthermore, the application of organic fertiliser and biochar significantly influenced microbial community structures and specific microbial taxa. Notably, total saponin content exhibited a positive relationship with the relative abundances of Dehalococcoidia, Saccharomycetes, and Agaricomycetes taxa while showing a negative correlation with the abundance of Verrucomicrobiae. In conclusion, the observed increase in saponin content can be attributed to the modulation of specific microbial taxa in soils, as well as alterations in soil nutrients and enzyme activities resulting from the application of biochar and organic fertiliser. This study identifies a potential mechanism for enhancing saponin content in the artificial cultivation of P. polyphylla.

Optimization of ultrasound-assisted extraction of two saponins from Paris polyphylla var. yunnanensis leaves using response surface methodology

Paris polyphylla var. yunnanensis is one of the famous Chinese herbs, in which two saponins (polyphyllin II and polyphyllin VII) have anticancer effects. The endangerment of Paris polyphylla var. yunnanensis, makes the study of optimizing the extraction of polyphyllin II and polyphyllin VII from the leaves of Paris polyphylla var. yunnanensis more important. The study established and optimized the process of ultrasound-assisted extraction for polyphyllin II and polyphyllin VII using the Box Behnken Design method of response surface methodology. The results showed that the optimum extraction conditions for polyphyllin II and polyphyllin VII are ethanol concentration of 73 and 70%, extraction temperature of 43 and 50°C, and number of extraction 3, respectively. Under the above conditions, the contents of polyphyllin II and polyphyllin VII were measured to be 6.427 and 19.015 mg/g (DW). The results showed that the experimental model fitted well, and the response surface methodology (RSM) was feasible to optimize the extraction process of polyphyllin II and polyphyllin VII from Paris polyphylla var. yunnanensis leaves. This method provides an effective approach for the comprehensive development and utilization of non-medicinal parts of Paris polyphylla var. yunnanensis.

Integrated Analysis of Transcriptome and Metabolome Reveal the Enhancement of Methyl Dihydrojasmonate on Physiological Indicators and Polyphyllins Biosynthesis in Paris polyphylla var. yunnanensis

Integrated Analysis of Transcriptome and Metabolome Reveal the Enhancement of Methyl Dihydrojasmonate on Physiological Indicators and Polyphyllins Biosynthesis in Paris polyphylla var. yunnanensis

Two new saponins from the rhizomes of Paris yunnanensis Franch

The phytochemical investigation on the rhizomes of Paris yunnanensis Franch. resulted in the discovery and characterisation of six compounds, including two new saponins named parisyunnanosides M-N (1–2), and four known ones (3–6). The structures of isolated compounds were determined by spectroscopic data analysis and chemical methods. Compound 2 is a pregnane-type saponin with a special α,β-unsaturated carboxylic acid moiety at C-17, which is first discovered in genus Paris. The anti-inflammatory activity of the isolated compounds was assessed in vitro. The results demonstrated that compounds 3 and 4 could significantly inhibit the production of NO which was induced by LPS in RAW 264.7 cells with IC50 values of 0.67 ± 0.17 μM and 0.85 ± 0.12 μM, respectively.

Colletotrichum paridis sp. nov., a novel endophytic species on Paris polyphylla var. chinensis

The genus Paris is a crucial medicinal herb, widely cultivated in East Asia and Europe. In 2017 a novel Colletotrichum species was isolated from leaf lesions with grey mould of Paris polyphylla var. chinensis in Sichuan Province of China. Through single-spore technique, four isolates were obtained for characterization. Phylogeny inferred from multi-locus sequences of ITS, gapdh, act, tub2, chs-1, and his3, showed that these isolates represented a novel sister species to C. jinshuiense within the C. dematium complex. Morphologically, this new species produces larger conidia than those of C. jinshuiense and cultures on PDA show a noticeable pink ring. Tests showed that this species had no pathogenicity to the host plants. Based on phylogenetic, morphological, and pathogenicity data, we propose the name C. paridis sp. nov. for this new endophyte from P. polyphylla in China.

Characterization of a glycosyltransferase from Paris polyphylla for application in biosynthesis of rare ophiopogonins and ginsenosides

Saponins are bioactive components of many medicinal plants, possessing complicated chemical structures and extensive pharmacological activities, but the production of high-value saponins remains challenging. In this study, a 6′-O-glucosyltransferase PpUGT7 (PpUGT91AH7) was functionally characterized from Paris polyphylla Smith var. yunnanensis (Franch.) Hand. -Mazz., which can transfer a glucosyl group to the C-6′ position of diosgenin-3-O-rhamnosyl-(1 → 2)-glucoside (1), pennogenin-3-O-rhamnosyl-(1 → 2)-glucoside (2), and diosgenin-3-O-glucoside (5). The KM and Kcat values of PpUGT7 towards the substrate 2 were 8.4 μM and 2 × 10−3 s−1, respectively. Through molecular docking and site-directed mutagenesis, eight residues were identified to interact with the sugar acceptor 2 and be crucial for enzyme activity. Moreover, four rare ophiopogonins and ginsenosides were obtained by combinatorial biosynthesis, including an undescribed compound ruscogenin-3-O-glucosyl-(1 → 6)-glucoside (10). Firstly, two monoglycosides 9 and 11 were generated using a known sterol 3-O-β-glucosyltransferase PpUGT80A40 with ruscogenin (7) and 20(S)-protopanaxadiol (8) as substrates, which were further glycosylated to the corresponding diglycosides 10 and 12 under the catalysis of PpUGT7. In addition, compounds 7–11 were found to show inhibitory effects on the secretion of TNF-α and IL-6 in macrophages RAW264.7. The findings provide valuable insights into the enzymatic glycosylation processes in the biosynthesis of bioactive saponins in P. polyphylla var. yunnanensis, and also serve as a reference for utilizing UDP-glycosyltransferases to construct high-value or rare saponins for development of new therapeutic agents.

Validating the feasibility of the method of ensemble learning combined with FT-MIR for the discrimination of wild Paris polyphylla var. Yunnanensis from different geographical sources

As a widely used medicinal plant with high economic value, Paris polyphylla var. yunnanensis (PPY), is mainly distributed in south-western China. Since different growing environments have a strong influence on the PPY quality, efficient means are needed to discriminate its origin source. The traditional machine learning model uses a single learner to classify and discriminate the samples, but it is difficult to apply its advantages to play for complex data samples. Ensemble learning can integrate the advantages of multiple learners to improve the model performance. In this experiment, Partial Least Squares Discriminant Analysis (PLS-DA), Random Subspace Method (RSM), Random Forest (RF), and Support Vector Machine (SVM) models were used to discriminate wild PPY from different origins, and then verify the feasibility of ensemble learning in discriminating them from different geographical sources. At the same time, the results of preprocessing the spectral data with multiplicative scattering correction (MSC), standard normal variate (SNV), and derivative operations (first, second and third derivatives) and their combination methods were compared. The results show that the combined preprocessing greatly improves the model performance. The RSM has the most stable performance with an accuracy of more than 85%. Indicating that the ensemble learning combined with Fourier transform mid-infrared spectroscopy (FT-MIR) has the feasibility of discriminating the wild PPY of different geographical origins.

Study on Anti-fatigue Effects and Mechanisms of Polysaccharide from Paris polyphylla.

The objectives of this study were to investigate the anti-fatigue effects of Paris polyphylla polysaccharide component 1 (PPPm-1) and explore its mechanisms. A mouse model of exercise-induced fatigue was established by weight-bearing swimming to observe the effects of different concentrations of PPPm-1 on weight-bearing swimming time. The anti-fatigue effect of PPPm-1 was determined by the effects of contraction amplitude, contraction rate, and diastolic rate of the frog gastrocnemius muscle in vivo before and after infiltration with 5 mg/mL PPPm-1. The effects of PPPm-1 on the contents of blood lactate, serum urea nitrogen, hepatic glycogen, muscle glycogen in the exercise fatigue model of mice, and acetylcholine (ACh) content and acetylcholinesterase (AChE) activity at the junction of the frog sciatic nerve-gastrocnemius under normal physiological, and Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of the frog gastrocnemius were determined by enzyme-linked immunosorbent assay (ELISA), to investigate the anti-fatigue mechanisms of PPPm-1. The results showed that PPPm-1 could significantly prolong the weight-bearing swimming time in mice (P < 0.01), decrease the contents of blood lactate and serum urea nitrogen, increase the contents of the hepatic glycogen and muscle glycogen of mice after exercise fatigue compared with those of the control group, and there was extremely significant difference in most indicators (P < 0.01). The 5mg/mL of PPPm-1 could significantly promote the contraction amplitude, contraction rate, and relaxation rate of the gastrocnemius muscle in the frogs, and the content of ACh at the junction of the frog sciatic nerve-gastrocnemius (P < 0.01), but it had obvious inhibitory effetc on the activity of AChE at the junction of the frog sciatic nerve-gastrocnemius (P < 0.01). PPPm-1 could increase the Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of gastrocnemius in the frogs (for Ca2+-Mg2+-ATPase, P < 0.01). The above results suggested that the PPPm-1 had a good anti-fatigue effect, and its main mechanisms were related to improving endurance and glycogen reserve, reducing glycogen consumption, lactate and serum urea nitrogen accumulation, and promoting Ca2+ influx.