Polyphylla Smith Var Research Articles

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.

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.

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.

Comparative genomic analysis and proposal of Clostridium yunnanense sp. nov., Clostridium rhizosphaerae sp. nov., and Clostridium paridis sp. nov., three novel Clostridium sensu stricto endophytes with diverse capabilities of acetic acid and ethanol production

ObjectivesGenus Clostridium sensu stricto is generally regarded as the true Clostridium genus, which includes important human and animal pathogens and industrially relevant microorganisms. Besides, it is also a prominent member of plant-associated endophytes. However, our knowledge of endophytic Clostridium is limited. MethodsIn this study, the endophytes were isolated under anaerobic condition from the roots of Paris polyphylla Smith var. yunnanensis. Subsequently, a polyphasic taxonomic approach was used to clarify their taxonomic positions. The fermentation products were measured in the isolates with HPLC analysis. Comparative genomics was performed on these new strains and other relatives. ResultsIn total, nine endophytic strains belonging to the genus Clostridium sensu stricto were isolated, and three of them were identified as new species. Seven of nine strains could produce acetate, propionate, and butyrate. Only two strains could produce ethanol, although genomics analysis suggested that only two of them were without genes for solventogenesis. Different from the endophytic strains, the phylogenetically closely related non-endophytic strains showed significant enrichment effects on some metabolic pathways involving environmental information processing, carbohydrate, and amino acid metabolisms, etc. It suggests that the genomes of these endophytic strains had undergone subtle changes associated with environmental adaptations. ConclusionConsequently, strains YIM B02505T, YIM B02515T, and YIM B02565T are proposed to represent a new species of the genus Clostridium sensu stricto, for which the names Clostridium yunnanense sp. nov., Clostridium rhizosphaerae sp. nov., and Clostridium paridis sp. nov. are suggested.

Azospirillum Endophyticum sp. nov., an Endophyte of Paris Polyphylla Smith var. Yunnanensis.

A Gram-negative, facultative anaerobic bacterial strain, designated YIM B02556T, was isolated from the root of Paris polyphylla Smith var. yunnanensis collected from Yunnan Province, southwest China. By using a polyphasic approach, its taxonomic position was investigated. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM B02556T belonged to the genus Azospirillum and the 16S rRNA gene sequence similarity values of strain YIM B02556T to the type strains of members of this genus ranged from 94.9 to 98.3%. Overall genome relatedness index (OGRI) analysis estimated based on average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) between YIM B02556T and other Azospirillum species type strains were <90.8% and <37.8%, lower than the limit of species circumscription. Cells of the strain were characterized as oxidase- and catalase-positive, with motility provided by flagella. The growth conditions of the strain were found to occur at 20-40°C (optimum, 35°C), and pH 6.0-9.5 (optimum, pH 7.5). Strain YIM B02556T can tolerate 2% NaCl concentration. Strain YIM B02556T contained Q-10 as the major ubiquinone. The major fatty acids were C18:1 ω7c and summed feature three (C16:1 ω7c and/or C16:1 ω6c). The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. Based on polyphasic analysis, strain YIM B02556T could be differentiated genotypically and phenotypically from recognized species of the genus Azospirillum. Therefore, the isolate represents a novel species, for which the name Azospirillum endophyticum is proposed. The type strain is YIM B02556T (=JCM 34631T=CGMCC 1.18654T).

Synergy Mechanisms of Rhizoma Paridis Saponins on Non-small Cell Lung Cancer: Segmented Solid Phase Extraction, Bioactivity Screening, and Network Pharmacology.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. Rhizoma paridis saponins (RPS), the main bioactive ingredients of Paris polyphylla Smith var. yunnanensis (PPY), have been proved to have remarkable effects on NSCLC cell lines. However, the multi-component synergistic effects and mechanisms of RPS on NSCLC have not been elucidated. To decipher the multi-RPS synergistic effects and mechanisms against NSCLC based on network pharmacology combined with segmented solid-phase extraction (SPE) and bioactivity screening method. Firstly, segmented SPE and cytotoxicity assays were performed to screen the RPS-enrichment fraction of PPY, and the steroidal saponins in it were identified by LC-MS/MS. Then, a network pharmacology analysis was performed to predict the potential therapeutic targets of RPS on NSCLC. Finally, viable cell counting tests and RT-qPCR were utilized to verify the synergistic effects and mechanisms of RPS. 48 potentially active compounds were identified from the 30% MeOH/EtOAc fraction of PPY (30% M/E PPY). The results of the network pharmacology analysis indicated that RPS exerted joint effects by regulating six key targets in the PI3K-AKT signaling pathway. In vitro experiments showed that due to the synergistic effects, 30% M/E PPY at 13.90 μg/mL could exert a stronger inhibitory activity on A549 cells by reducing the overexpression of six hub genes compared with the parallel control groups. This research elaborates on the multi-RPS synergy mechanisms against NSCLC and provides a way to develop new combination medicines for NSCLC.

Paris panxiensis, a new combination of Trilliaceae from Sichuan, China

Abstract This article describes and illustrates Paris panxiensis (J. L. Liu) Y. Yuan &amp; J. L. Liu from a morphological perspective, based on field survey, indoor potting and garden cultivation, and observations gleaned from these activities. This variety is closely related to P. delavayi Franch, P. daliensis H. Li et V. G. Soukup, P. lihengiana G. W. Hu &amp; Q. F. Wang and P. polyphylla Smith var. stenophylla Franch, but differs from these in having the following characteristics: flower parts with a basic number of 4–8, petals far shorter than sepals, linear-virgate, bottom linear, gradually thickening upward, erect, never extend or curved, apex rounded or obtuse, and connectivum projecting.

A new variety from Sichuan, China—Paris polyphylla var. nullopetalina (Trilliaceae)

Abstract This paper succinctly describes and then illustrates the species Paris polyphylla Smith var. nullopetalina J. L. Liu et Y. Yuan, which is a new variety of Trilliaceae found in Sichuan Province in Southwestern China. It is closely related to Paris polyphylla Smith var. panxiensis J. L. Liu, Paris polyphylla Smith var. stenophylla Franch and Paris polyphylla Smith var. minora S. F. Wang. The distinguishing features that can be used to identify this variety are as follows: the leaf blade is narrowly long, fasciate and crisped-margined, but differs in having leaves that are sessile, with petals absent, ovary tetragonal, and stigma erect and never curved.

Two new polyhydroxylated steroidal glycosides from Paris polyphylla var. yunnanensis

Chemical investigation of Paris polyphylla smith var. yunnanensis afforded two new polyhydroxylated steroidal glycosides, named Parisyunnanosides K and L ( 1 – 2 ), together with nine known ones. The chemical structures of the new compounds were elucidated by 1D, 2D NMR and HR-ESI-MS techniques, together with chemical methods. Parisyunnanosides K and L ( 1 – 2 ) are rare C 27 steroidal glycosides with two double bonds located at C-5, 6 and C-25, 26 of the aglycone, respectively. The cytotoxic activities of the isolated compounds were evaluated against Caco-2 cells by CCK-8 assay. Among them, compounds 5 – 11 exhibited potent cytotoxic activity with IC 50 values ranging from 1.10 to 14.14 μM compared with the positive control oxaliplatin (1.38 μM). Eleven steroidal saponins, including two previously unreported, were isolated from the rhizomes of P. polyphylla var. yunnanensis. The isolates 1 – 11 were evaluated for their cytotoxic activity in Caco-2 cells by using CCK-8 assay. • Two previously undescribed steroidal glycosides were characterized from Paris polyphylla . • The new compounds are rare C 27 steroidal glycosides with two double bonds of the aglycone. • Cytotoxic activities of the isolated compounds were tested on Caco-2 cancer cells.

Paenibacillus terrae PY8 controls Botrytis cinerea and modifies the endophytic microbial community of the medicinal plant, Paris polyphylla

Huachonglou (Paris polyphylla Smith var. chinensis (Franch.) Hara) is an important traditional Chinese herbal medicine. Botrytis cinerea is a plant pathogen that causes significant economic losses of P. polyphylla. Presently, chemical fungicides are the primary means of effective control of gray mold caused by B. cinerea. As a safer and greener control method, biocontrol agents (BCAs) for gray mold of P. polyphylla are very important. In the present study, a biocontrol agent, Paenibacillus terrae PY8, was isolated from the endophytic microbial community of leaves of the medicinal plant, Paris polyphylla. PY8 significantly reduced gray mold infections by Botrytis cinerea and additionally increased P. polyphylla yield and the level of polyphyllin Ⅰ and Ⅱ of P. polyphylla detected in rhizomes. An analysis of the endophytic microbial community of leaves of P. polyphylla indicated that the foliar application of P. terrae PY8 favorably altered the composition and structure of the endophytic community in P. polyphylla by decreasing the total abundance of potential plant pathogenic fungi. The effect of biocontrol agents on the endophytic microbiome may represent a new criterion for evaluating biocontrol agents in the future. Results of the present study provide theoretical support for the field application of PY8 on P. polyphylla for the control of gray mold.

Neobacillus paridis sp. nov., an endophyte of Paris polyphylla Smith var. yunnanensis.

A novel endophytic strain, designated YIM B02564T, was isolated from the root of Paris polyphylla Smith var. yunnanensis obtained from Yunnan Province, southwest China. By using a polyphasic approach, cells of the strain were characterized as facultative anaerobic, Gram-positive and rod-shaped. The growth conditions of the strain were found to occur at 20-55°C (optimum, 30°C), pH 6.0-9.0 (optimum, pH 7.0). Strain YIM B02564T can tolerate 2% NaCl concentration. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM B02564T belonged to the genus Neobacillus and the 16S rRNA gene sequence similarity values of strain YIM B02564T to the type strains of members of this genus ranged from 95.6 to 97.8%. The DNA G+C content of strain YIM B02564T calculated from the whole genome sequence was 41.6mol%. Values of the ANI and the dDDH between strain YIM B02564T and its closely related Neobacillus species were below 77.9% and 21.5%. Strain YIM B02564T contained MK-7 as the major menaquinone, iso-C15:0 and anteiso-C15:0 as the major fatty acids. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid and four unidentified lipids. It contained meso-diaminopimelic acid in the cell-wall peptidoglycan. On the basis of polyphasic analysis, strain YIM B02564T could be differentiated genotypically and phenotypically from recognized species of the genus Neobacillus. The isolate therefore represents a novel species, for which the name Neobacillus paridis is proposed. The type strain is YIM B02564T (= JCM 34668T = CGMCC 1.18655T).

Janibacter endophyticus sp. nov., an Endophytic Actinobacterium Isolated from the Root of Paris polyphylla Smith var. Yunnanensis.

A novel endophytic actinobacterium, designated as strain YIM B02568T, was isolated from the root of Paris polyphylla Smith var. Yunnanensis obtained from Yunnan Province, southwest China. Strain YIM B02568T was characterized using a polyphasic approach. Phylogenetic analysis indicated that this isolate belonged to the genus Janibacter. The 16S rRNA gene sequence similarity values of strain YIM B02568T to the type strains of members of this genus ranged from 95.8 to 98.6%. However, overall genome relatedness indices were significantly lower than the widely accepted species-defined threshold. The cell wall of strain YIM B02568T contained meso-diaminopimelic acid. The major menaquinone was MK-8(H4). The main polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylinositol. The major cellular fatty acids were comprised of iso-C16:0 and C18:1 ω9c. The DNA G + C content was 71.6mol%. Based on the data from the polyphasic studies, we propose that strain YIM B02568T represents a novel species within the genus Janibacter, Janibacter endophyticus sp. nov. The type strain is YIM B02568T (= JCM 34639T = CGMCC 1.18658T).

Chryseobacterium paridis sp. nov., an endophytic bacterial species isolated from the root of Paris polyphylla Smith var. yunnanensis.

A Gram-negative, yellow-pigmented, rod-shaped bacterial strain YIM B02567T was isolated from the root of Paris polyphylla Smith var. yunnanensis in China. Strain YIM B02567T grew optimally at 25-30°C and at pH 7.0 in the absence of NaCl on nutrient agar. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain YIM B02567T belong to the genus Chryseobacterium, and was closely related to Chryseobacterium piperi CTMT and Chryseobacterium soli DSM 19298T. Whole genome sequencing indicated that the genome size was 4,774,612bp and with a G + C content of 34.5mol%. Values of the ANI and the dDDH between strain YIM B02567T and its closely related Chryseobacterium species were below 81.72% and 24.7%. Strain YIM B02567T contained menaquinone-6 as the sole isoprenoid quinone, anteiso-C15:0, iso-C17:1 ω9c and iso-C17:0 3-OH as major fatty acids and phosphatidylethanolamine as major polar lipid. Based on the polyphasic analyses, strain YIM B02567T could be differentiated genotypically and phenotypically from recognized species of the genus Chryseobacterium. The isolate, therefore, represents a novel species, for which the name Chryseobacterium paridis sp. nov. is proposed. The type strain is YIM B02567T (= CGMCC 1.18657T).

Cloning and Functional Characterization of a Squalene Synthase from Paris polyphylla var. yunnanensis.

Paris polyphylla Smith var. yunnanensis (Franch.) Hand. - Mazz. is a precious traditional Chinese medicine, and steroidal saponins are its major bioactive constituents possessing extensive biological activities. Squalene synthase (SQS) catalyzes the first dedicated step converting two molecular of farnesyl diphosphate (FDP) into squalene, a key intermediate in the biosynthetic pathway of steroidal saponins. In this study, a squalene synthase gene (PpSQS1) was cloned and functionally characterized from P. polyphylla var. yunnanensis, representing the first identified SQS from the genus Paris. The open reading frame of PpSQS1 is 1239 bp, which encodes a protein of 412 amino acids showing high similarity to those of other plant SQSs. Expression of PpSQS1 in Escherichia coli resulted in production of soluble recombinant proteins. Gas chromatography-mass spectrometry analysis showed that the purified recombinant PpSQS1 protein could produce squalene using FDP as a substrate in the in vitro enzymatic assay. qRT-PCR analysis indicated that PpSQS1 was highly expressed in rhizomes, consistent with the dominant accumulation of steroidal saponins there, suggesting that PpSQS1 is likely involved in the biosynthesis of steroidal saponins in the plant. The findings lay a foundation for further investigation on the biosynthesis and regulation of steroidal saponins, and also provide an alternative gene for manipulation of steroid production using synthetic biology.

Molecular-networking-guided discovery of species-specific markers for discriminating five medicinal Paris herbs

BackgroundParidis Rhizoma (PR) is a famous traditional herbal medicine. Apart from two officially recorded species, viz. Paris polyphylla Smith var. yunnanensis (Franch.) Hand. - Mazz. (PPY) and P. polyphylla Smith var. chinensis (Franch.) Hara (PPC), there are still many other species used as folk medicine. It is necessary to understand the metabolic differences among Paris species. PurposeTo establish a strategy that can discover species-specific steroidal saponin markers to distinguish closely-related Paris herbs for quality and safety control. MethodsA new strategy of molecular-networking-guided discovery of species-specific markers was proposed. Firstly, the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was applied to obtain the MS and MS/MS data of all samples. Then, molecular networking (MN) was created using MS/MS data to prescreen the steroidal saponins for subsequent analysis. Next, the principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA) models were established to discover potential markers. Finally, the verification, identification and distribution of chemical markers were performed. ResultsA total of 126 steroidal saponins were screened out from five species using MN. Five species were classified successfully by OPLS-DA model, and 18 species-specific markers were discovered combining the variable importance in the projection (VIP) value, P value (one-way ANOVA) and their relative abundance. These markers could predict the species of Paris herbs correctly. ConclusionThese results revealed that this new strategy could be an efficient way for chemical discrimination of medicinal herbs with close genetic relationship.

Paenibacillus paridis sp. nov., an endophytic bacterial species isolated from the root of Paris polyphylla Smith var. yunnanensis.

A Gram-reaction-positive, endospore-forming and rod-shaped bacterial strain, designated py1325T, was isolated from the root of Paris polyphylla Smith var. yunnanensis collected from Yunnan Province, PR China, and subjected to a polyphasic taxonomic characterization. It grew optimally with 0-1 % NaCl (w/v), at pH 7 and at 30 °C. The major respiratory quinone was MK-7 and the diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major cellular fatty acid was anteiso-C15 : 0. The polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, two unidentified aminophospholipids, two unidentified phospholipids and two unidentified lipids. The results of 16S rRNA gene sequence analysis revealed the highest levels of sequence similarity with respect to Paenibacillus luteus R-3T (99.0 %), Paenibacillus sinopodophylli CCTCC AB 2016047T (97.9 %), Paenibacillus castaneae DSM 19417T (97.5 %) and Paenibacillus endophyticus LMG 27297T (97.2 %). The digital DNA-DNA hybridization and average nucleotide identity values between py1325T and these species ranged 20.6-53.3 % and 79.9-93.6 %. The G+C content of the genomic DNA was 47.7 mol%. According to the phylogenetic, phenotypic and chemotaxonomic evidence, strain py1325T clearly represents a novel species of the genus Paenibacillus, for which the name Paenibacillus paridis sp. nov. is proposed. The type strain is py1325T (=CCTCC AB 2015220T=LMG 29068T).

Determination of Inorganic Elements in the Rhizome of Paris polyphylla Smith Var. chinensis (Franch.) Hara by Using Inductively Coupled Plasma Mass Spectrometry.

The objective of this study was to investigate the concentrations of inorganic elements in the rhizome of Paris polyphylla Smith var. chinensis (Franch.) Hara of different planting years and cultivation conditions. Twenty-five inorganic elements including Al, As, B, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Na, Ni, P, Pb, Se, Sr, Ti, V, and Zn in the rhizome were determined by using inductively coupled plasma mass spectrometry (ICP-MS). The analytical method was validated by measuring several parameters including linearity, correlation coefficient, limit of detection (LOD), limit of quantification (LOQ), and recovery. The linear working ranges were three, 0–300 μg/L, 0–500 μg/L, and 0–1000 μg/L, and the correlation coefficients (r) values were higher than 0.998. LOD varied from 0.001 μg/L (Be) to 11.957 μg/L (P), and LOQ varied from 0.003 μg/L (Be) to 35.870 μg/L (P). The recoveries spanned from 95.2 (Co) to 105.3% (Pb). Validation parameters showed the possibility of using whole of the sample preparation procedures used in this study. Based on the determined results, it is indicated that the toxic elements As, Cd, and Pb had no ingestion risk. The planting years and cultivation conditions had significant effects on the concentrations of inorganic elements of P. polyphylla var. chinensis. The concentrations of inorganic elements in cultivated samples were different from those in wild samples. The results can provide useful theoretical basis for the quality control and rational use of P. polyphylla var. chinensis.

EFFECT OF PROVENANCE AND WATER STRESS ON BIOMASS AND POLYPHYLLIN CONTENT IN THE MEDICINAL PLANT Paris polyphylla Smith var. yunnanensis

Water stress and provenance could affect the secondary metabolites synthesis and accumulation in herbs. Thus, this study explored the effect of soil water moisture and provenance on the growth of Paris polyphylla Smith var. yunnanensis (PPY). Three provenances (Jinping, Luquan and Weixi in Yunnan, China) of PPY samples were grown in different soil water moisture conditions [0.80, 0.70 and 0.50 field capacity (FC)] during Dec. 2015 to Sep. 2017. Results showed that the highest biomass weight was presented in 0.70 FC for Luquan and Weixi samples. Biomass weight for Jinping provenance presented a decreasing tendency with the decreased soil water moisture and the highest biomass were shown in 0.80 FC. However, quantitative analysis revealed that the total content of polyphyllin increased with decreasing the soil water moisture for Jinping and Weixi samples. The highest total content of polyphyllin in rhizome was inclined to show in Jinping samples, while the stem and leaf tissues were shown in Weixi samples. Additionally, results of ANOVA combined with PCA indicated that the difference among these three provenances were significant. Correlation analysis results revealed that 0.50 FC induced the competitive relationship occurrence for polyphyllin distribution. Thus, 0.70 FC was the most suitable soil-water condition for PPY growth. Besides, provenance collected from Jinping could consider as a good quality germplasm. Consequently, this study might provide a preliminary foundation for irrigation project formulated and provenance screened for PPY cultivation.

Structural characterisation and discrimination of the aerial parts of Paris polyphylla var. yunnanensis and Paris polyphylla var. chinensis by UHPLC-QTOF-MS coupled with multivariate data analysis.

As sources of Rhizoma Paridis are facing shortages, utilising the aerial parts of Paris polyphylla has emerged as a promising additional source. However, the components in the aerial parts still need to be explored, and it is difficult to distinguish the aerial parts of P. polyphylla Smith var. yunnanensis (PPY) and P. polyphylla var. chinensis (PPC), two varieties of P. polyphylla. This study aimed to establish a comprehensive platform to characterise steroid saponins from the aerial parts of PPY and PPC and to discriminate these two varieties. A dereplication approach and ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) analysis were used for the characterisation of steroidal saponins in the aerial parts of PPY and PPC. Multivariate statistical analysis was performed to differentiate these two varieties and screen discriminant variables. In addition, a genetic algorithm-optimised for support vector machines (GA-SVM) model was developed to predict P. polyphylla samples. The distribution of steroidal saponins in PPY and PPC was visualised by a heatmap. A total of 102 compounds were characterised from the aerial parts of PPY and PPC by dereplication. A clear separation of PPY and PPC was achieved, and 35 saponins were screened as marker compounds. The established GA-SVM model showed excellent prediction performance with a prediction accuracy of 100%. Many steroid saponins that have been reported in Rhizoma Paridis also exist in the aerial parts of P. polyphylla. Samples from the aerial parts of PPY and PPC could be discriminated using our platform.

Traceability the provenience of cultivated Paris polyphylla Smith var. yunnanensis using ATR-FTIR spectroscopy combined with chemometrics

The conventional procedures, based on attenuated total reflectance-Fourier transform infrared spectrometry (ATR-FTIR), have been developed for the origins traceability of cultivated Paris polyphylla Smith var. yunnanensis (PPY) samples with the help of partial least square discriminant analysis (PLS-DA) and random forest. In this study, a set of 219 batch cultivated PPY samples, containing the cultivation years of 5, 6 and 7, and covering the municipal districts of Chuxiong, Dali, Honghe, Lijiang and Yuxi in Yunnan Province, China, were used to build the discrimination models. Firstly, a visualized analysis was carried out by t-distributed stochastic neighbor embedding (t-SNE) to reduce each data point in a two-dimensional map and make a knowledge of the sample distribution tendency. Secondly, the single spectra data sets of Paridis rhizome and leaf tissues, and the combination of these two data sets with variable selection (mid-level data fusion strategy), were used to establish PLS-DA and random forest models, and parallelly compared the model performance. Results demonstrated that the discrimination ability of PLS-DA preceded the random forest model, and the classification performance was remarkably improved after mid-level data fusion. These results verified each other by 5-, 6- and 7-year old Paridis samples and indicated that the model performance established in the present study was reliable. Besides, five agronomic characters, including the plant height, dry weight of rhizome and leaf tissues, and the allocation of rhizome and leaf were determined and analyzed, results of which indicated that the dry weight and their allocation was significantly different among various origins and fluctuated with the cultivation years. This study was using a comprehensive and green analytical method to discriminate the cultivated Paridis according to their provenances, which was simultaneously benefited for the appropriate cultivation areas selection based on the dry weight of rhizome tissues.