The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent r...The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent regulation of PPC seed dormancy and germination,we investigated the dynamic changes in PPC seed transcript levels under warm and cold temperature stratifications(WS and CS,respectively)by time-resolved RNA sequencing,focusing on genes related to hormone metabolism and signaling and cell wall remodeling(CWRM)and encoding transcription factors/regulators(TFs/TRs).A total of 48765 and 47836 differentially expressed genes(DEGs)were associated with WS and CS,respectively.Of these,17581 and 16652 DEGs,respectively,unique to WS and CS,and 5386 were common to both temperature stratifications across nine analyzed growth stages.The expression of hormone metabolism and signaling,TF/TR,and CWRM genes were closely associated with temperature.More genes related to gibberellin(GA),cytokinin,auxin,and brassinosteroid biosynthetic were upregulated in WS as compared to CS seeds,while genes related to dormancy release and germination were downregulated in WS seeds.However,only GA and abscisic acid levels were altered in PPC seeds breaking morphophysiological dormancy(MPD).Overall,37 TF and five TR families were upregulated whereas 24 TF and 16 TR families were downregulated in WS as compared to CS seeds.Most CWRM families were highly expressed under WS as compared to CS,suggesting that they promote endosperm weakening and embryo growth of WS seeds and facilitate MPD release and germination.A coexpression analysis revealed positive correlations between TFs/TRs and DEGs involved in hormone metabolism and signaling and CWRM.These results provided a basis for investigating the interaction between the endosperm and underdeveloped embryo in the regulation of PPC seed germination and seedling emergence.展开更多
Arbuscular mycorrhizal fungi(AMF)are important members of the plant microbiome and affect the uptake and transfer of mineral elements by forming a symbiotic relationship with plant roots.Nitrogen(N),as an important mi...Arbuscular mycorrhizal fungi(AMF)are important members of the plant microbiome and affect the uptake and transfer of mineral elements by forming a symbiotic relationship with plant roots.Nitrogen(N),as an important mineral element,can directly affect plant growth and development at different N levels.It has been confirmed that inoculation with AMF can improve the efficiency of N utilization by plants.However,there are still fewer reports on the dynamic relationship between arbuscular mycorrhizal and plant secondary metabolites at different nitrogen levels.In this experiment,the physiological indexes and genes related to saponin synthesis were determined by applying different concentration gradients of nitrogen to the medicinal plant P.polyphylla var.yunnanensis infested by AMF as the test material.It was found that nitrogen addition increased the biomass,chlorophyll content,and nutrient content of above-and below-ground plant parts and increased the content of saponin content of P.polyphylla var.yunnanensis to some extent,but AMF inoculation increased the saponin content of P.polyphylla var.yunnanensis more significantly.AMF inoculation also promoted the expression of genes related to the saponin synthesis pathway,including 3-hydroxy-3-methylglutaryl coenzyme A synthase(HMGS),squalene epoxidase 1(SE1),and cycloartenol synthase(CAS),which is in according with the accumulation of saponin in plants.It also may increase the saponin content of AMF plants by altering the expression of P450s and UGTs related to saponin synthesis.展开更多
[Objective] This study aimed to isolate the endophytic fungi of Paris polyphylla var. yunnanensis and investigate their effects on the embryo development of P. polyphylla var. yunnanensis seeds. [Method] The endophyti...[Objective] This study aimed to isolate the endophytic fungi of Paris polyphylla var. yunnanensis and investigate their effects on the embryo development of P. polyphylla var. yunnanensis seeds. [Method] The endophytic fungi of P. polyphylla were isolated and identified morphologically, and their effects on the embryo development of P. polyphylla var. yunnanensis seeds were studied by using paraffin sectioning and microphotography. [Result] Nine endophytic fungi, i.e. P. polyphylla var. yunnanensis endophytic fungi PPYEF-1, PPYEF-2, PPYEF-3, PPYEF-4, PPYEF-5, PPYEF-6, PPYEF-7, PPYEF-8 and PPYEF-9 belonging to seven genera in five families, three orders were isolated from the rhizomes. Except PPYEF-4 (Cladosporium sp.), other fungi could promote the embryo development of the P. polyphylla var. yunnanensis seeds, mostly reaching the extremely significant or significant level. PPYEF-9 (Trichoderma sp.) resulted in the highest embryo length and embryo-emerging ratio. [Conclusion] This paper could provide a reference for the application of the endophytic fungi of P. polyphylla var. yunnanensis in the dormancy-breaking of P. polyphylla var. yunnanensis seeds.展开更多
This study was conducted for the purposes of reducing consumption of wild resources of Paris polyphylla Smith, satisfying the market demand, saving the production cost, improving utilization rate of forest land, incre...This study was conducted for the purposes of reducing consumption of wild resources of Paris polyphylla Smith, satisfying the market demand, saving the production cost, improving utilization rate of forest land, increasing income of foresters and protecting ecological environment. Three types of fores{ land, i.e., the evergreen forest land （China fir forest）, deciduous forest land （walnut forest） and shrub land （honey suckle forest.） were selected, and seeds and cut rhizomes of Paris polyphylla Smith produced in Bamian Mountain, Hunan, were used in the study on understory seedling raising and cultivation techniques. The results showed that： when using different propagation materials, seed propagation could be adopted to reduce the consumption of wild resources and provide a large number of seedlings, and was beneficial to large-scale planting; and when conducing seedling and plantlet raising followed by transplanting and direct seeding and planting using different propagation materials in large-scale understory cultivation, direct seeding/ planting was appropriate for saving save production cost with the advantage of early production. Among the 3 different types of forest land, the best choice was broad- leaved forest land, which could achieve better economic benefit because of con- forming with the ecological habit and growth and development law of Paris poly- phylla Smith.展开更多
A novel phenylpropanoid glycosides 1, named parispolyside E and a novel derivation of phenolic glycoside 2, named parispolyside G, as well as two known flavonoid glycosides were isolated from the rhizome of Paris poly...A novel phenylpropanoid glycosides 1, named parispolyside E and a novel derivation of phenolic glycoside 2, named parispolyside G, as well as two known flavonoid glycosides were isolated from the rhizome of Paris polyphylla var. yunnanensis. Their structures were elucidaed by spectroscopic methods.展开更多
Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plant...Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plants age. We speculated that the concentrations of those components in the rhizomes are mediated by fungal endophytes. To test this hypothesis, we took both culture-dependent and-independent(metagenomics) approaches to analyze the communities of endophytic fungi that inhabit those rhizomes in plants of different age classes(four, six, and eight years old). In all, 147 isolates representing 18 fungal taxa were obtained from 270 segments(90 per age class). Based on morphological and genetic characteristics, Fusarium oxysporum(46.55% frequency of occurrence) was the predominant endophyte,followed by Leptodontidium sp.(8.66%) and Trichoderma viride(6.81%). Colonization of endophytic fungi was maximized in the eight-year-old rhizomes(33.33%) when compared with four-year-old(21.21%) and six-year-old(15.15%) rhizomes. Certain fungal species were present only at particular ages. For example,Alternaria sp., Cylindrocarpon sp., Chaetomium sp., Paraphaeosphaeria sporulosa, Pyrenochaeta sp., Penicillium swiecickii, T. viride, and Truncatella angustata were found only in the oldest plants. Analysis of(metagenomics) community DNA extracted from different-aged samples revealed that, at the class level,the majority of fungi had the highest sequence similarity to members of Sordariomycetes, followed by Eurotiomycetes and Saccharomycetes. These results were mostly in accord with those we obtained using culture methods. Fungal diversity and richness also changed over time. Our investigation is the first to show that the diversity of fungi in rhizomes of P. polyphylla var. yunnanensis is altered as plants age, and our findings provide a foundation for future examinations of useful compounds.展开更多
Paris polyphylla Smith var.yunnanensis(Franch.) Hand.-Mazz.is a rhizomatous,herbaceous,perennial plant that has been used for more than a thousand years in traditional Chinese medicine.It is facing extinction due to o...Paris polyphylla Smith var.yunnanensis(Franch.) Hand.-Mazz.is a rhizomatous,herbaceous,perennial plant that has been used for more than a thousand years in traditional Chinese medicine.It is facing extinction due to overharvesting.Steroids are the major therapeutic components in Paris roots,the commercial value of which increases with age.To date,no genomic data on the species have been available.In this study,transcriptome analysis of an 8-year-old root and a 4-year-old root provided insight into the metabolic pathways that generate the steroids.Using Illumina sequencing technology,we generated a high-quality sequence and demonstrated de novo assembly and annotation of genes in the absence of prior genome information.Approximately 87,577 unique sequences,with an average length of 614 bases,were obtained from the root cells.Using bioinformatics methods,we annotated approximately 65.51% of the unique sequences by conducting a similarity search with known genes in the National Center for Biotechnology Information's non-redundant database.The unique transcripts were functionally classified using the Gene Ontology hierarchy and the Kyoto Encyclopedia of Genes and Genomes database.Of 3082 genes that were identified as significantly differentially expressed between roots of different ages,1518(49.25%) were upregulated and 1564(50.75%) were downregulated in the older root.Metabolic pathway analysis predicted that 25 unigenes were responsible for the biosynthesis of the saponins steroids.These data represent a valuable resource for future genomic studies on this endangered species and will be valuable for efforts to genetically engineer P.polyphylla and facilitate saponin-rich plant development.展开更多
[Objectives]The analgesic and hemostatic effects of Paris polyphylla,Scutellaria baicalensis,and their compatibility were studied.[Methods](i)In the experiment on the writhing of mice induced by acetic acid,60 Kunming...[Objectives]The analgesic and hemostatic effects of Paris polyphylla,Scutellaria baicalensis,and their compatibility were studied.[Methods](i)In the experiment on the writhing of mice induced by acetic acid,60 Kunming SPF mice were randomly selected,and were randomly divided into 6 groups according to the male-to-female ratio of 1∶1,including blank control group,P.polyphylla group,S.baicalensis group,and three P.polyphylla and S.baicalensis compatibility groups(with the ratio of 1∶2,1∶1 and 1∶2).There were 10 mice in each group.In the experimental groups,the mice were given 1 g/mL Chinese medicine extract according to a dose of 20 mL/kg.In the control group,the mice were given 0.9%normal saline in equal volumes.Gavage was performed one time every 24 h and lasted for 14 d.1 h after the gavage on the 14th day,they were given 0.5%glacial acetic acid solution via intraperitoneal injection.(ii)In the hot plate experiment,60 Kunming SPF mice were selected,and the ratio of male to female,grouping,administration and gavage were the same as those of the glacial acetic acid-induced writhing experiment.Gavage was lasted for 14 d.1 h after the gavage on the 14th day,the mice were placed on a hot plate apparatus at(55±0.5)℃,and the time of licking hindfoot was measured.(iii)The hemostatic effect was explored through three experimental methods of tail hemostasis,femoral artery hemostasis and liver hemostasis.60 Kunming SPF mice were taken in each of the three experiments,and the male-female ratio and grouping were the same as above.The powder was covered on the surface of the wound sites,and the bleeding was observed.The hemostatic time was recorded,and the hemostatic time was recorded as 3 min if it exceeded 3 min.[Results](i)In the acetic acid-induced writhing experiment,compared with the blank control group,the pain of mice in the experimental groups was inhibited.Among them,P.polyphylla group,S.baicalensis group,compatibility 1∶2 group,compatibility 1∶1 group,and compatibility 2∶1 group had significant effects(P<0.05),and the inhibition rate of writhing was 20.43%,28.32%,52.30%,32.79%,and 39.02%,respectively.The analgesic effect of the compatibility 1∶2 group was the most obvious.(ii)In the hot plate experiment,compared with the blank control group,the experimental groups had analgesic effect.Among them,P.polyphylla group,S.baicalensis group,compatibility 1∶2 group,compatibility 1∶1 group,and compatibility 2∶1 group had significant effects(P<0.05),and the increase rate of pain threshold was 29.17%,47.83%,61.54%,and 50.61%,and 53.83%,respectively.The analgesic effect of the compatibility 2∶1 group was the most significant.(iii)In the hemostatic experiment,there was a significant difference in hemostatic time between the compatibility 1∶2 group and the blank control group,P.polyphylla group,and S.baicalensis group(P<0.05).The hemostatic effect of the compatibility 1∶2 group was the best.[Conclusions]P.polyphylla,S.baicalensis,and its compatibility had good analgesic and hemostatic effects,and the best compatibility ratio was 1∶2.展开更多
Phytochemical investigation of the rhizomes of Paris polyphylla var.stenophylla led to the isolation of two new highly oxygenated spirostanol saponins,named paristenosides A(1)and B(2),together with seven known compou...Phytochemical investigation of the rhizomes of Paris polyphylla var.stenophylla led to the isolation of two new highly oxygenated spirostanol saponins,named paristenosides A(1)and B(2),together with seven known compounds.Their structures were established mainly on the base of NMR spectroscopic techniques and mass spectrometry,as well as chemical methods.In addition,the cytotoxicity of the two new saponins was tested.Graphical Abstract Two new highly oxygenated spirostanol saponins,paristenosides A(1)and B(2),were isolated from the rhizomes of Paris polyphylla var.stenophylla.Their structures were established mainly based on NMR spectroscopic techniques and mass spectrometry,as well as chemical methods.展开更多
Paris polyphylla var.yunnanensis is a traditional Chinese medicinal plant,in which polyphyllin as the main medicinal component is an important secondary metabolite with bioactivity.Arbuscular mycorrhizal fungi(AMF)hav...Paris polyphylla var.yunnanensis is a traditional Chinese medicinal plant,in which polyphyllin as the main medicinal component is an important secondary metabolite with bioactivity.Arbuscular mycorrhizal fungi(AMF)have multiple positive effects on plants,while it is not clear whether AMF increase the content of medicinal components in medicinal plants.In this study,a total of nine AMF treatments were laid to analyze the mycorrhizal effect on polyphyllin accumulation and PpHMGR and PpSE expression of P.polyphylla var.yunnanensis.AMF increased the content of polyphyllin in the cultivated variety with low relation to the increase of inoculation intensity.Polyphyllin I,II,and VII were identified and partly improved by AMF inoculation,dependent on AMF treatments and culture environments.Similarly,the PpHMGR and PpSE expression was induced by mycorrhization,dependent on AMF species,whilst the induction was more obvious in PpSE than in PpHMGR after mycorrhization.It concluded that the symbiotic relationship between P.polyphylla var.yunnanensis and AMF increased polyphyllin content level in the plant,which was associated with the up-regulation of PpSE transcripts.展开更多
[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in roo...[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in root,stem and leaf from healthy and diseased plants. Infrared characteristic absorption peaks of healthy and diseased plants were found by infrared fingerprint method for comparative analysis.[Results]Total saponins content in root sample J4 of healthy plant reached 4. 89% and was the highest,while it was 0. 13% in stem sample J11 of healthy plant and was the lowest. Total saponins content in root sample B1 of diseased plant was 1. 68% and was the highest,while it was 0. 1% in stem sample B7 of diseased plant and was the lowest. In healthy and diseased plants,total saponins content in root was significantly higher than that in stem and leaf. Content of total saponins in root from healthy plant was significantly higher than that from diseased plant,and there was little difference in stem and leaf of healthy and diseased plants. Root sample Jg of healthy plant had obvious C-O characteristic vibration of polysaccharides and glycosides and absorption peak of O-H bending vibration in steroidal saponin near 1 160. 02 cm-1,while diseased plant had no obvious absorption peak. Near 861. 97 and 768. 07 cm-1,root sample Jg of healthy plant had significant C-C stretching vibration in sugar ring and characteristic absorption peak of characteristic vibration of steroidal saponin,while root sample Bg of diseased plant had no obvious characteristic peak. [Conclusions] Diseases significantly affected composition and content of total saponins and steroidal saponins from P. polyphylla var. chinensis,further affecting quality of P. polyphylla var. chinensis.展开更多
[Objectives] To determine the content of 10 kinds of inorganic elements(Pb,Cu,As,Cd,Hg,Zn,Fe,Mn,Sr and Cr) in samples of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( and polygerm varieties) produced ...[Objectives] To determine the content of 10 kinds of inorganic elements(Pb,Cu,As,Cd,Hg,Zn,Fe,Mn,Sr and Cr) in samples of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( and polygerm varieties) produced in different areas. [Methods] The wet digestion and technique of inductively coupled plasma optical emission spectrometry( ICP-OES) were used. [Results] Under the experimental conditions,elements were not related to each other,and several kinds of elements could be measured at the same time. The content of Pb exceeding the standard only in sample S4 of P. polyphylla Smith var. yunnanensis( and polygerm varieties),Hg was not detected in all samples,and the rest heavy metals did not exceed the limit. [Conclusions] This study was expected to provide a certain reference for the development and utilization of P. polyphylla Smith var. yunnanensis( and polygerm varieties).展开更多
A field experiment was conducted to determine the effects of two commercial strains composed of mulple arbuscular mycorrhizal fungi(AMF)species on plant growth,antioxidant capacity,and medicine quality of Paris polyph...A field experiment was conducted to determine the effects of two commercial strains composed of mulple arbuscular mycorrhizal fungi(AMF)species on plant growth,antioxidant capacity,and medicine quality of Paris polyphylla var.yunnanensis in three subtropical soils from Wanzhou,Anshun and Baoshan in fields.The results showed that AMF inoculation enhanced the fungal colonization rate and activities of both succinate dehydrogenase and alkaline phosphatase,thereby,enhancing the mycorrhizal viability of P.polyphylla var.yunnanensis.The concentrations of photosynthetic pigments(chlorophyll a,b,a+b and carotenoids),soluble sugar,soluble protein and photosynthetic capacity were higher in AMF-inoculated plants than in non-AMF-treated plants in field.AMFtreated plants recorded higher activities of catalase,peroxidase and superoxide dismutase,and caused the reduction in malondialdehyde content,indicating lower oxidative damage,compared with non-AMF plants.Polyphyllin I,Polyphyllin II,Polyphyllin III,Polyphyllin IV and total polyphyllin contents were increased by AMF treatment.In conclusion,AMF improved the plant growth,antioxidant capacity and medicinal quality of P.polyphylla var.yunnanensis seedlings.Hereinto,AMF effects on the soil from Wanzhou was relatively greater than on other soils.展开更多
[Objectives] The research aimed to identify Paris polyphylla with different growth year numbers and evaluate its resource quality.[Methods]Comparative research on P. polyphylla with different growth year numbers was c...[Objectives] The research aimed to identify Paris polyphylla with different growth year numbers and evaluate its resource quality.[Methods]Comparative research on P. polyphylla with different growth year numbers was conducted by using morphological identification and microscopic identification method. [Results] By contrasting original plant morphology,transverse sections of rhizome and fibrous root,powder,it displayed that microstructure of P. polyphylla had regularity difference in cell shape and size,duct quantity,trait and size,size and distribution of calcium oxalate cluster crystal. [Conclusions] The microstructure change of P. polyphylla was related to the number of its growth year,which could provide the reference for identifying the growth year number of P. polyphylla and evaluating its resource quality.展开更多
[Objectives] To make original plant and microscopic identification and study the traits of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( polygerm varieties) produced in Gaoligong Mountain areas,and pr...[Objectives] To make original plant and microscopic identification and study the traits of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( polygerm varieties) produced in Gaoligong Mountain areas,and provide experimental data for their identification and application. [Methods] The original plant powder was used,and paraffin sections and physicochemical properties were identified. [Results]P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas showed obvious microscopic features and physicochemical properties. [Conclusions] This study was expected to provide certain reference for identification of original plant and medicine and formulation of quality standard for P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas,and provided basis for their further research and development.展开更多
Paris polyphylla var. yunnanensis (Franch.), one of the best-known medicinal plants in China, has a dehiscent anther which physiologically work in pollination, however, the dehiscent anther always closes in response t...Paris polyphylla var. yunnanensis (Franch.), one of the best-known medicinal plants in China, has a dehiscent anther which physiologically work in pollination, however, the dehiscent anther always closes in response to darkness every day, and watering or raining every time. To explore this frequently closing and its unkown physiology, next-generation sequencing was performed, and the transcriptome was de novo assembled. RNA-sequencing was carried out in 15 samples including seven openning samples, four closed samples owing to darkness or watering, and tissue samples (leaf, petal, calyx, and stigma) were used for control. We obtained 72.75 GB data, assembled into 79,815 unigenes. Differentially expressed unigenes (DEGs) between opened and closed anther samples were 6231 and the DEGs between anther and control samples were 2831. Comparation between the two DEGs by KEGG enrichment showed that “plant hormone signal transduction” pathway is the most significant pathway for DEGs from closing anther vs. opening anther, and expression model of DEGs in the pathway might elicit change in germination and seed dormancy. Further examination of the action of the signal pathway on physiology showed “chromatin binding” function was prominent in “DNA binding” function of annotated DEGs between opened and closed anthers, of the 215 “chromatin binding” unigenes, 120 were involved in epigenetic silencing, and 50 of the epigenetic unigenes were directly related to germination or seed dormancy, strongly correlating anther closing to epigenetic modification and seed dormancy. These results were verified that at least three auxins involved in seed dormancy showed same expression patterns occurred in abnormal closing anther and seed embryo in Paris polyphylla var. yunnanensis. In conclusion, the information from transcriptome point out that frequent abnormal closing of dehiscent anthers possibly transfer the impact on seed dormancy, and epigenetic modification happened in closing may be the cause.展开更多
In this study,the resource distribution,morphological characteristics,artificial cultivation and market circulation of Paris Polyphylla var.yunnanensis and its polygerm varieties in Yunnan were investigated by referri...In this study,the resource distribution,morphological characteristics,artificial cultivation and market circulation of Paris Polyphylla var.yunnanensis and its polygerm varieties in Yunnan were investigated by referring to the research literature,field investigation and our planting experience.It is found that P.polyphylla var.yunnanensis is widely distributed in Yunnan Province,realizing large-scale artificial cultivation,while the wild resources of polygerm varieties are almost on the verge of extinction and are in urgent need of protection.展开更多
In this paper,the seedling raising,cultivation,field management,pest and disease control,harvesting and processing of Paris polyphylla Smith under the forest in Hunan Province were scientifically standardized,to provi...In this paper,the seedling raising,cultivation,field management,pest and disease control,harvesting and processing of Paris polyphylla Smith under the forest in Hunan Province were scientifically standardized,to provide technical support for standardized cultivation and sustainable development of Paris polyphylla Smith industry in Hunan Province.展开更多
Sugar-sugar glycosyltransferases play important roles in constructing complex and bioactive saponins.Here,we characterized a series of UDP-glycosyltransferases responsible for biosynthesizing the branched sugar chain ...Sugar-sugar glycosyltransferases play important roles in constructing complex and bioactive saponins.Here,we characterized a series of UDP-glycosyltransferases responsible for biosynthesizing the branched sugar chain of bioactive steroidal saponins from a widely known medicinal plant Paris polyphylla var.yunnanensis.Among them,a 2'-O-rhamnosyltransferase and three 6'-O-glucosyltrasferases catalyzed a cascade of glycosylation to produce steroidal diglycosides and triglycosides,respectively.These UDP-glycosyltransferases showed astonishing substrate promiscuity,resulting in the generation of a panel of 24 terpenoid glycosides including 15 previously undescribed compounds.A mutant library containing 44 variants was constructed based on the identification of critical residues by molecular docking simulations and protein model alignments,and a mutant UGT91AH1^(Y187A)with increased catalytic efficiency was obtained.The steroidal saponins exhibited remarkable antifungal activity against four widespread strains of human pathogenic fungi attributed to ergosterol-dependent damage of fungal cell membranes,and 2'-O-rhamnosylation appeared to correlate with strong antifungal effects.The findings elucidated the biosynthetic machinery for their production of steroidal saponins and revealed their potential as new antifungal agents.展开更多
基金supported by the CAMS Innovation Fund for Medical Sciences (CIFMS) (2017-I2M-3-013)the National Key Research and Development Program of China (Grant No. 2017YFC1700706)the National Natural Science Foundation of China (Grant No. 31471575).
文摘The mature seed of Paris polyphylla var.chinensis(PPC)is morphophysiologically dormant and develops differently under warm and cold temperatures.To elucidate the molecular mechanisms underlying temperature-dependent regulation of PPC seed dormancy and germination,we investigated the dynamic changes in PPC seed transcript levels under warm and cold temperature stratifications(WS and CS,respectively)by time-resolved RNA sequencing,focusing on genes related to hormone metabolism and signaling and cell wall remodeling(CWRM)and encoding transcription factors/regulators(TFs/TRs).A total of 48765 and 47836 differentially expressed genes(DEGs)were associated with WS and CS,respectively.Of these,17581 and 16652 DEGs,respectively,unique to WS and CS,and 5386 were common to both temperature stratifications across nine analyzed growth stages.The expression of hormone metabolism and signaling,TF/TR,and CWRM genes were closely associated with temperature.More genes related to gibberellin(GA),cytokinin,auxin,and brassinosteroid biosynthetic were upregulated in WS as compared to CS seeds,while genes related to dormancy release and germination were downregulated in WS seeds.However,only GA and abscisic acid levels were altered in PPC seeds breaking morphophysiological dormancy(MPD).Overall,37 TF and five TR families were upregulated whereas 24 TF and 16 TR families were downregulated in WS as compared to CS seeds.Most CWRM families were highly expressed under WS as compared to CS,suggesting that they promote endosperm weakening and embryo growth of WS seeds and facilitate MPD release and germination.A coexpression analysis revealed positive correlations between TFs/TRs and DEGs involved in hormone metabolism and signaling and CWRM.These results provided a basis for investigating the interaction between the endosperm and underdeveloped embryo in the regulation of PPC seed germination and seedling emergence.
基金supported by the Key R&D Program of Yunnan Province,China(Grant No.202103AC100003202101AS070228)+2 种基金the Major Special Project of the Ministry of Science and Technology(2021YFD10002022021YFD1601003)and the National Natural Science Foundation of China(Grant No.31860075),thank you.
文摘Arbuscular mycorrhizal fungi(AMF)are important members of the plant microbiome and affect the uptake and transfer of mineral elements by forming a symbiotic relationship with plant roots.Nitrogen(N),as an important mineral element,can directly affect plant growth and development at different N levels.It has been confirmed that inoculation with AMF can improve the efficiency of N utilization by plants.However,there are still fewer reports on the dynamic relationship between arbuscular mycorrhizal and plant secondary metabolites at different nitrogen levels.In this experiment,the physiological indexes and genes related to saponin synthesis were determined by applying different concentration gradients of nitrogen to the medicinal plant P.polyphylla var.yunnanensis infested by AMF as the test material.It was found that nitrogen addition increased the biomass,chlorophyll content,and nutrient content of above-and below-ground plant parts and increased the content of saponin content of P.polyphylla var.yunnanensis to some extent,but AMF inoculation increased the saponin content of P.polyphylla var.yunnanensis more significantly.AMF inoculation also promoted the expression of genes related to the saponin synthesis pathway,including 3-hydroxy-3-methylglutaryl coenzyme A synthase(HMGS),squalene epoxidase 1(SE1),and cycloartenol synthase(CAS),which is in according with the accumulation of saponin in plants.It also may increase the saponin content of AMF plants by altering the expression of P450s and UGTs related to saponin synthesis.
文摘[Objective] This study aimed to isolate the endophytic fungi of Paris polyphylla var. yunnanensis and investigate their effects on the embryo development of P. polyphylla var. yunnanensis seeds. [Method] The endophytic fungi of P. polyphylla were isolated and identified morphologically, and their effects on the embryo development of P. polyphylla var. yunnanensis seeds were studied by using paraffin sectioning and microphotography. [Result] Nine endophytic fungi, i.e. P. polyphylla var. yunnanensis endophytic fungi PPYEF-1, PPYEF-2, PPYEF-3, PPYEF-4, PPYEF-5, PPYEF-6, PPYEF-7, PPYEF-8 and PPYEF-9 belonging to seven genera in five families, three orders were isolated from the rhizomes. Except PPYEF-4 (Cladosporium sp.), other fungi could promote the embryo development of the P. polyphylla var. yunnanensis seeds, mostly reaching the extremely significant or significant level. PPYEF-9 (Trichoderma sp.) resulted in the highest embryo length and embryo-emerging ratio. [Conclusion] This paper could provide a reference for the application of the endophytic fungi of P. polyphylla var. yunnanensis in the dormancy-breaking of P. polyphylla var. yunnanensis seeds.
基金Supported by Central Financial Forestry Science and Technology Promotion project([2014]XT003)~~
文摘This study was conducted for the purposes of reducing consumption of wild resources of Paris polyphylla Smith, satisfying the market demand, saving the production cost, improving utilization rate of forest land, increasing income of foresters and protecting ecological environment. Three types of fores{ land, i.e., the evergreen forest land （China fir forest）, deciduous forest land （walnut forest） and shrub land （honey suckle forest.） were selected, and seeds and cut rhizomes of Paris polyphylla Smith produced in Bamian Mountain, Hunan, were used in the study on understory seedling raising and cultivation techniques. The results showed that： when using different propagation materials, seed propagation could be adopted to reduce the consumption of wild resources and provide a large number of seedlings, and was beneficial to large-scale planting; and when conducing seedling and plantlet raising followed by transplanting and direct seeding and planting using different propagation materials in large-scale understory cultivation, direct seeding/ planting was appropriate for saving save production cost with the advantage of early production. Among the 3 different types of forest land, the best choice was broad- leaved forest land, which could achieve better economic benefit because of con- forming with the ecological habit and growth and development law of Paris poly- phylla Smith.
文摘A novel phenylpropanoid glycosides 1, named parispolyside E and a novel derivation of phenolic glycoside 2, named parispolyside G, as well as two known flavonoid glycosides were isolated from the rhizome of Paris polyphylla var. yunnanensis. Their structures were elucidaed by spectroscopic methods.
基金supported by grants from the National Natural Science Foundation of China (81473310, 31260075, 31560085)
文摘Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plants age. We speculated that the concentrations of those components in the rhizomes are mediated by fungal endophytes. To test this hypothesis, we took both culture-dependent and-independent(metagenomics) approaches to analyze the communities of endophytic fungi that inhabit those rhizomes in plants of different age classes(four, six, and eight years old). In all, 147 isolates representing 18 fungal taxa were obtained from 270 segments(90 per age class). Based on morphological and genetic characteristics, Fusarium oxysporum(46.55% frequency of occurrence) was the predominant endophyte,followed by Leptodontidium sp.(8.66%) and Trichoderma viride(6.81%). Colonization of endophytic fungi was maximized in the eight-year-old rhizomes(33.33%) when compared with four-year-old(21.21%) and six-year-old(15.15%) rhizomes. Certain fungal species were present only at particular ages. For example,Alternaria sp., Cylindrocarpon sp., Chaetomium sp., Paraphaeosphaeria sporulosa, Pyrenochaeta sp., Penicillium swiecickii, T. viride, and Truncatella angustata were found only in the oldest plants. Analysis of(metagenomics) community DNA extracted from different-aged samples revealed that, at the class level,the majority of fungi had the highest sequence similarity to members of Sordariomycetes, followed by Eurotiomycetes and Saccharomycetes. These results were mostly in accord with those we obtained using culture methods. Fungal diversity and richness also changed over time. Our investigation is the first to show that the diversity of fungi in rhizomes of P. polyphylla var. yunnanensis is altered as plants age, and our findings provide a foundation for future examinations of useful compounds.
基金supported by the National Natural Science Foundation of China(81473310,31260075,31560085)
文摘Paris polyphylla Smith var.yunnanensis(Franch.) Hand.-Mazz.is a rhizomatous,herbaceous,perennial plant that has been used for more than a thousand years in traditional Chinese medicine.It is facing extinction due to overharvesting.Steroids are the major therapeutic components in Paris roots,the commercial value of which increases with age.To date,no genomic data on the species have been available.In this study,transcriptome analysis of an 8-year-old root and a 4-year-old root provided insight into the metabolic pathways that generate the steroids.Using Illumina sequencing technology,we generated a high-quality sequence and demonstrated de novo assembly and annotation of genes in the absence of prior genome information.Approximately 87,577 unique sequences,with an average length of 614 bases,were obtained from the root cells.Using bioinformatics methods,we annotated approximately 65.51% of the unique sequences by conducting a similarity search with known genes in the National Center for Biotechnology Information's non-redundant database.The unique transcripts were functionally classified using the Gene Ontology hierarchy and the Kyoto Encyclopedia of Genes and Genomes database.Of 3082 genes that were identified as significantly differentially expressed between roots of different ages,1518(49.25%) were upregulated and 1564(50.75%) were downregulated in the older root.Metabolic pathway analysis predicted that 25 unigenes were responsible for the biosynthesis of the saponins steroids.These data represent a valuable resource for future genomic studies on this endangered species and will be valuable for efforts to genetically engineer P.polyphylla and facilitate saponin-rich plant development.
基金Major Research and Development Plan Program of Guangxi,China(Guike AB18221095)Famous Teacher Training Plan Program of Youjiang Medical University for Nationalities——National-level Famous Teacher Training Plan Program(Youyiyuanzi[2018]No.98)+1 种基金High-level Talent Research Project of Youjiang Medical University for Nationalities(01002018079)National and Autonomous Region Innovation Planning Project for University Students in 2019(201910599023).
文摘[Objectives]The analgesic and hemostatic effects of Paris polyphylla,Scutellaria baicalensis,and their compatibility were studied.[Methods](i)In the experiment on the writhing of mice induced by acetic acid,60 Kunming SPF mice were randomly selected,and were randomly divided into 6 groups according to the male-to-female ratio of 1∶1,including blank control group,P.polyphylla group,S.baicalensis group,and three P.polyphylla and S.baicalensis compatibility groups(with the ratio of 1∶2,1∶1 and 1∶2).There were 10 mice in each group.In the experimental groups,the mice were given 1 g/mL Chinese medicine extract according to a dose of 20 mL/kg.In the control group,the mice were given 0.9%normal saline in equal volumes.Gavage was performed one time every 24 h and lasted for 14 d.1 h after the gavage on the 14th day,they were given 0.5%glacial acetic acid solution via intraperitoneal injection.(ii)In the hot plate experiment,60 Kunming SPF mice were selected,and the ratio of male to female,grouping,administration and gavage were the same as those of the glacial acetic acid-induced writhing experiment.Gavage was lasted for 14 d.1 h after the gavage on the 14th day,the mice were placed on a hot plate apparatus at(55±0.5)℃,and the time of licking hindfoot was measured.(iii)The hemostatic effect was explored through three experimental methods of tail hemostasis,femoral artery hemostasis and liver hemostasis.60 Kunming SPF mice were taken in each of the three experiments,and the male-female ratio and grouping were the same as above.The powder was covered on the surface of the wound sites,and the bleeding was observed.The hemostatic time was recorded,and the hemostatic time was recorded as 3 min if it exceeded 3 min.[Results](i)In the acetic acid-induced writhing experiment,compared with the blank control group,the pain of mice in the experimental groups was inhibited.Among them,P.polyphylla group,S.baicalensis group,compatibility 1∶2 group,compatibility 1∶1 group,and compatibility 2∶1 group had significant effects(P<0.05),and the inhibition rate of writhing was 20.43%,28.32%,52.30%,32.79%,and 39.02%,respectively.The analgesic effect of the compatibility 1∶2 group was the most obvious.(ii)In the hot plate experiment,compared with the blank control group,the experimental groups had analgesic effect.Among them,P.polyphylla group,S.baicalensis group,compatibility 1∶2 group,compatibility 1∶1 group,and compatibility 2∶1 group had significant effects(P<0.05),and the increase rate of pain threshold was 29.17%,47.83%,61.54%,and 50.61%,and 53.83%,respectively.The analgesic effect of the compatibility 2∶1 group was the most significant.(iii)In the hemostatic experiment,there was a significant difference in hemostatic time between the compatibility 1∶2 group and the blank control group,P.polyphylla group,and S.baicalensis group(P<0.05).The hemostatic effect of the compatibility 1∶2 group was the best.[Conclusions]P.polyphylla,S.baicalensis,and its compatibility had good analgesic and hemostatic effects,and the best compatibility ratio was 1∶2.
基金This work is funded by the National Natural Science Funding of China(Grand No.31570363)the Key Research Program of the Chinese Academy of Sciences(Grand No.KSZDEW-Z-004-03-4)the Natural Science Foundation of Yunnan Province(Grand No.2015FA031).
文摘Phytochemical investigation of the rhizomes of Paris polyphylla var.stenophylla led to the isolation of two new highly oxygenated spirostanol saponins,named paristenosides A(1)and B(2),together with seven known compounds.Their structures were established mainly on the base of NMR spectroscopic techniques and mass spectrometry,as well as chemical methods.In addition,the cytotoxicity of the two new saponins was tested.Graphical Abstract Two new highly oxygenated spirostanol saponins,paristenosides A(1)and B(2),were isolated from the rhizomes of Paris polyphylla var.stenophylla.Their structures were established mainly based on NMR spectroscopic techniques and mass spectrometry,as well as chemical methods.
基金supported by the National Natural Science Foundation of China(No.81260622)Chongqing Natural Science Foundation Project(cstc2018jcyjAX0267).
文摘Paris polyphylla var.yunnanensis is a traditional Chinese medicinal plant,in which polyphyllin as the main medicinal component is an important secondary metabolite with bioactivity.Arbuscular mycorrhizal fungi(AMF)have multiple positive effects on plants,while it is not clear whether AMF increase the content of medicinal components in medicinal plants.In this study,a total of nine AMF treatments were laid to analyze the mycorrhizal effect on polyphyllin accumulation and PpHMGR and PpSE expression of P.polyphylla var.yunnanensis.AMF increased the content of polyphyllin in the cultivated variety with low relation to the increase of inoculation intensity.Polyphyllin I,II,and VII were identified and partly improved by AMF inoculation,dependent on AMF treatments and culture environments.Similarly,the PpHMGR and PpSE expression was induced by mycorrhization,dependent on AMF species,whilst the induction was more obvious in PpSE than in PpHMGR after mycorrhization.It concluded that the symbiotic relationship between P.polyphylla var.yunnanensis and AMF increased polyphyllin content level in the plant,which was associated with the up-regulation of PpSE transcripts.
基金Supported by the National Key R&D Program(2018YFC1708005)the Sichuan Science and Technology Plan Project(2018JY0111)the Innovating Research Program of Postgraduates of Southwest Minzu University in 2018(cx2018sz83)
文摘[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in root,stem and leaf from healthy and diseased plants. Infrared characteristic absorption peaks of healthy and diseased plants were found by infrared fingerprint method for comparative analysis.[Results]Total saponins content in root sample J4 of healthy plant reached 4. 89% and was the highest,while it was 0. 13% in stem sample J11 of healthy plant and was the lowest. Total saponins content in root sample B1 of diseased plant was 1. 68% and was the highest,while it was 0. 1% in stem sample B7 of diseased plant and was the lowest. In healthy and diseased plants,total saponins content in root was significantly higher than that in stem and leaf. Content of total saponins in root from healthy plant was significantly higher than that from diseased plant,and there was little difference in stem and leaf of healthy and diseased plants. Root sample Jg of healthy plant had obvious C-O characteristic vibration of polysaccharides and glycosides and absorption peak of O-H bending vibration in steroidal saponin near 1 160. 02 cm-1,while diseased plant had no obvious absorption peak. Near 861. 97 and 768. 07 cm-1,root sample Jg of healthy plant had significant C-C stretching vibration in sugar ring and characteristic absorption peak of characteristic vibration of steroidal saponin,while root sample Bg of diseased plant had no obvious characteristic peak. [Conclusions] Diseases significantly affected composition and content of total saponins and steroidal saponins from P. polyphylla var. chinensis,further affecting quality of P. polyphylla var. chinensis.
基金Supported by Key Sci-Tech Project of Sichuan Provincial Science and Technology Department(2016FZ0045)Fundamental Research Funds for the Central Universities of Southwest Minzu University(2018NZD10)
文摘[Objectives] To determine the content of 10 kinds of inorganic elements(Pb,Cu,As,Cd,Hg,Zn,Fe,Mn,Sr and Cr) in samples of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( and polygerm varieties) produced in different areas. [Methods] The wet digestion and technique of inductively coupled plasma optical emission spectrometry( ICP-OES) were used. [Results] Under the experimental conditions,elements were not related to each other,and several kinds of elements could be measured at the same time. The content of Pb exceeding the standard only in sample S4 of P. polyphylla Smith var. yunnanensis( and polygerm varieties),Hg was not detected in all samples,and the rest heavy metals did not exceed the limit. [Conclusions] This study was expected to provide a certain reference for the development and utilization of P. polyphylla Smith var. yunnanensis( and polygerm varieties).
基金This work was supported by the National Natural Science Foundation of China(81260622)Applied Basic Research Program of Yunnan Province(2011FB081)Scientific Research Fund Key Project of Yunnan Province(2012Z119).
文摘A field experiment was conducted to determine the effects of two commercial strains composed of mulple arbuscular mycorrhizal fungi(AMF)species on plant growth,antioxidant capacity,and medicine quality of Paris polyphylla var.yunnanensis in three subtropical soils from Wanzhou,Anshun and Baoshan in fields.The results showed that AMF inoculation enhanced the fungal colonization rate and activities of both succinate dehydrogenase and alkaline phosphatase,thereby,enhancing the mycorrhizal viability of P.polyphylla var.yunnanensis.The concentrations of photosynthetic pigments(chlorophyll a,b,a+b and carotenoids),soluble sugar,soluble protein and photosynthetic capacity were higher in AMF-inoculated plants than in non-AMF-treated plants in field.AMFtreated plants recorded higher activities of catalase,peroxidase and superoxide dismutase,and caused the reduction in malondialdehyde content,indicating lower oxidative damage,compared with non-AMF plants.Polyphyllin I,Polyphyllin II,Polyphyllin III,Polyphyllin IV and total polyphyllin contents were increased by AMF treatment.In conclusion,AMF improved the plant growth,antioxidant capacity and medicinal quality of P.polyphylla var.yunnanensis seedlings.Hereinto,AMF effects on the soil from Wanzhou was relatively greater than on other soils.
基金Supported by Innovative Research Project of Graduate Student of Southwest Minzu University(CX2017SZ089)
文摘[Objectives] The research aimed to identify Paris polyphylla with different growth year numbers and evaluate its resource quality.[Methods]Comparative research on P. polyphylla with different growth year numbers was conducted by using morphological identification and microscopic identification method. [Results] By contrasting original plant morphology,transverse sections of rhizome and fibrous root,powder,it displayed that microstructure of P. polyphylla had regularity difference in cell shape and size,duct quantity,trait and size,size and distribution of calcium oxalate cluster crystal. [Conclusions] The microstructure change of P. polyphylla was related to the number of its growth year,which could provide the reference for identifying the growth year number of P. polyphylla and evaluating its resource quality.
基金Supported by Key Sci-Tech Project of Sichuan Provincial Science and Technology Department(2016FZ0045)Fundamental Research Funds for the Central Universities of Southwest Minzu University(2018NZD10)
文摘[Objectives] To make original plant and microscopic identification and study the traits of Paris polyphylla Smith var. yunnanensis( Franch.) Hand.-Mazz.( polygerm varieties) produced in Gaoligong Mountain areas,and provide experimental data for their identification and application. [Methods] The original plant powder was used,and paraffin sections and physicochemical properties were identified. [Results]P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas showed obvious microscopic features and physicochemical properties. [Conclusions] This study was expected to provide certain reference for identification of original plant and medicine and formulation of quality standard for P. polyphylla Smith var. yunnanensis( polygerm varieties) produced in Gaoligong Mountain areas,and provided basis for their further research and development.
文摘Paris polyphylla var. yunnanensis (Franch.), one of the best-known medicinal plants in China, has a dehiscent anther which physiologically work in pollination, however, the dehiscent anther always closes in response to darkness every day, and watering or raining every time. To explore this frequently closing and its unkown physiology, next-generation sequencing was performed, and the transcriptome was de novo assembled. RNA-sequencing was carried out in 15 samples including seven openning samples, four closed samples owing to darkness or watering, and tissue samples (leaf, petal, calyx, and stigma) were used for control. We obtained 72.75 GB data, assembled into 79,815 unigenes. Differentially expressed unigenes (DEGs) between opened and closed anther samples were 6231 and the DEGs between anther and control samples were 2831. Comparation between the two DEGs by KEGG enrichment showed that “plant hormone signal transduction” pathway is the most significant pathway for DEGs from closing anther vs. opening anther, and expression model of DEGs in the pathway might elicit change in germination and seed dormancy. Further examination of the action of the signal pathway on physiology showed “chromatin binding” function was prominent in “DNA binding” function of annotated DEGs between opened and closed anthers, of the 215 “chromatin binding” unigenes, 120 were involved in epigenetic silencing, and 50 of the epigenetic unigenes were directly related to germination or seed dormancy, strongly correlating anther closing to epigenetic modification and seed dormancy. These results were verified that at least three auxins involved in seed dormancy showed same expression patterns occurred in abnormal closing anther and seed embryo in Paris polyphylla var. yunnanensis. In conclusion, the information from transcriptome point out that frequent abnormal closing of dehiscent anthers possibly transfer the impact on seed dormancy, and epigenetic modification happened in closing may be the cause.
基金Sichuan Provincial Department of Science and Technology(2017SZYZF0004).
文摘In this study,the resource distribution,morphological characteristics,artificial cultivation and market circulation of Paris Polyphylla var.yunnanensis and its polygerm varieties in Yunnan were investigated by referring to the research literature,field investigation and our planting experience.It is found that P.polyphylla var.yunnanensis is widely distributed in Yunnan Province,realizing large-scale artificial cultivation,while the wild resources of polygerm varieties are almost on the verge of extinction and are in urgent need of protection.
基金Supported by Hunan Traditional Chinese Medicine Research Project (2021133)Technology System of Modern Agricultural Chinese Herbal Medicine Industry in Hunan Province
文摘In this paper,the seedling raising,cultivation,field management,pest and disease control,harvesting and processing of Paris polyphylla Smith under the forest in Hunan Province were scientifically standardized,to provide technical support for standardized cultivation and sustainable development of Paris polyphylla Smith industry in Hunan Province.
基金supported financially by the National Natural Science Foundation of China (Nos.82222072 and U1902214)Yunnan Key Research and Development Program (No.2019ZF011-2,China)the Research Project of Sichuan Province (2022JDJQ0055,China)。
文摘Sugar-sugar glycosyltransferases play important roles in constructing complex and bioactive saponins.Here,we characterized a series of UDP-glycosyltransferases responsible for biosynthesizing the branched sugar chain of bioactive steroidal saponins from a widely known medicinal plant Paris polyphylla var.yunnanensis.Among them,a 2'-O-rhamnosyltransferase and three 6'-O-glucosyltrasferases catalyzed a cascade of glycosylation to produce steroidal diglycosides and triglycosides,respectively.These UDP-glycosyltransferases showed astonishing substrate promiscuity,resulting in the generation of a panel of 24 terpenoid glycosides including 15 previously undescribed compounds.A mutant library containing 44 variants was constructed based on the identification of critical residues by molecular docking simulations and protein model alignments,and a mutant UGT91AH1^(Y187A)with increased catalytic efficiency was obtained.The steroidal saponins exhibited remarkable antifungal activity against four widespread strains of human pathogenic fungi attributed to ergosterol-dependent damage of fungal cell membranes,and 2'-O-rhamnosylation appeared to correlate with strong antifungal effects.The findings elucidated the biosynthetic machinery for their production of steroidal saponins and revealed their potential as new antifungal agents.
