Chemical constituents from Paris rugosa rhizomes and their antimicrobial activities / 中国中药杂志

Paris rugosa(Melanthiaceae) only grows in Yunnan province of China at present, and its chemical constituents have not been systematically studied. In this study, nine compounds, including one new compound pariposide G(1) and eight known compounds of cerin(2), stigmast-4-en-3-one(3), β-ecdysone(4), ophiopogonin C'(5), methyl protogracillin(6), gracillin(7), parissaponin H(8), and parisyunnanoside G(9), were isolated and identified from the ethanol extract of P. rugosa rhizomes by column chromatography methods and semi-preparative high-performance liquid chromatography(HPLC). Compounds 1-9 were isolated from this plant for the first time. The antibacterial and antifungal activities of all the compounds were evaluated. The results showed that ophiopogonin C' had strong inhibitory effects on Candida albicans [MIC_(90)=(4.68±0.01) μmol·L~(-1)] and the fluconazole-resistant strain of C. albicans [MIC_(90)=(4.66±0.02) μmol·L~(-1)].

Modeling of Brazilian Carbon Dioxide Emissions: a Review

Abstract: Brazil is a signatory to the Paris Agreement and aims to reduce 43% of CO2 emissions by 2030, compared to 2005. However, changes in energy policies are needed to achieve this goal, evaluating the produced effects on emissions. One way to predict these effects is through mathematical modeling. In this paper, we carried out a literature review to identify the most used model types and independent variables to forecasting Brazilian CO2 emissions. The review showed that gray models and artificial neural networks are the most used ones. Furthermore, we also identified that economic growth and energy consumption are the main independent variables.

Effect of Potassium Solubilizing Bacteria on Rhizosphere Soil Microenvironment of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

ObjectiveTo investigate the effects of the inoculation of potassium-solubilizing bacteria on the rhizosphere soil microenvironment of Paris polyphylla var. yunnanensis. MethodThe effects of different potassium-solubilizing bacteria on the physical and chemical properties， microbial community structure， and soil enzyme activity in the rhizosphere soil of P. polyphylla var. yunnanensis were investigated by pot planting at room temperature. The correlation of various indexes was analyzed. ResultThe inoculation with different potassium-solubilizing bacteria could significantly affect the physical and chemical properties of rhizosphere soil of P. polyphylla var. yunnanensis. The mass fractions of available nitrogen， available phosphorus， and available potassium were 24.5-90.5 mg·kg-1， 2.53-25.9 mg·kg-1， and 132-312 mg·kg-1， respectively， and the soil pH was 7.08-7.75， which were in line with the optimal ranges of P. polyphylla var. yunnanensis planting. The inoculation of different potassium-solubilizing bacteria could affect the number of bacteria， actinomycetes， and fungi in rhizosphere soil to varying degrees. The transformation of soil from "fungal type" to "bacterial type" marks the improvement of soil fertility. It also affected the enzyme activity of rhizosphere soil， and the activities of neutral phosphatase， protease， and polyphenol oxidase showed an increasing trend. The correlation analysis showed that the number of bacteria was negatively correlated with the number of fungi （r=-0.856， P<0.01）， positively correlated with the number of actinomycetes， the content of available nitrogen and available potassium， and negatively correlated with soil pH. ConclusionThe inoculation of potassium-solubilizing bacteria can effectively improve the content of available potassium， available nitrogen， available phosphorus， and other nutrients in the rhizosphere soil of P. polyphylla var. yunnanensis， improve soil fertility， alleviate the continuous cropping obstacles of P. polyphylla var. yunnanensis， and lay a theoretical foundation for the green and sustainable development of P. polyphylla var. yunnanensis.

Effect of Organophosphate-Solubilizing Bacteria on Photosynthesis， Physiology， and Biochemistry of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

ObjectiveTo study the effect of organophosphate-solubilizing bacteria and compound bacteria on the photosynthesis and physiological and biochemical characteristics of leaves of Paris polyphylla var. yunnanensis， and to provide a reference for selecting suitable bacterial fertilizers in artificial cultivation of this medicinal species. MethodPot experiment was carried out indoor and the following groups were designed： control （CK）， inoculation with Bacillus mycoides （S1）， inoculation with B. wiedmannii （S2）， inoculation with B. proteolyticus （S3）， inoculation with B. mycoides and B. wiedmannii （S4）， inoculation with B. mycoides and B. proteolyticus （S5）， inoculation with B. wiedmannii and B. proteolyticus （S6）， and inoculation with B. mycoides， B. wiedmannii and B. proteolyticus （S7）. Then， the growth and development， photosynthesis， and various physiological and biochemical indexes of the leaves of this species were observed. ResultCompared with CK， the treatment groups showed decrease in content of malondialdehyde in the leaves （P<0.05）， particularly S7 （content was only about 1/3 that of the CK）. The leaf area， photosynthetic parameters， photosynthetic pigment content， soluble sugar content， soluble protein content， and activity of superoxide dismutase （SOD）， peroxidase （POD）， and catalase （CAT） in leaves of the treatment groups were all improved. Among them， the leaf area， soluble sugar content， and soluble protein content were the highest in S7， which were 2.8， 2.1， and 2.2 times that of the CK， respectively. SOD activity peaked in S6 （2.9 times higher than that in the CK） and the highest activity of POD and CAT was detected in S5 （1.5 times and 2.1 times， respectively higher than that in the CK）. ConclusionInoculation with different organophosphate-solubilizing bacteria or compound bacteria can promote the growth and development of P. polyphylla var. yunnanensis and improve its resistance to stresses. The combination of B. mycoides and B. proteolyticus and the combination of the three achieved the have the best effect. This study provides a reference for the selection of bacterial fertilizers for artificial cultivation of P. polyphylla var. yunnanensis.

Effect of Inoculation with Organophosphate Solubilizing Bacteria on Quality and Soil Fertility of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

ObjectiveThe effect of inoculation with different organophosphate-resolving bacteria or compound bacteria on the quality of Paris polyphylla var. yunnanensis medicinal materials and rhizosphere soil fertility were studied to provide a reference for the development and application of biological bacterial fertilizer in artificial cultivation of P. polyphylla var. yunnanensis. MethodThe three dominant species of organophosphate-solubilizing bacteria were inoculated separately and in combination in sterilized soil by single-factor indoor pot planting， and no inoculation was used as the control group. The effect of inoculation of organophosphate-solubilizing bacteria on total saponins content in rhizomes of P. polyphylla var. yunnanensis， as well as microbial numbers， enzyme activities and nutrient contents in rhizosphere soil were analyzed. ResultIn the seven treatments inoculated with organophosphate-solubilizing bacteria， the total saponin content in the rhizomes of Paris polyphylla var. yunnanensis was increased by 16.42%， 3.83%， 16.86%， 33.69%， 2.11%， 13.44%， and 28.83%， respectively， compared with the control. Inoculation with organophosphate-solubilizing bacteria increased the number of soil microorganisms to varying degrees， and the effects of S6 and S7 treatments were the most significant. Inoculation with organophosphate-solubilizing bacteria improved the enzyme activity， and the effect of S7 treatment was the most significant. The activities of acid phosphatase， neutral phosphatase， alkaline phosphatase， protease， invertase and catalase were 49.96% and 104.67% ， 110.17%， 99.61%， 26.26%， 11.29% higher than those of the control， respectively. Inoculation with organophosphate-solubilizing bacteria reduced the pH of the rhizosphere soil and increased the content of soil available nutrients. Under the S7 treatment， the contents of alkaline hydrolyzable nitrogen， available phosphorus and available potassium in the rhizosphere soil of P. polyphylla var. yunnanensis were 181.46%， 51.64% and 42.62% higher than those of the control， respectively. Correlation analysis showed that there was a significant positive correlation between total saponins and phosphatase activities， a significant positive correlation between soil microorganisms and soil enzyme activities， and a very significant positive correlation between soil nutrients. ConclusionInoculation of different organophosphate-resolving bacteria or compound bacteria can improve the quality and rhizosphere soil fertility of P. polyphylla var. yunnanensis. Among them， the mixed inoculation of three kinds of bacteria and the mixed inoculation of B. mycoides and B. wiedmannii have the best effect.

Effect of Potassium Solubilizing Bacteria on Leaf Area，Photosynthetic Pigment Content，Physiology and Biochemistry of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

ObjectiveTo investigate the effects of inoculation of potassium-solubilizing bacteria on the physiological characteristics of Paris polyphylla var. yunnanensis. MethodThe effects of different potassium-solubilizing bacteria on leaf area，photosynthetic pigment content， and active component indexes of P. polyphylla var. yunnanensis were investigated by pot planting at room temperature. ResultThe results showed that the inoculation of potassium-solubilizing bacteria could increase the leaf area of P. polyphylla var. yunnanensis. The largest average leaf area was observed in the S3 group，reaching 10.497 cm2，with a maximum increase of 11.0% compared with that of the CK group. The inoculation of potassium-solubilizing bacteria could increase the content of photosynthetic pigments in leaves. Among them，the increase in chlorophyll a content was significant （P<0.05），which was 80.6% higher than that of the CK group. The chlorophyll a/b values in all treatment groups were higher than that of the CK group. The activities of superoxide dismutase（SOD），catalase（CAT） and peroxidase（POD） in the leaves of P. polyphylla var. yunnanensis increased by 88.4%，33.8%， and 30.6%，respectively， as compared with that in the CK group. The inoculation of potassium-solubilizing bacteria could promote the accumulation of osmoregulation substances such as soluble sugar and soluble protein in leaf cells of P. polyphylla var. yunnanensis，with a maximum increase of 55.3% and 70.5% respectively compared with that of the CK group. Meanwhile，it also reduced the content of malondialdehyde（MDA），with a maximum decrease of 34.1% compared with that of the CK group. The results of correlation analysis showed that there were some correlations between different chemical components in P. polyphylla var. yunnanensis leaves. ConclusionThe inoculation of potassium-solubilizing bacteria can effectively promote the growth and development of P. polyphylla var. yunnanensis and improve its survival ability in adversity. It is of practical scientific significance and potential application value for improving the survival rate of P. polyphylla var. yunnanensis by artificial planting.

Effects of Plant Growth-promoting Rhizobacteria on Physiological Characteristics and Inorganic Elements in Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

ObjectiveTo study the effects of different plant growth-promoting rhizobacteria （PGPR） on the growth of Paris polyphylla var. yunnanensis seedlings and the quality of its medicinal parts， in order to provide reference for the cultivation of high-quality P. polyphylla var. yunnanensis. MethodThe pot culture experiment at room temperature and the single-factor completely random design were employed for exploring the effects of five PGPR on physiological characteristics and inorganic elements of P. polyphylla var. yunnanensis. ResultThe results showed that the exogenous inoculation of different PGPR promoted the growth and development of P. polyphylla var. yunnanensis to varying degrees， delayed the senescence of leaves， and improved the medicinal value of new and old rhizomes. Compared with the non-inoculated control， the exogenous inoculation of compound microbial fertilizer （FH） and microbial agent Sanju Guanjin liquid （SJ） enhanced the root vigor， increased the content of photosynthetic pigments and the activities of anti-oxidant enzymes ［superoxide dismutase （SOD）， catalase （CAT）， and peroxidase （POD）］， and reduced the content of malondialdehyde （MDA） in leaves. Their inhibition rates against MDA were 10.46%-39.62% and 20.99%-53.12%， respectively. With the growth of P. polyphylla var. yunnanensis， the inhibition rate against MDA gradually increased， which effectively delayed the senescence of P. polyphylla var. yunnanensis leaves. In addition， the exogenous inoculation of different PGPR promoted the accumulation of nutrient elements in new and old rhizomes， lowered the heavy metal content to varying degrees， and improved the medicinal value of P. polyphylla var. yunnanensis rhizomes. ConclusionFH and SJ have exhibited the best promoting effect on the growth of P. polyphylla var. yunnanensis seedlings and also the best regulatory effect on the medicinal value of P. polyphylla var. yunnanensis rhizomes， which has provided reference for the application and promotion of PGPR in the growth of P. polyphylla var. yunnanensis.

Spatial distribution characteristics of metabolities in rhizome of Paris polyphylla var. yunnanensis: based on MALDI-MSI / 中国中药杂志

In this study, a method was established for in-situ visualization of metabolite distribution in the rhizome of Paris polyphylla var. yunnanensis. To be specific, through matrix-assisted laser desorption/ionization-mass spectrometry imaging(MALDI-MSI), the spatial locations of steroidal saponins, amino acids, organic acids, phytosterols, phytoecdysones, nucleosides, and esters in rhizome of the medicinal plant were directly analyzed, and six unknown compounds with differential distribution in rhizome tissues were identified. The specific procedure is as follows: preparation of rhizome tissue section, matrix screening and optimization, and MALDI-MSI analysis. The results showed that the steroidal saponins were mainly distributed in the central, amino acids in epidermis and cortex, low-molecular-weight organic acids in central epidermis, phytosterols in the epidermis and lateral cortex, the phytoecdysones in epidermis and cortex, nucleosides(uneven distribution) in epidermis and cortex, growth hormones around the epidermis and cortex, particularly outside the cortex, and esters in cortex with unobvious difference among different tissues. In this study, the spatial distribution of meta-bolites in the rhizome of P. polyphylla var. yunnanensis was characterized for the first time. The result can serve as a reference for identifying and extracting endogenous metabolites of P. polyphylla var. yunnanensis, exploring the synthesis and metabolism mechanisms of the metabolites, and evaluating the quality of medicinal materials.

Separation and structural elucidation of C25 epimers of furostanol saponin from aerial parts of Paris polyphylla var. chinensis / 中国中药杂志

Paris polyphylla var. chinensis(PPC) is used as one of the origin plants of Paridis Rhizoma described in the Chinese Pharmacopoeia(2020 edition). Its resources shortage makes the planting scale gradually expand, and plenty of aerial parts are abandoned because of not being effectively used. On the basis of previous research, this study separated steroidal saponins to further clarify the chemical composition of the aerial parts of PPC. As a result, three pairs of 25R or 25S epimers of furostanol saponins were obtained by various column chromatography techniques. Their structures were identified as neosolanigroside Y6(1), solanigroside Y6(2), neoprotogracillin(3), protogracillin(4), neoprotodioscin(5) and protodioscin(6) by spectral data combining with chemical transformation. Compound 1 is a new compound, and compounds 2, 3 and 5 are isolated from Paris plants for the first time. Compounds 4 and 6 are isolated from this plant for the first time. Previously, only several spirostanol glycosides with 25S configuration were isolated from Paris plants. Guided by mass spectrometry, the present study isolated the furostanol saponins with 25S configuration from this genus for the first time, which further enriches the chemical information of Paris genus and provides a reference for the isolation of similar compounds.

Qualitative and quantitative analysis of Paris polyphylla var. chinensis by UPLC-Q-TOF-MS/MS and HPLC / 中国中药杂志

Paridis Rhizoma(PR) is prepared from the dried rhizome of Paris polyphylla var. yunnanensis(PPY) or P. polyphylla var. chinensis(PPC) in Liliaceae family. The rapid development of PPY or PPC planting industry resulted from resource shortage has caused the waste of a large number of non-medicinal resources. To clarify the chemical compositions in rhizomes, fibrous roots, stems, leaves, seeds and pericarps of PPC, and explore the comprehensive application value and development prospect of these parts, the qualitative and quantitative analyses on the different parts of PPC were carried out by ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) and high performance liquid chromatography(HPLC). A total of 136 compounds were identified, including 112 steroidal saponins, 6 flavonoids, 11 nitrogen-containing compounds and 7 phytosterols. Rhizomes, fibrous roots, and seeds mainly contained protopennogenyl glycosides and pennogenyl glycosides; leaves and stems mainly contained protodiosgenyl glycosides and diosgenyl glycosides; pericarps mainly contained pennogenyl glycosides, followed by diosgenyl glycosides. The total level of four saponins was the highest in fibrous roots and rhizomes, followed by those in the pericarps and arillate seeds, and the lowest in the stems and exarillate seeds. This study can provide data support for the comprehensive development and rational application of non-medicinal parts of PPC.

Traditional use of Paris polyphylla and its active components / 中国中药杂志

This paper reviewed the traditional use of Paris polyphylla and its active components, aiming to provide reference for the development and utilization of this plant. It was found that P. polyphylla has been used as a medicinal plant by eight ethnic minorities. A total of 62 experiential effective recipes, including 29 simple recipes and 33 compound recipes, were analyzed for their indications, traditional processing methods, medicinal compatibilities, and administration doses. The top three in the eight ethnic minorities sorted by the quantity of folk recipes were the Yi nationality(18), Naxi nationality(13) and Bai nationality(12). P. polyphylla has been widely employed for the treatment of nine categories of diseases, especially the dermatologic diseases, trauma, and toxicosis currently. The collating of material basis for its traditional functions revealed 26 active components, among which 19 were steroidal saponins capable of resisting cancer, furuncles, carbuncles, abscesses, bacteria, inflammation and stopping bleeding. This study preliminarily proved the efficacy of P. polyphylla in treating cancer and respiratory system, digestive system, and genitourinary system diseases, which has provided clues for related basic research of P. polyphylla and development of new preparations.

Chemical constituents of steroidal saponins in rhizome of Paris polyphylla var. yunnanensis cultured in vitro / 中国中药杂志

In this study, liquid chromatography-mass spectrometry(LC-MS) and high performance liquid chromatography(HPLC) were employed for qualitative and quantitative analysis of the steroidal saponins in rhizomes of Paris polyphylla var. yunnanensis from three different habitats cultured in vitro, in an attempt to explore whether the rhizomes of the medicinal herb cultured in vitro can synthesize the steroidal saponins, including polyphyllinsⅠ, Ⅱ, and Ⅶ, the quality markers specified in Chinese Pharmacopoeia(2020 edition). A total of 20 steroidal saponins were identified in the rhizomes from Changxin, Yunlong(S1), Fengyi, Dali(S2), and Niujie, Eryuan(S3): parisyunnanoside A and parisyunnanoside D or E, proto-polyphyllin Ⅱ, polyphyllins G and H, polyphyllinsⅠ, Ⅱ, Ⅴ, Ⅵ, and Ⅶ, dioscin, gracillin, prosapogenin A, Tg, isomer of Th, saponin Th, reclinatoside, proto-pairs D, pseudoproto-dioscin, and 23-O-glc-(23S,25R)-spirost-5-en-3β,23α,27-triol-3-O-rha-(1→2)-[ara(1→4)]-glc or 27-O-glc-(23S,25R)-spirost-5-en-3β,27α-diol-3-O-rha-(1→2)-[ara(1→4)]-glc. Among them, polyphyllinsⅠ, Ⅱ, and Ⅶ were detected in the rhizomes from S1, with the mass fraction of 0.109 1%, 0.165 2%, and 0.051 03%, respectively(total 0.325 3%). Polyphyllins Ⅱ and Ⅶ were identified in the rhizomes from S2 with the respective mass fraction of 0.192 2% and 0.074 23% and total content of 0.266 5%. Moreover, polyphyllins Ⅱ and Ⅶ were also found in the rhizomes from S3, which had the mass fraction of 0.207 7% and 0.186 9%, separately, with the total content of 0.394 6%. Thus, steroidal saponins, including the quality makers polyphyllins Ⅰ, Ⅱ, and Ⅶ recorded in Chinese Pharmacopoeia(2020 edition) can be synthesized in rhizomes of Paris polyphylla var. yunnanensis cultured in vitro, but their total content fails to meet the standard(0.60% in Chinese Pharmacopoeia). Therefore, in vitro culture of the Paris polyphylla var. yunnanensis is feasible, but the culture conditions need to be further improved.

Phylogeography of Paris poliphylla var. yunnanensis based on chloroplast gene trnL-trnF sequences / 中国中药杂志

Phylogeography is a research hotspot in the field of the genetic diversity and core germplasm construction of endangered rare plants. Paris polyphylla var. yunnanensis is a rare plant species mainly distributed in China. Wild individuals have been overexploited for the last few decades because of increasing demand for such medicines. Therefore, it is great significance to study the phylogeography of P. poliphylla var. yunnanensis based on chloroplast gene trnL-trnF sequences. In this study, chloroplast genes trnL-trnF were used in the phylogeography analysis of 15 wild and 17 cultivated populations of P. polyphylla var. yunnanensis. This study revealed that based on the results of neutrality tests and mismatch analysis, the rapid expansion of wild population has not been detected in P. polyphylla var. yunnanensis. After aligning and sorting the obtained cpDNA sequences, a total of 15 haplotypes were detected in all 32 populations. One haplotype was unique to the wild population, and 5 haplotypes were unique to the cultivated population. It can be seen that the haplotype richness of cultivated population was higher than that of wild population. The wild populations of P. polyphylla var. yunnanensis were divided into two groups according to evolutionary relationship of haplotypes and distribution map of haplotypes. The haplotype of branch Ⅰ was mainly distributed in Guizhou, and the haplotype of branch Ⅱ was located in Yunnan and Huidong, Sichuan. Therefore, it's speculated that Guizhou and the west Yunnan region may be glacial refuge in the evolutionary history of wild populations of P. polyphylla var. yunnanensis, and in order to protect the wild resources more effectively, wild populations of P. polyphylla var. yunnanensis in these two areas should be included in the protection zone.

Screening and identification of potassium-dissolving bacteria from different rhizosphere soil of Paris polyphylla var. yunnanensis / 中国中药杂志

The study aiming at exploring the potassium-dissolving capacity of rhizosphere potassium-dissolving bacteria from diffe-rent sources and screen the strains with high potassium-dissolving ability, so as to lay a theoretical foundation for cultivation and quality improvement of Paris polyphylla var. yunnanensis sources. The rhizosphere soil of 10 wild and transplanted species from Yunnan, Sichuan and Guizhou provinces was used as the research object. Potassium-dissolving bacteria were isolated and purified, and their potassium-dissolving capacity was determined by flame spectrophotometry, and identified by physiological, biochemical and molecular biological methods. Twenty-six potassium-dissolving bacteria were purified and 13 were obtained from wild and transplanted strains respectively. It was found through the determination of potassium-dissolving capacity that the potassium-dissolving capacity of 26 strains was significantly different, and the mass concentration of K~+ in the fermentation broth were 1.04-2.75 mg·L~(-1), the mcentration of potassium were 0.01-1.82 mg·L~(-1). The strains were identified as Bacillus, Agrobacterium rhizome and Staphylococcus by physiological, biochemical and 16 S rDNA molecular methods, among them Bacillus amylolyticus(4 strains) was the dominant bacterium of Bacillus. The physiology and biochemistry of rhizosphere potassium-dissolving bacteria in P. polyphylla var. yunnanensis rhizosphere were diffe-rent, and the living environment were different, so the potassium-dissolving capacity also changed. Strain Y4-1 with the highest potassium decomposability was Bacillus amylolytic with a potassium increase of 1.82 mg·L~(-1). The potassium-dissolving ability and the distribution of potassium-dissolving bacteria were different in various habitats. The screening of potassium-dissolving bacteria provided a new strain for the preparation of microbial fertilizer. It is expected that B. amyloidococcus Y4-1 can be used as an ideal strain to cultivate mycorrhizal seedlings of P. polyphylla var. yunnanensis.

Cloning and Prokaryotic Expression of Glycosyltransferase Genes from Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

Objective:To clone the full-length glycosyltransferase genes （<italic>PpUGT</italic>1，<italic>PpUGT</italic>7） related to saponins biosynthesis in <italic>Paris polyphylla</italic> var. <italic>yunnanensis</italic>，and perform bioinformatics analysis，relative expression analysis and prokaryotic expression analysis. Method:Total RNA was isolated from <italic>P. polyphylla </italic>var. <italic>yunnanensis </italic>with use of the Eastep^® Super Total RNA Extraction Kit and converted to cDNA. Specific primers were designed according to the transcriptome data to clone the full-length gene. Relevant software was then used for bioinformatic analysis of the protein sequences. The relative gene expression levels were detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and the prokaryotic expression vectors were built to heterologously express recombinant protein in <italic>Escherichia coli.</italic> Result:The open reading frame （ORF） of <italic>PpUGT</italic>1 was 1 827 bp，encoding 608 amino acids，and was predicted as a steroid glycosyltransferase；the ORF of <italic>PpUGT</italic>7 was 1 380 bp，encoding 459 amino acids，and was predicted as a triterpenoid glycosyltransferase. The calculated relative molecular mass of two proteins were 67.6 kDa and 51.3 kDa respectively，and both of them were hydrophilic proteins，no transmembrane domain，no signal peptides，both showing high similarity and conservativeness with homologous sequences. The results of Real-time PCR showed that the expression level of <italic>PpUGT</italic>1 was root>leaf>flower>stem；the expression level of <italic>PpUGT</italic>7 was stem>leaf>flower>root. In addition，PpUGTs proteins were expressed in <italic>E. coli</italic>. in a soluble form. Conclusion:The genes of <italic>PpUGT</italic>1 and <italic>PpUGT</italic>7 were cloned successfully. Real-time PCR showed the genes were expressed differently in different plant organs， and their recombinant proteins were successfully expressed in <italic>Escherichia coli</italic>. This study lays a foundation for functional characterization of PpUGTs and analysis of the biosynthesis pathway of saponins in <italic>Paris polyphylla </italic>var. <italic>yunnanensis</italic>.

Effect of Different Combinations of Arbuscular Mycorrhiza on Paris polyphylla var. yunnanensis and Chemical Components / 中国实验方剂学杂志

Objective:To study the effects of different arbuscular mycorrhizal（AM） fungi combinations on the growth and quality of<italic> Paris polyphylla</italic> var. <italic>yunnanensis</italic> seedlings，in order to provide reference for the cultivation of high quality <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>. Method:Through the room temperature pot inoculation test method，nine groups of different AM fungi combinations were inoculated into sterilized soil，and the control group was not inoculated. The effects of different AM fungi combinations on root infection rate，photosynthetic parameters，physiological indexes and chemical components of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> seedlings were observed. Result:The spore density in rhizosphere soil，the infection rate and intensity of AM fungi in roots，and the activities of succinate dehydrogenase and alkaline phosphatase in roots of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> were significantly increased by inoculating the combination of exogenous AM fungi. Compared with the control group，the contents of chlorophyll，malondialdehyde，soluble sugar，soluble protein content and the activities of antioxidant enzymes in leaves of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> were increased in different AM fungi treatment groups. The content of total saponin increased from T1 to T2，decreased rapidly from T2 to T4，and increased at T5. Conclusion:Different mixed AM fungus can form a good symbiotic relationship with the roots of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>，improve the mycorrhizal vitality，enhance the resistance of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> to adverse environment，promote the growth and development of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> and improve the quality of rhizome. According to the comprehensive inoculation effect，the combination of S5，S8 and S9 AM fungus has the best effect，which provides reference value for the application of mycorrhizal biotechnology of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>.

Influences of Natural Fibrous Root Decay on Growth and Active Component Content of Paris polyphylla var. yunnanensis / 中国实验方剂学杂志

Objective:To explore the effect of natural decay of exogenously added fibrous roots on the growth and development of <italic>Paris polyphylla</italic> var. <italic>yunnanensis </italic>and its medicinal quality. Method:The effects of natural decay of fibrous roots at different amounts on mycorrhizal infection rate， physiological and biochemical indexes， and saponin contents of <italic>P. polyphylla</italic> var. <italic>yunnanensis </italic>were studied in pot culture experiments at room temperature. Result:The results showed that the infection rate of arbuscular mycorrhizal （AM） fungi in the root of <italic>P. polyphylla</italic> var. <italic>yunnanensis </italic>was not significantly affected by different fibrous root treatments， but there were significant differences in infection intensity. The photosynthetic pigment content in the leaves declined significantly with the increase in fibrous root amount， and the total chlorophyll was decreased by 78.7% at most. The contents of soluble protein， soluble sugar and malondialdehyde in the leaves of <italic>P. polyphylla</italic> var. <italic>yunnanensis </italic>showed an overall upward trend. The activities of the three protective enzymes varied. The peroxidase and malondialdehyde were reduced by 181.6% and 200.0% at most. In the root system of <italic>P. polyphylla </italic>var. <italic>yunnanensis</italic>， the contents of the above-mentioned six components decreased to varying degrees， with the largest reductions of peroxidase and malondialdehyde reaching 44.6% and 69.7%. Different fibrous root treatments resulted in a decrease in active component content of <italic>P. polyphylla </italic>var. <italic>yunnanensis</italic>. The total content of the four saponins was decreased by 58.9% at most， and the total saponin content by 46.9%. Conclusion:The natural decay of fibrous roots affects the soil microbial environment of root system， reduces the photosynthetic pigment content in leaves， and destroys the stability of cells， thus interfering with the growth and development of <italic>P. polyphylla </italic>var. <italic>yunnanensis</italic>， reducing its medicinal components， and causing continuous cropping obstacles.

Paris saponin VII, a direct activator of AMPK, induces autophagy and exhibits therapeutic potential in non-small-cell lung cancer / 中国天然药物

Paris saponin VII (PSVII), a bioactive constituent extracted from Trillium tschonoskii Maxim., is cytotoxic to several cancer types. This study was designed to explore whether PSVII prevents non-small-cell lung cancer (NSCLC) proliferation and to investigate its molecular target. AMP-activated protein kinase (AMPK) has been implicated in the activation of autophagy in distinct tissues. In cultured human NSCLC cell lines, PSVII induces autophagy by activating AMPK and inhibiting mTOR signaling. Furthermore, PSVII-induced autophagy activation was reversed by the AMPK inhibitor compound C. Computational docking analysis showed that PSVII directly interacted with the allosteric drug and metabolite site of AMPK to stabilize its activation. Microscale thermophoresis assay and drug affinity responsive target stability assay further confirmed the high affinity between PSVII and AMPK. In summary, PSVII acts as a direct AMPK activator to induce cell autophagy, which inhibits the growth of NSCLC cells. In the future, PSVII therapy should be applied to treat patients with NSCLC.

Isolation and identification of phosphatolytic bacteria in Paris polyphylla var. yunnanensis / 中国中药杂志

The wild resources of Paris polyphylla var. yunnanensis, a secondary endangered medicinal plant, are severely scarce. Introduction and cultivation can alleviate market demand. To screen phosphatolytic bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis and provide data support for the development of high-efficiency microbial fertilizer, in this study, the dilution plate coating method was used to isolate and screen the phosphorus solubilizing bacteria with the ability of mineralizing organic phosphorus from the rhizosphere soil of wild and transplanted varieties of P. polyphylla var. yunnanensis in 10 different locations in Yunnan, Sichuan and Guizhou. After separation and purification, the phosphatolytic capacity was analyzed by qualitative and quantitative analysis. Combined with physiological and biochemical experiments, the strains were identified using 16 S rDNA sequencing analysis. Forty one strains were selected from the rhizosphere soil of P. polyphylla var. yunnanensis from 10 different habitats. Among them, 21 strains were obtained from the rhizosphere soil of the wild variety P. polyphylla var. yunnanensis and 20 strains were obtained from the rhizosphere soil of the transplanted variety. And significance analysis found that 41 organophosphate solubilizing strains had significant differences in their ability to solubilize phosphorus. The amount of phosphate solubilizing was 0.08-67.61 mg·L~(-1), the pH value was between 4.27 and 6.82. The phosphatolytic amount of strain Y3-5 was 67.61 mg·L~(-1), and the phosphorus increase amount was 57.57 mg·L~(-1). All 41 strains were identified as Gram-positive Bacillus. Combining physiological characteristic and phylogenetic trees, Bacillus mobilis Y3-5 was finally selected as the candidate rhizosphere phosphatolytic bacteria of P. polyphylla var. yunnanensis. The distribution of phosphorus solubilizing bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis was different, and there were significant diffe-rences in phosphorus solubility. Organophosphate-dissolving strain Y3-5 is expected to be a candidate strain of P. polyphylla var. yunnanensis microbial fertilizer.

A Comparative Study on the Fingerprint of Paris polyphylla var. Polyphylla and Paris polyphylla var. yunnanensis from Different Origins in Dali and Seven Main Steroidal Saponins by HPLC / 中国药学杂志

OBJECTIVE: To study the fingerprints of rhizomes of Paris polyphylla Smith var. polyphylla and Paris polyphylla Smith var. yunnanensis (Franch.) Hand. -Mazz from different origins in Dali and the differences of seven main steroidal saponins. METHODS: The fingerprints of rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali were established by HPLC. The similarity of fingerprints and seven main steroidal saponins were compared and analyzed. RESULTS: There were 14 common peaks in the fingerprints of rhizomes of Paris polyphylla var. polyphylla from different origins in Dali, and the similarity of the fingerprints of the samples from different habitats except Jinhua Weishan and Longjie Weishan was greater than 0.9. There were 13 common peaks in the fingerprints of rhizomes of Paris polyphylla var. yunnanensis from different origins in Dali, and the similarity of the samples from different origins was greater than 0.92. There were 11 common peaks in the fingerprints of rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali, and the similarity of the fingerprints was very low, between 0.057 and 0.225. Polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ are detected in the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali. The average peak areas of polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ in Paris polyphylla var. yunnanensis from different origins in Dali were higher than those of Paris polyphylla var. polyphylla. Polyphyllin H was detected in the rhizomes of Paris polyphylla var. yunnanensis from Leqiu Nanjian, Jinhua Weishan, Fengyu Eryuan, Dengchuan Eryuan, Hongyan Midu, Xiyi Heqing and the rhizomes of Paris polyphylla var. polyphylla from Wanqiao Dali, Fengyi Dali, Xiyi Heqing, Hongyan Midu and Leqiu Nanjian. Polyphyllin Ⅲ was detected in the rhizomes of Paris polyphylla var. polyphylla from the origins except Longjie Weishan. Polyphyllin Ⅴ was also detected in the rhizomes of Paris polyphylla var. polyphylla in Fengyu Eryuan and Xiyi Heqing. CONCLUSION: Polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅶ are detected in the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis from different origins in Dali. The similarity of the HPLC fingerprints of the rhizomes of Paris polyphylla var. polyphylla and Paris polyphylla var. yunnanensis is very low, and there are great differences in seven main steroidal saponins. The contents of polyphyllin Ⅰ, polyphyllin Ⅱ, polyphyllin Ⅵ and polyphyllin Ⅴ in Paris polyphylla var. yunnanensis are higher than those in Paris var. polyphylla.