Rapid identification of Paris polyphylla var. yunnanensis and its closely relative species by infrared spectroscopy combined with morphological characteristics / 中草药

Objective: To explore morphology and infrared spectrum identification evidence of Paris polyphylla var. yunnanensis and its closely relative species, and further analyze their genetic relationship to provide basis for the development and utilization of medicinal plant resources of genus Paris. Methods: The morphology and infrared spectrum of P. polyphylla var. yunnanensis and its closely relative species were studied systematically and compared with each other. The original infrared spectra data were pretreated by automatic baseline correction, automatic smoothing, ordinate normalization, and second derivative, and analyzed by principal component analysis (PCA), partial least squares discrimination analysis (PLS-DA), and hierarchical cluster analysis (HCA). Results: There were typical characteristics of P. polyphylla var. yunnanensis and its closely relative species, such as plant size, leaves, veins, sepals, petals, flower pavilions and stamens, which could provide morphological identification evidence. There were obviously differences of the second derivative in 3 000-2 000 cm-1 and the fingerprint in 1 800-500 cm-1 in the six types of genus Paris. Both HCA and PLS-DA could better distinguish Paris polyphylla var. yunnanensis and its closely relative species, which could provide an infrared spectral identified evidence. The Results: of HCA showed that P. polyphylla var. yunnanensis, P. polyphylla var. chinensis, P. polyphylla, P. polyphylla var. stenophylla and P. polyphylla var. nana were relatively close. Conclusion: Fourier transform infrared spectroscopy (FTIR) combined with original plant morphological identification can quickly identify P. polyphylla var. yunnanensis and its closely relative species, which will provide a scientific basis for the cultivation, clinical application, and resource development of genus Paris.

Effects of combined application of N, P and K on yield and active ingredient content of rhizome of Paris polyphylla var. chinensis / 中草药

Objective: To research the effects of different gradient levels of N, P, and K of rhizome of Paris polyphylla var. chinensis yield and quality, determine the optimal fertilization scheme and provide a scientific basis for rational fertilization. Methods: The three-factor five-level quadratic general rotation combination design was adopted for field experiment, correlation measurement and data analysis were carried out, and the effects of combined application of N, P, and K on weight gain rate, total saponin content, ash, moisture and extracts were investigated. Results: The established regression equations reached significant levels. When applying 41.66-52.60 kg/hm2 of N, 181.38-244.38 kg/hm2 of P, 24.16-35.20 kg/hm2 of K, the rhizome weight gain rate of P. polyphylla var. chinensis was greater than 81.72%; When applying 45.48-53.83 kg/hm2 of N, 179.98-236.83 kg/hm2 of P, 29.80-39.95 kg/hm2 of K, the total saponin content in the rhizome of P. polyphylla var. chinensis was greater than 11.09%; When applying 28.01-37.79 kg/hm2 of N, 127.18-209.18 kg/hm2 of P, 25.27-34.09 kg/hm2 of K, the extract content of the rhizome of P. polyphylla var. chinensis was more than 14.31%. Conclusion: The optimum fertilization amount on the rhizome of P. polyphylla var. chinensis was as follow: 37.79-45.48 kg/hm2 of N, 181.38-209.18 kg/hm2 of P, and 29.80-34.09 kg/hm2 of K.

Effect of dynamic distribution of steroid saponins from Paris polyphylla var. chinensis on medical material quality / 中草药

Objective: To obtain the optimum cultivation period and best harvesting month of P. polyphylla var. chinensis by studying on change regularity of biomass accumulation rate and steroid saponins from P. polyphylla var. chinensis with the changes of its harvesting time and medicinal parts. Methods: A high performance liquid chromatography-evaporative light-scattering detector (HPLC-ELSD) method was established to characterize content of six steroid saponins (Paris saponins I, II, VI, VII, V, and H) from P. polyphylla var. chinensis and its dynamic change regularity was detected; Drying rate was measured by oven-dry method to indicate biomass accumulation rate. Results: Drying rate increased with the planting ages and reached the highest level in the eighth year. There was a waving trend between drying rate and harvesting time and drying rate reached crest in January and June; The main steroid saponins of P. polyphylla var. chinensis were Paris saponin VII and Paris saponin H. The content of total saponins varied by `V' shaped with the increases of planting-year and reached the highest in the second and eighth year, and it changed by waving shape with the change of harvesting month and reached the crest in March and November; There was a distinct difference of total saponins among different parts of P. polyphylla var. chinensis: stems and leaves > rhizome-forepart > rhizome-middle-part ≈ rhizome-end-part. The distribution of paris saponins VII decreased obviously from stems to rhizome-end-part while paris saponins H increased gradually. Conclusion: There were remarkable correlations among steroid saponins accumulation amounts and planting ages, harvesting month, and medicinal parts. The optimum cultivation period is eight years and best harvesting month is in November; The quality of medicinal materials would decline if only rhizome-end-part to be used; Paris saponin VII maybe a relative factor for germination and growth of P. polyphylla var. chinensis; Stems of P. polyphylla var. chinensis can be an important resource to extract steroid saponins, which have a profound significance to broaden resources of medicinal plants and improve resources utilization efficiency.

Cultivable bacterial diversity of Paris polyphylla var. chinensis with 16 S rRNA sequence analysis / 中草药

Objective To analyze the bacterial diversity in stem rot and healthy plants of Paris polyphylla var. chinensis. Methods Bacterial strains were isolated from rhizomes, stems and leaves of the diseased and healthy plants of P. polyphylla var. chinensis using beef extract-peptone medium. Using 16 S rRNA universal primers 27F/1492R for PCR amplification, combined with DNA sequencing technology to preliminary identification the bacterial strains. Results The bacteria could be divided into 23 microbial species, belonging to 11 genera, isolated from healthy and diseased plants of P. polyphylla var. chinensis. Eleven endophytic bacteria, belonging to 4 genera were contained from healthy plants, in which there were 9, 10, and 5 species isolated from rhizomes, stems and leaves, respectively. Fourteen endophytic bacteria, belonging to 10 genera were contained from diseased plant, in which there were 11, 8, and 3 species isolated from rhizomes, stems, and leaves, respectively. The content of endophytic bacteria in rhizome of healthy plant was the highest, reached up to 2.999 × 105 cfu/g, while that in leaf was the lowest with 7.32 × 104 cfu/g. The quantities of Bacillus species in rhizome (73.3%), stem (67.1%), and leaf (81.8%) of healthy plant were the highest groups, Pseudomonas species in rhizome (35.6%), stem (50.3%) and leaf (60.5%) of diseased plants were the highest groups. Shannon-Wiener index and evenness index of healthy plant of P. polyphylla var. chinensis were higher than that of diseased plants. Conclusion The dominant group in the healthy plant of P. polyphylla var. chinensis was Bacillus, while the dominant groups in the diseased plant of P. polyphylla var. chinensis was Pseudomonas. The population diversity of the cultivable bacteria in healthy plant of P. polyphylla var. chinensis was more abundant than that of diseased plant.

Physicochemical changes of Paris polyphylla var. chinensis seed during different stages of germination / 中草药

Objective To investigate the regulation mechanism of enzymes and endogenous hormones of different stages of Paris polyphylla Smith var. chinensis seed germination, aiming to provide scientific basis for taking effective treatment measures during the different stages of seed dormancy and germination to increase seedling emergence rate. Methods The morphological differences of different stages of P. polyphylla Smith var. chinensis seeds germination after treatment with variable temperature stratification were observed and measured. Activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), malic acid deoxy enzyme (MDH) and 6-phosphate dehydrogenase (G-6-PDH) were determined by routine physiological and biochemical methods. Levels of 6 endogenous hormones, named indole acetic acid (IAA), gibberellin (GA3), abscisic acid (ABA), salicylic acid (SA), Trans-Zeatin (TZ), and jasmonic acid (JA) were quantitatively determined by mass spectrometry. The correlation among enzyme activity, endogenous hormone content and embryo rate during seed germination was studied. Results In the stages of non-germinating, epicotyl dormancy and upcoming emerging of P. polyphylla Smith var. chinensis seeds, seed weight and embryo rate increased gradually; the activities of SOD and POD increased gradually, while activities of CAT and MDH decreased gradually. The G-6-PDH activity levels showed two peaks and the levels of non-germinating stage and stage before epicotyl elongation after germination were obviously higher than those of other stages. The contents of IAA, GA3, ABA and JA increased gradually, and the contents of IAA and JA were significantly higher in the upcoming emerging stage than those of other stages (P < 0.01). The contents of TZ and SA increased first and then decreased. TZ levels of cotyledon differentiation stage and endosperm maturation stage were obviously higher than those of other stages (P < 0.01). SA level was highest in the early stage of emerge, and the levels of SA in germinating radicle just breaking through the seed coat stage and endosperm formation stage were second. The change of CAT enzyme activity was negatively correlated with embryo rate, and the content of IAA and JA was positively correlated with embryo rate. Conclusion CAT could inhibit the growth of embryo, and IAA and JA had positive effects on the growth of embryo.

Evaluation of saponins in Paris Polyphylla var.chinensis from twenty-one growing areas / 中成药

AIM To evaluate saponins in Paris Polyphylla Smith var.chinensis (Franch.) Hara from twentyone growing areas.METHODS HPLC was adopted in the content determination of pennogenins { pennogenin-3-O-β-D-glu (1→3) [α-L-rha (1→2)]-β-D-glu (PGGR),polyphyllinsⅥ,Ⅶ,H} and dioscins (gracillin,polyphyllins Ⅰ,Ⅱ,Ⅴ),then SPSS 22.0 software was used for principal component analysis and cluster analysis.RESULTS Pennogenin was the main kind of Paris Polyphylla var.chinensis saponins,and PGGR,polyphyllins Ⅰ,Ⅴ,Ⅶ,H were the main effective constituents of this medicinal material.The comprehension scores of seven growing areas were more than zero,which was in sequence of Bazhong ＞ Qingchuan ＞ Yanjin ＞ Tongjiang ＞ Nanchuan ＞ Chongzhou ＞ Hongya.Twenty-one batches of samples were divided into four types,which was similar to cluster analysis results.CONCLUSION Bazhong,Qingchuan,Chongzhou,Hongya,Tongjiang in Sichuan,Yanjin in Yunnan,and Nanchuan in Chongqing are suitable for cultivating Paris Polyphylla var.chinensis,which can guarantee the stability of saponins.

Optimization for Extracting Polysaccharide from Paris polyphylla Smith var. chinensis (Franch.) Hara in Shaan'xi by Response Surface Method / 医药导报

Objective To select the optimum extracting method of polysaccharide from Paris polyphylla Smith var. chinensis(Franch. )Hara in Shaan'xi. Methods The optimum extraction technology and condition were determined by using response surface methodology and single factor experiment,with the extraction ratio of polysaccharides from Paris polyphylla Smith var. chinensis( Franch. ) Hara in Shaan'xi as indicator. Results The optimum extraction condition for the polysaccharides was as follows:extracting at 92 ℃ for 2. 6 h and with the liquid to solid ratio of 15 ( mLg). The average extraction yield of polysaccharide was 3. 22%under the optimum extraction condition. Conclusion The method is suitable for the extraction and determination of polysaccharides from Paris polyphylla Smith var. chinensis( Franch. ) Hara in Shaan'xi.