Application of Metabolomics Approach to Study on Chemical Constituents in Cultivated and Wild Pinellia ternate (Thunb.) Breit. and Typhonium flagelliforme (Lodd.) Blume Based on UPLC-ESI-HRMS^n

[Objectives] The aim was to study chemical components of cultivated and wild Pinellia ternate(Thunb.) Breit and Typhonium flagelliforme(Lodd.) Blume by metabolomics technology based on UPLC-ESI-HRMS^n. [Methods] Chemical components of 27 samples of cultivated and wild P. ternate and T. flagelliforme were detected by using UPLC-ESI-HRMS^ntechnology. Differential markers between P. ternate and T. flagelliforme as well as cultivated and wild P. ternate were screened and identified by principal component analysis(PCA) and orthogonal partial least square-discriminate analysis(OPLS-DA). [Results]12 components of P. ternate(cultivated and wild variety) and T.flagelliforme and three components of cultivated and wild P. ternate were screened and identified by OPLS-DA. N-Acetyl-6-hydroxy-L-tryptophan and N-Phenylacetylglutamic were not detected in P. ternate,and these two components can be used as potential markers of P. ternate and T. flagelliforme. [Conclusions]UPLC-ESI-HRMS^n-based metabolomics technology could be used to quickly identify the chemical constituents of P. ternate and effectively distinguish P. ternate and T. flagelliforme,which has reference significance for the quality control of P. ternate.

Optimization of Conditions for Extracting Luteolin from Ternate Grape Fern Herb by an Orthogonal Experiment

[Objectives] This study was conducted to optimize the extraction process of luteolin, a natural product of Ternate Grape Fern Herb, so as to provide a theoretical research basis for the chemical composition, pharmacological research and preparation development of Ternate Grape Fern Herb. [Methods] The effects of four factors including ultrasonic extraction time, liquid-to-material ratio, ultrasonic extraction temperature and ethanol concentration on extraction yield of luteolin from Ternate Grape Fern Herb were investigated by the ultrasonic-assisted extraction technology, through a single-factor experiment and an orthogonal experiment based on the single-factor experiment, and the experimental data was subjected to variance analysis and SSR analysis of the significance of differences between groups by SPSS20.0 software to select the optimal extraction process conditions. [Results] The optimal extraction process conditions were as follows: ultrasonic extraction time 25 min, liquid-to-material ratio 25∶1 ml/g, ethanol volume fraction 60%, ultrasonic extraction temperature 85 ℃. [Conclusions] The results were verified, and it was determined that the ultrasonic-assisted extraction process established in this study is reasonable and reliable, and can be used for the extraction of luteolin.

Study of dual-directional regulatory effect of Banxia （Pinellia ternata） and Huanglian （Coptis chinensis） drug pair on gastrointestinal movement of mice

Objective： To evaluate the regulatory effects of Banxia （Pinellia ternata, P） and Huanglian （Coptis chinensis, C） drugpair, derived from Banxiaxiexin soup, on the gastrointestinal movement of mice with functional dyspepsia. Methods：Mice were treated with different proportions of the P and C drug pair （1：1, 3：1, and 4：1） for 10 days, and subsequentlyinjected with atropine （ATR） or neostigmine （NEO）. The effects of the different proportions of P and C were evaluatedbased on the alvine advance rate. In addition, we used the same modeling method used in the first experiment andadministered P： C at ratio of 3：1 and at different doses respectively （4.68 g/L, 2.34 g/L, and 1.17 g/L）, and tested levelsof the gastrointestinal hormones, gastrin （GAS）, vasoactive intestinal polypeptide （VIP）, and somatostatin （SS） in thesmall intestinal tissue using an enzyme-linked immunosorbent assay. Results： In the groups of NEO-induced mice, P：Cat ratios of 1：1, 3：1, and 4：1 significantly reduced the alvine advance rate compared with the NEO model group （P =0.003, P = 0.012 and P = 0.021, respectively）. In the groups of ATR-induced mice, only P：1 at ratio of 3：1 significantlyincreased the alvine advance rate compared with the ATR model group （P = 0.007）. After exposure to P： C at ratio of 3：1and at different dose, the GAS level was lower in the low-, medium-, and high-dose NEO groups than that in the NEOmodel group （P = 0.001, P = 0.004, and P = 0.003, respectively）. The VIP levels were higher in the medium-andhigh-dose NEO groups than that in the NEO model group （P = 0.004 and P = 0.002, respectively）. In addition, the SSlevel increased in the NEO medium-dose group compared with that in the NEO model group （P = 0.002）. The GAS levelwas higher in the ATR medium- and high-dose groups than in the ATR model group （P = 0.007 and P = 0.021,respectively）. The VIP level was lower in the ATR low-, medium-, and high-dose than that in the ATR model group （P =0.001, P = 0.001, and P = 0.001, respectively）. Furthermore, the SS level was lower in the ATR medium- and high-dosegroups than that in the ATR model group （P = 0.001 and P = 0.006）. Conclusion： The PC drug pair bidirectionallyadjusted the NEO- and ATR-induced functional dyspepsia in mice by modulating GAS, VIP, and SS levels in theintestine.

基于16S rDNA分析湖北半夏内生菌群结构及代谢功能

为探究湖北半夏(Pinellia ternate)内生细菌的多样性、丰度及菌群功能,随机采集半夏植株样品,提取内生细菌基因组DNA,通过PCR扩增获得16S rDNA V3~V4片段。采用IlluminaMiSeq高通量测序技术分析半夏植株中的内生细菌菌群结构,共获得有效序列39537条和用于物种分类的OTU数681条,包括6门、10纲、23目、29科、45属、26种的细菌。菌落多样性分析表明,湖北半夏内生细菌在门水平上,优势菌群为Proteobacteria,科水平上,Mitochondria和Pseudomonadaceae为优势菌群。功能预测结果表明,湖北半夏内生细菌编码的大多数基因与代谢相关,其次是基因信息处理、环境信息处理。

用于治疗新冠肺炎的中药材半夏道地性保护研究

中药材半夏(Pinellia ternate)是华中道地药材产区优势道地药材品种,为治疗新型冠状病毒肺炎(COVID-19)中医国家新处方清肺排毒汤的21味中药材之一。介绍了半夏的药用价值及其中药材GAP基地、贫困地区生态适宜种植县域;总结了涉及半夏的知识产权资源;从农业知识产权、特色农产品优势区、道地药材生产基地建设等方面探讨了乡村振兴战略与中医药发展战略下中药材半夏道地性保护与高质量发展的策略。

不同采样点息半夏质量对比研究

分别从性状鉴别、粉末显微鉴别、薄层色谱鉴别、水分、总灰分、浸出物检查、总酸含量测定等方面,比较了7个采样点息半夏(Pinellia ternate(Thunb.)Breit of Xixian county)的质量。结果表明,7个采样点息半夏粉末显微特征、薄层色谱一致;但性状、水分、总灰分、浸出物和总酸含量有差异。其种源、繁育方式、野生环境、培育条件对息半夏质量有影响。

FORCING INTERCELLULAR CYTOPLASM AND CHROMATIN MIGRATION BETWEEN THE MERISTEMTIC CELLS OF Pinellia ternate AND Commelina communis BY LB TECH NIQUE AND THE SUBSEQUENT REGENERATION OF HYBRID PLANTS

The interfamilical hybrid plants with color character for genetic marker of Pinellia ternate and Commelina communis have been obtained successfully by the LB technique——forcing intercellular cytoplasm and chromatin migration between cells of different plants by artificial motive force in our lab. The preliminary results are as follows.

The late Pliocene species with ternate venation of Lauraceae from Tuantian flora, Tengchong, Yunnan Province, China

A number of fossil leaves of the Lauraceae were collected from Tuantian flora of the late Pliocene strata in Yunnan, China. They have a whole leaf shape and well-preserved cuticles. In this paper, features of the architecture and epidermis of three species of the Lauraceae family with ternate venation are reported. Results of the comparison between the venation characteristics of Lauraceae fossils from four fossil floras in the world and those of extant Lauraceae in China suggest that the proportion of Lauraceae species with ternate venation is increasing. The proportion has risen from 10% in the Eocene to 12.5% in the late Pliocene in China, whereas the proportion in the modern era is 21%. Moreover, the proportion of Australian floras with ternate venation has rised from 8.3% to 10% between the Eocene and the Miocene. Because of the effect of the global drought during the Cenozoic era, we suggested that species with ternate venation in Lauraceae have become increasingly prevalent between the Eocene era and the present day in China. Data on Nerriga flora and Maarlake flora since the Eocene era in the Southern Hemisphere show the same trend. While features of the leaf architecture and epidermis of Lindera acuminatissima differ in details. They all have similar identification characteristics with regard to basal eucamptodromous venation, interior 2° present, minor 2° simple brochidodromous, anticlinal walls, no surface ornamentation, lip-like guard cells, outer stomatal ledge plain or dimness, and paracytic stomatal aperture. These differences are also present in extant species. Compared with Lindera tonkinensis, the nearest living relative(NLR) of L. acuminatissima shows that these differences are relevant to the growth, position, and development of leaves. Leaf-type classification statistics of L. acuminatissima indicate that when all fossil leaves are taken into consideration, A-type leaves account for 24%, B-type leaves make up 56%, and C-typy leaves account for 20%. Comparing the above result with that of NLRs, A-type leaves are generally immature spires and cannot be preserved easily, B-type leaves are regular mature leaves and fossilize more easil, and C-type leaves are usually old leaves occurring at the bottom, broken, ill and consumed by worms.

外源添加物对半夏愈伤组织次生代谢的调控

以半夏[Pinellia ternate(Thunb.)Breit]愈伤组织为材料,研究不同外源添加物对半夏愈伤组织内GOGAT、GDH、GOT、PAL活性和总生物碱、鸟苷和腺苷含量的影响。结果表明,不同浓度的外源添加物SNP、ASA、Phe、Asp单一或两两复合添加,均能提高半夏愈伤组织中GOGAT、GDH、GOT、PAL的活性,增加总生物碱和鸟苷的含量。其中,1 mg/L ASA和150 mg/L Asp复合处理诱导GOGAT、GDH、GOT、PAL活性提高和可溶性蛋白含量增加的效果最好,此时半夏愈伤组织中总生物碱及鸟苷含量分别比对照增加131.0%、146.7%。适宜浓度的SNP、ASA、Phe、Asp可通过提高半夏愈伤组织中GOGAT、GDH、GOT、PAL活性加速无机氮的同化及转化效率,促进总生物碱、鸟苷和腺苷的合成与积累。

Simultaneous determination of 6 organic acids,3 nucleosides,and ephedrine in Pinellia ternata by HPLC

Pinellia ternata has a long history of medicinal use in China, Japan, and Korea. Herein, we present a sensitive, selective, and robust HPLC method for the simultaneous determination of 6 organic acids （oxalic acid, malic acid, citric acid, formic acid, acetic acid, and succinic acid）, 3 nucleosides （uridine, guanosine hydrate, and adenosine）, and ephedrine in P. ternata. The Rhizoma pinelliae samples were collected from different parts of Southwest China, and the 10 constituents were detected at a wavelength of 210 nm. Chromatographic separation was achieved on a Boston Green ODS reversed-phase column along with a gradient mobile phase of methanol and phosphate buffer （pH 2.3, 10 mmol/L, v/v） at a flow rate of 1.0 mL/min. The range of recovery was between 90.8% and 101.9%, and the calibration curves showed high linearity. The relative standard deviations of the new method were less than 3.0% for intra- and inter-day analyses. All samples were stable for at least 48 h. The abundance of the 10 compounds were dependent on their specific locations. The new HPLC method is a powerful technique for the quality control ofP. Ternata.