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ObjectiveTo screen and validate key enzyme genes affecting the polysaccharide content in different Polygonatum species and perform in-depth amino acid sequence analysis by transcriptomic analysis of P. zanlanscianense， P. kingianum， and P. cyrtonema rhizomes to enrich the transcriptome data of Polygonatum plants and provide references for polysaccharide biosynthesis mechanism and genetic improvement. MethodThe Polygonatum transcriptome was sequenced and analyzed using the Illumina NovaSeq high-throughput sequencing platform， and the differences in the transcriptomes of the three Polygonatum species were compared and according to the annotations of Nr， Gene Ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） databases. The key enzymes in the polysaccharide metabolism pathway were screened， and the expression of key enzyme genes was clustered and correlated with the polysaccharide content. Finally， Real-time polymerase chain reaction （Real-time PCR） was performed to validate the eight key enzyme genes， and the key genes of polysaccharide biosynthesis were further screened for homologous gene sequence analysis in combination with sequencing results， followed by constructing phylogenetic trees， predicting motifs， conserved structural domains， protein sequence isoelectric points， and molecular weights， and constructing 3D protein structures by using homology modeling method. ResultThe annotation of the Nr database revealed that three Polygonatum species had the highest gene homology with Asparagus officinalis. GO database annotation results showed that three Polygonatum species differed significantly in binding， catalytic activity， metabolic processes， and cellular components， while the KEGG pathway annotation results indicated that three Polygonatum species differed significantly in the starch and sucrose metabolic pathway and galactose metabolic pathway. According to clustering analysis， correlation analysis， Real-time PCR， expression profiles， and structural and functional predictions of amino acid sequences， the key enzyme significantly affecting the polysaccharide content in different Polygonatum species was inferred to be β-fructofuranosidase （sacA）. ConclusionSacA may be the main influencing factor for the difference in polysaccharide content of Polygonatum， and is also an important reason why Polygonatum polysaccharides are mainly fructans.
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OBJECTIVE：To establish the method for the content determination of inorganic elements in Cyperus rotundus ，and to compare the contents of 14 kinds of inorganic elements in C. rotundus from different producing areas ，and to provide theoretical basis for its quality control and high quality resources development . METHODS ：The samples were processed by microwave digestion，and ICP-MS method was used to determine the contents of Na ，Mg，K，Ca，Mn，Fe，Ni，Cu，Zn，As，Se，Sr，Cd and Pb. SPSS 23.0 software were used for principal component analysis （PCA）and cluster analysis. RESULTS ：The average contents of above 14 kinds of inorganic elements in C. rotundus from different producing areas were 168.62，753.71， 6 938.33，24.31，14.69，197.77，0.60，2.43，26.89，0.21，0.06，5.64，0.05，0.32 mg/kg，respectively. The results of PCA showed that the cumulative variance contribution rate of the first four principal components was 86.203％，which could reflect most of the information of the original data. C. rotundus from Shandong ，Jiangxi，Shanxi，Hubei and Yunnan ranked the top five places in terms of comprehensive score of inorganic element contents. The results of cluster analysis showed that the samples from 9 producing areas were clustered into 5 categories，showing the characteristics of clustering by producing area. From the perspective of inorganic elements ，the quality of C. rotundus from East China ，Central China ，North China and Southwest China was better than that from South China. CONCLUSIONS ：Essential trace elements like Na ，Mg，K，Ca，Mn，Fe，Cu，Zn，Sr are rich in C. rotundus，and there are small amounts of Ni ，As，Se，Cd，Pb elements in it. The contents difference of inorganic elements in C. rotundus from different origins may related to the geographical area it belongs to.
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OBJECTIVE： To estab lish the method for simultaneous determination of the contents of cyperotundone ， nootkatone，α-cyperone and aristolone in the volatile oil of Cyperus rotundus ，compare the content differences of 4 components in C. rotundus samples from different origins ，and to provide reference for germplasm screening ，development and utilization of the medicinal material. METHODS ：The volatile oil was extracted from 46 batches of C. rotundus from 12 origins. The contents of cypermenone，nootkatone，α-cyperone and aristolone in volatile oil were determined by HPLC. The determination was performed on Kromasil C 18 column with mobile phase consisted of methanol-water （68∶32，V/V）at the flow rate of 1.0 mL/min；the column temperature was 30 ℃；the detection wavelength was set at 242 nm；the sample size was 20 μL. Using the contents of above 4 components as evaluation indexes ，radar image analysis ，cluster thermal map analysis and principal component analysis were performed for comparing the quality of C. rotundus from different origins. RESULTS ：The results of content determination methodology investigation met relevant requirements ；the total contents of 4 components in volatile oil from C. rotundus from different origins ranged from 136.986 4 to 538.832 1 mg/g，of which the total content of samples from Yunnan was the highest （the average value was 476.059 2 mg/g）. Radar image analysis results showed that the whole contour in the 4 origins of Guangdong ， Jiangxi，Guangxi and Yunnan was large relatively and better balanced ，among which the samples from Yunnan had the largest overall contour and the best balance. The cluster thermal map analysis results showed that the samples from 12 origins could be grouped into 2 categories，the first category was from Hubei ，Jiangxi，Yunnan，Sichuan，Guangdong，Shandong，Henan and Shaanxi；the second category was from Guangxi ，Shanxi，Anhui and Hainan ；the quality of samples from the first category were better than that of samples from the second category. The principal component analysis results showed that the cumulative contribution rate of the first three principal components was 96.1％，and the samples from 12 origins were mainly clustered into two categories ，which was consistent with the results of cluster thermal map analysis. CONCLUSIONS ：Established HPLC method can be used for simultaneous determination of cypermenone ，nootkatone，α-cyperone and aristolone in volatile oil of C. rotundus from different origins. Among the samples from 12 origins，the quality of medicinal material from Yunnan is better.
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OBJECTIVE：To compare the c ontents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products，and to provide reference for the optimization of salt-processing technology. METHODS ：Processed with bittern ， magnesium chloride ，sodium chloride ，calcium chloride and potassium chloride as excipients ，and HPLC method were adopted to determine the contents of 6 kinds of ester alkaloids in raw aconite and 5 kinds of salt-processed products. The effect/toxicity ratio and toxicity component index were used to evaluate the effect and toxicity of raw aconite and 5 kinds of salt-processed products. RESULTS： The linear range of benzoylneoaconitine ， benzoylhypoconitine， benzoylaconitine， neoaconitine， aconitine and hypoconitine were 2.440-24.40，2.240-22.40，2.020-20.20，2.780-27.80，2.240-22.40，2.240-22.40 μg/mL（r≥0.999 0）. RSDs of precision，stability and repeatability tests were all less than 2％. The recovery rates were 102.13％-104.87％（RSD＝0.98％，n＝ 6），100.00％-105.00％（RSD＝2.02％，n＝6），95.00％-99.29％（RSD＝1.77％，n＝6），100.41％-104.58％（RSD＝1.78％，n＝ 6），98.87％-99.90％（RSD＝0.41％，n＝6），100.20％-104.00％（RSD＝1.55，n＝6），respectively. The contents of total alkaloids in order from the largest to the smallest was as follows ：raw aconite ＞processed products of sodium chloride ＞processed products of bittern ＞processed products of potassium chloride ＞processed products of calcium chloride ＞processed products of magnesium chloride. The order of effect/toxicity ratio was processed products of potassium chloride ＞processed products of magnesium chloride＞processed products of sodium chloride ＞processed products of calcium chloride ＞processed products of bittern ＞raw aconite. The order of toxicity component index was raw aconite ＞processed products of sodium chloride ＞processed products of bittern＞processed products of calcium chloride ＞processed products of potassium chloride ＞processed products of magnesium chloride. CONCLUSIONS ：Using 5 kinds of salt as excipients ，the proce ssing technology has different degrees of “efficacy enhancing and toxicity reducing ”effect. Among them ，magne- sium chloride ，potassium chloride and calcium chloride are better for processing and can be used for processing aconite.
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AIM: To study the chemical components from the bioactive extract of Chuanminshen violaceum Sheh et Shan in Sichuan province. METHODS: The isolation and purification of this extract were conducted by means of silica gels column chromatography.The structures of the compounds were elucidated based on their physical,chemical features and spectrum data. RESULTS: 8 compounds were isolated from the extract.They were elucidated as hexadecanoic Acid(A),2,6-dimethoxybenzoic acid(B),succinic acid (C),combination of sigmasterol and sitosterol(1(∶)1)(D),chrysophanol(E),8-hydroxy-5-methoxypsoralen(F),4-hydroxy-3-methoxybenzoic acid(G) and stigmasteryl 3-O-?-D-glucopyranoside(H). CONCLUSION: Compounds B,C,E,G were isolated from Chuanminshen violaceum for the first time.