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1.
J Chromatogr A ; 1612: 460630, 2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-31677768

RESUMO

Authentication of original species is embedded in the quality control system of herbal medicines. In this work, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based untargeted metabolomics coupled with chemometric analysis was utilized for the precise authentication of the Fritillaria species for both raw materials and commercial products. First, a stepwise difference-enlarging chemometric analysis strategy was proposed to analyze eight medicinal Fritillaria species. Subsequently, 21 species-specific markers were discovered and the specificity was investigated under different sample preparation methods. Finally, the obtained species-specific markers were successfully utilized to identify the Fritillaria species in commercially relevant products. This work is the first to report robust and specific markers for authentication of Fritillaria products, showing promise for tracking the supply chain of herbal suppliers.


Assuntos
Alcaloides/análise , Fritillaria/química , Esteroides/análise , Cromatografia Líquida , Fritillaria/metabolismo , Espectrometria de Massas , Metabolômica , Plantas Medicinais/química , Plantas Medicinais/metabolismo , Especificidade da Espécie
2.
Genomics ; 111(6): 1360-1372, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-30248489

RESUMO

Crown imperial (CI) has been used in traditional medicine. Today it is known that such beneficial effects are due to its richness in steroidal alkaloids (SA). Using de novo transcriptomics, orthologues/paralogues finder, phylogenetic analysis and tissue- and developmental stage-specific expression analysis, we identified ten genes and several TFs involved in the biosynthesis of SA in CI. The comparative analysis of ten genes expression profiles revealed the possibility of their co-regulation, which may imply the possibility of their organization in metabolic gene clusters. Having in mind convergent evolution of steroidal biosynthetic pathways in flowering plants and records of convergent evolution of specific proteins, observed expression patterns open a reasonable interest to investigate the possibility of the existence of genes cluster organization in SA pathway in the family Liliaceae or at least in some species of genus Fritillaria. Obtained results support transcriptomics as useful approach in elucidating genes underlying complex biochemical pathways.


Assuntos
Alcaloides/biossíntese , Fritillaria/genética , Fritillaria/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Esteroides/biossíntese , Transcriptoma , Alcaloides/genética , Vias Biossintéticas , Biologia Computacional , Fritillaria/crescimento & desenvolvimento , Filogenia , Proteínas de Plantas/metabolismo , Esteroides/metabolismo
3.
J Food Drug Anal ; 26(1): 337-347, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29389572

RESUMO

Thunberg fritillary bulb (the dry bulbs of Fritillaria thunbergii Miq.), a traditional Chinese Medicine, is widely applied as an expectorant and antitussive. In this investigation, the primary metabolites of bulbs, flowers, leaves, and stems of F. thunbergii were analyzed by gas chromatography-mass spectrometry. Principal component analysis, partial least squares-discriminate analysis, orthogonal projection to latent structures-discriminate analysis, and heat map analysis showed that there were dissimilar metabolites, and a negative correlation between amino acids and saccharides in different analytes. Furthermore, carbodiimide, tryptophan, glucose-6-phosphate, xylose, 2-piperidinecarboxylic acid, monoamidomalonic acid, phenylalanine, and histidine were found to play an important role in the plant metabolism net of F. thunbergii.


Assuntos
Fritillaria/química , Cromatografia Gasosa-Espectrometria de Massas , Metaboloma , Metabolômica , Biologia Computacional/métodos , Análise de Dados , Fritillaria/metabolismo , Redes e Vias Metabólicas , Metabolômica/métodos
4.
Phytochem Anal ; 29(3): 290-299, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29336082

RESUMO

INTRODUCTION: Bulbus fritillariae cirrhosae (known as Chuan bèi mǔ in China, BFC) contain fritillaria steroidal alkaloids as the bioactive ingredients and are widely used as traditional Chinese medicine for the treatment of cough and phlegm. Due to limited wild resources, the cultivated species are becoming predominantly used in Chinese traditional medicine markets. OBJECTIVE: To assess the impact of different growth stages on the alkaloids of cultivated BFC and establish a reference for quality control and guidance for appropriate harvesting practices. METHODS: The ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) metabolomic strategy was applied to determine potential chemical markers for the discrimination and quality control of cultivated BFC in different growth stages. The molecular feature extraction and multivariate statistical analysis were applied to alkaloid extraction and full metabolomic profiling of cultivated BFC for classification and marker compound characterisation. RESULT: This approach allowed the establishment of a fast and efficient comparative multivariate analysis of the metabolite composition of 42 samples covering growth of cultivated BFC ranging in age from one to seven years old. Four alkaloid compounds were identified in cultivated BFC based on accurate mass, retention time, and MS/MS fragments. These compounds may be used as potential chemical markers for the classification and discrimination of cultivated BFC samples indifferent growth stages. CONCLUSIONS: The proposed analytical method in combination with multivariate statistical analysis comprised a useful and powerful strategy to explore the chemical ingredients and transforming mechanisms of cultivated BFC and for quality evaluation and control. Copyright © 2018 John Wiley & Sons, Ltd.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Fritillaria/metabolismo , Metabolômica , Espectrometria de Massas em Tandem/métodos , Alcaloides/isolamento & purificação , Fritillaria/crescimento & desenvolvimento , Medicina Tradicional Chinesa , Análise Multivariada , Controle de Qualidade , Padrões de Referência
5.
Zhong Yao Cai ; 34(2): 183-6, 2011 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-21823471

RESUMO

OBJECTIVE: Set up Fritillaria cirrhosa cell mass suspension culture system to rapidly screen the best culture conditions for cell mass proliferation and hormone combination. METHODS: Using MS medium as the basic medium, the impact of inoculum size, hormone combination, growth regulators for Fritillaria cirrhosa cell mass suspension culture were compared, and also the growth of cell mass at different culture conditions was compared, and the total alkaloids content in proliferative cell mass was measured. RESULTS AND CONCLUSION: Fritillaria cirrhosa grow significantly faster in cell mass suspension culture than in the solid culture. The total alkaloid content in cell mass is higher than commercial and wild bulb of Fritillaria cirrhosa. The optimal inoculum size for cell mass suspension culture is 30 g/L and the optimal culture media is MS +6-BA 2.0 mg/L + NAA 0.2 mg/L.


Assuntos
Alcaloides/metabolismo , Técnicas de Cultura de Células/métodos , Fritillaria/citologia , Fritillaria/metabolismo , Reguladores de Crescimento de Planta/farmacologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas/metabolismo , Meios de Cultura , Fritillaria/crescimento & desenvolvimento , Reguladores de Crescimento de Planta/metabolismo , Ribavirina/metabolismo , Ribavirina/farmacologia , Espectrofotometria
6.
Oecologia ; 165(4): 847-54, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21190042

RESUMO

Evolution of dispersal is affected by context-specific costs and benefits. One example is sex-biased dispersal in mammals and birds. While many such patterns have been described, the underlying mechanisms are poorly understood. Here, we study genetic and phenotypic traits that affect butterfly flight capacity and examine how these traits are related to dispersal in male and female Glanville fritillary butterflies (Melitaea cinxia). We performed two mark-recapture experiments to examine the associations of individuals' peak flight metabolic rate (MR(peak)) and Pgi genotype with their dispersal in the field. In a third experiment, we studied tethered flight in the laboratory. MR(peak) was negatively correlated with dispersal distance in males but the trend was positive in females, and the interaction between MR(peak) and sex was significant for long-distance dispersal. A similar but nonsignificant trend was found in relation to molecular variation at Pgi, which encodes a glycolytic enzyme: the genotype associated with high MR(peak) tended to be less dispersive in males but more dispersive in females. The same pattern was repeated in the tethered flight experiment: the relationship between MR(peak) and flight duration was positive in females but negative in males. These results suggest that females with high flight capacity are superior in among-population dispersal, which facilitates the spatial spreading of their reproductive effort. In contrast, males with high flight capacity may express territorial behaviour, and thereby increase the number of matings, whereas inferior males may be forced to disperse. Thus, flight capacity has opposite associations with dispersal rate in the two sexes.


Assuntos
Borboletas/metabolismo , Fritillaria/metabolismo , Migração Animal , Animais , Borboletas/genética , Borboletas/crescimento & desenvolvimento , Feminino , Fritillaria/crescimento & desenvolvimento , Genótipo , Glucose-6-Fosfato Isomerase/genética , Glucose-6-Fosfato Isomerase/metabolismo , Masculino , Reprodução , Comportamento Sexual Animal
7.
Zhongguo Zhong Yao Za Zhi ; 36(18): 2462-4, 2011 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-22256745

RESUMO

OBJECTIVE: To establish cryopreservation system of shoot-tips from Fritillaria anhuiensis. METHOD: Taking vitrification as system of cryopreservation, the shoot tips with length 2-3 mm were precultured in MS medium enriched with 0.4 mol x L(-1) sucrose for 3 d. They were treated for 20 min with 60% PVS2 at 25 degrees C, and then subjected to ice-cooled vitrification solution for 60 min and transferred to 2 mL cryotubes with fresh vitrification solution (PVS2) and plunged into liquid nitrogen. After rapid thawing in 40 degrees C water bath for 1 min, shoot-tips were expelled into MS medium containing 1.2 mol x L(-1) sucrose for 20 min. Further recovery and growth took place on regeneration medium in the dark at 25 degrees C for 2 weeks, and then with light/dark cycle of 12/12 h. The genetic integrity of cryopreserved shoot-tips was identified through products of PCR with arbitrary primers. RESULT AND CONCLUSION: The highest survival rates of shoot-tips reached 79.9% by vitrification, and the regeneration rates were 52.3%. No changes were found between treated materials and untreated materials in genomic DNA.


Assuntos
Criopreservação/métodos , Fritillaria/metabolismo , Brotos de Planta/metabolismo , Plantas Medicinais/metabolismo , Vitrificação , Crioprotetores/química , Fritillaria/genética , Instabilidade Genômica/genética , Brotos de Planta/genética , Plantas Medicinais/genética , Preservação Biológica , Análise de Sobrevida
8.
Zhong Yao Cai ; 33(6): 854-6, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-21049604

RESUMO

OBJECTIVE: The optimal in vitro culture conditions for the regeneration of Fritillaria cirrhosa were screened. METHODS: Through the in vitro culture of the flowering stage Fritillaria cirrhosa, the effect of illumination, culture temperature, hormone combination and the amount of ribavirin on the regeneration of Fritillaria cirrhosa were studied. RESULTS AND CONCLUSION: The explant browning level could be apparently reduced if the bulb were cultured under the ray after exposure to proper low-temperature. The optimal temperature for the regeneration of bulb is (20 +/- 2) degrees C. The hormone has obvious promotional effect on the regeneration of bulb, and the best combination is 6-BA 2.0 mg/L plus NAA 0.2 mg/L Concentration of 10 mg/L of ribavirin is best for the regeneration of bulb.


Assuntos
Fritillaria/crescimento & desenvolvimento , Fritillaria/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Ribavirina/metabolismo , Conservação dos Recursos Naturais , Luz , Reguladores de Crescimento de Planta/química , Raízes de Plantas/metabolismo , Regeneração , Ribavirina/química , Temperatura , Técnicas de Cultura de Tecidos/métodos
9.
J Sep Sci ; 33(11): 1587-94, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20405484

RESUMO

A sensitive and reliable HPLC-diode-array detector method was developed for the first time to simultaneously determine nine nucleosides and nucleobases including uracil, cytidine, guanine, uridine, thymine, inosine, guanosine, thymidine and adenosine in 13 different Fritillaria species. The analysis was performed on a BaseLine C18 column with a gradient of acetonitrile in water at a flow rate of 0.8 mL/min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. Satisfactory separation of these compounds was obtained in less than 40 min. The optimized method provided good linear relation (r(2)>0.9995 for all the investigated analytes), satisfactory precision (RSD <1.51%) and good recovery (from 97.64 to 101.16%). The established method was successfully applied to simultaneous determination of nine nucleosides and nucleobases in 61 batches of samples from 13 Fritillaria species collected from different habitats in China, which could be helpful to control the quality of Fritillaria bulbs.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Fritillaria/metabolismo , Nucleosídeos/química , Calibragem , Química , Técnicas de Química Analítica , China , Modelos Químicos , Nucleotídeos/química , Extratos Vegetais/química , Reprodutibilidade dos Testes , Solventes/química , Fatores de Tempo
10.
Zhongguo Zhong Yao Za Zhi ; 33(16): 1948-51, 2008 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-19086625

RESUMO

OBJECTIVE: To study the physiologic indexes, yield and the contents of alkaloids of Fritillaria cirrhosa D. Don under different temperature. METHOD: The growth temperatures (15, 20, 25, 30 degrees C) of F. cirrhosa were controlled by using artificial climate, the growth was observed, the contents of chlorophyll a and b, soluble sugar, MAD, proline of the leaves of F. cirrhosa were tested, and the yield and the alkaloids content of the bulbs were analyzed. RESULT: The growth period of F. cirrhosa under 15, 20 degrees C were appropriately extended. The difference of the content of leaves chlorophyll b under four temperatures and the contents of total chlorophyll and chlorophyll a under 15, 20, 30 degrees C were not significant. The contents of soluble sugar, MAD and proline of leaves and the growth ratio, dry weight and content of alkaloids of bulb increased with the temperature decrease. CONCLUSION: Higher temperature is not suitable for the growth of F. cirrhosa. Under the relatively lower temperature, the growth period of F. cirrhosa extended, the bulb can grow properly, and the content of alkaloid increased. F. cirrhosa can improve its cold tolerance by increasing the content of proline and soluble sugar, and it also can maintain the normal content of chlorophyll under the lower temperature.


Assuntos
Alcaloides/metabolismo , Fritillaria/crescimento & desenvolvimento , Fritillaria/metabolismo , Regulação da Expressão Gênica de Plantas , Temperatura
11.
J Huazhong Univ Sci Technolog Med Sci ; 28(1): 118-20, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18278474

RESUMO

A reverse-phase high pressure liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) has been developed for the quantitative analysis of hupehenine in the total alkaloids from Fritillaria hupehensis. Samples were analyzed on a reverse-phase column (Hypersil C-18) with a mobile phase of methanol:water:chloroform: triethylamine (85:15:1:0.6). The ELSD was set at the drift tube temperature of 68.3 degrees C and gas flow rate of 1.8 L/min. Hupehenine's retention time was 13.7 min with an asymmetry factor of 1.2. The validity of the method has been verified with linearity, limit of detection, accuracy and precision. The logarithmic linear curve was obtained from 8.936 to 134.04 microg/mL (r=0.9993). The detection limit (S/N>3) of hupehenine was 1.79 microg/mL on the column. Intra-day RSD was 1.42% and inter-day RSD was 2.26% (3 days within a week). The average recovery of hupehenine was 101.50%, and RSD was 1.62%.


Assuntos
Alcaloides/química , Fritillaria/metabolismo , Extratos Vegetais/análise , Alcaloides/análise , Alcaloides/farmacologia , Cromatografia/métodos , Cromatografia Líquida de Alta Pressão/métodos , Luz , Modelos Químicos , Modelos Estatísticos , Extratos Vegetais/farmacologia , Reprodutibilidade dos Testes , Espalhamento de Radiação , Espectrofotometria Ultravioleta/métodos , Temperatura , Fatores de Tempo
12.
Z Naturforsch C J Biosci ; 63(11-12): 789-93, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-19227824

RESUMO

Ten strains of endophytic fungi were isolated from the bulbs of the traditional Chinese medicinal plant Fritillaria ussuriensis. The extract from one of them, Fu7, showed a positive reaction with Dragendorff's reagent and the same Rf value in thin-layer chromatography (TLC) analysis as authentic sipeimine. A further TLC scan and high-performance liquid chromatography-evaporative light-scattering detection (HPLC-ELSD) showed that one ingredient of the extract of strain Fu7 had a similar absorption curve in the range 200-700 nm and the same retention time as authentic sipeimine. Thus, the fungus produces the bioactive ingredient sipeimine, as does its host plant, and could be used for the production of sipeimine by fermentation.


Assuntos
Cevanas/isolamento & purificação , Cevanas/metabolismo , Fritillaria/metabolismo , China , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Delgada , Medicamentos de Ervas Chinesas/isolamento & purificação , Filtração , Fritillaria/química , Fritillaria/crescimento & desenvolvimento , Raízes de Plantas/química
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