ABSTRACT
Objective: The experiment was designed to reveal the extraction, distribution and influencing factors of volatile components in the extraction process of volatile oil from Acori Tatarinowii Rhizoma (ATR). Methods: Volatile oil of ATR was extracted by steam distillation and the extract was collected every 30 min to separate the aromatic water and volatile oil. Results: A total of 56 volatile compounds were determined, of which β-asarone, methyleugenol, cis-methylisoeugenol and γ-asarone were the main characteristic constituents. There were 41 kinds of components distributed only in water, four components only in oil and 11 kinds in both oil and water. Correlation analysis showed that the specific components in water were positively correlated with the dissolution/diffusion of the main components in water, but negatively correlated with the main components in volatile oil. The water solubility of the unique components in water was the highest. The results of radar and PCA showed that the water solubility and boiling point of the specific components in water were very high, the vapor pressure of the common components of oil and water was the highest, and the polar surface areas of the special components in oil were high. Conclusion: Affected by the physical and chemical properties of volatile component, some components specifically distributed in water increased the content of main components in the aromatic water, may resulting in volatile oil extraction process easy to "emulsification", in turn, leading to an important reason for the declining quality of volatile oil.
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Objective: To clone the enzyme genes related with methyl eugenol synthesis and characterize the corresponding sequence information based on the transcriptome sequencing of Asarum heterotropoides. Methods: RT-PCR was performed to obtain the full length cDNA of the phenylalanine lyase (PAL) gene, cinnamic acid-4-hydroxylase (C4H) gene, 4-hydroxycinnamoyl-CoA ligase (4CL), and cinnamyl alcohol dehydrogenase (CAD) genes by using young leaves as materials. The acquired enzyme genes were analyzed by bioinformatics. Results: The ORF lengths of AhPAL, AhC4H, Ah4CL, and AhCAD were 2 157, 1 278, 1 623 and 1 071 bp, which respectively encoded 718, 425, 540, and 356 amino acids. Four proteins had respective conserved domains. The amino acid sequences of AhPAL, Ah4CL, and AhCAD were similar to those of other reported species except for AhC4H. Conclusion: Four enzyme genes related with methyl-eugenol synthesis in A. heterotropoides were separated and analyzed using bioinformatics method. These results would lay the important foundation for functional analysis of corresponding genes and for elucidating the regulation mechanism of methyl-eugenol biosynthesis.
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Objective: To obtain the transcriptome sequence database and differentially expressed genes of Asarum sieboldii, and to identify the genes related to the biosynthesis of methyleugenol, the main chemical compound in this species. Methods: Roots and leaves of the plants were chosen as experiment materials. The transcriptome sequence database was constructed by applying an Illumina Hiseq 4000 Sequencing Platform. Unigenes were assembled by BLAST similarity searches and annotated with GO and KEGG orthologs identifiers. Moreover, differentially expressed genes were analyzed. Results: 12.25 Gb database was obtained, among which 129 003 unigenes were annotated to be involved in 52 GO-terms and 363 metabolic pathways. After analysis, 439 differentially expressed genes were observed, the up-regulated genes account for 38.3% and the down-regulated genes account for 61.7%. In addition, 136 unigenes involved in phenylpropanoid biosynthesis in A. sieboldii, and 44 unigenes that were associated with biosynthesis of methyleugenol were identified. Conclusion: Unigenes explored in this study will significantly contribute to genome-wide research and analysis of this species.
ABSTRACT
Croton malambo is a plant used in traditional medicine, in Colombia. The aim of this research was to characterize the essential oils (EO) from leaves and branches by GC-MS, NMR and determine the antiradical capacities and the in vitro and in vivo cytotoxic properties of the EO, methyleugenol (ME) and eugenol (EU). The EO of leaves and branches of C. malambo presented to ME as the main constituent (68.5 percent and 85.1 percent, respectively) and their structure was confirmed by NMR. On the other hands, the antiradical capacities (ABTS+. method) of the EO and ME were very low, obtaining only inhibition values at a fixed concentration: to 2045 ug/mL 50 +/- 2 percent (leaves EO) and 28 +/- 1 percent (branches EO); and, 2218 ug/mL - 2.0 +/- 0.2 percent (ME). While EU had the highest value of TAA (14003 +/- 719 mmol Trolox®/kg SE). According to lymphocytes citotoxicity test, all tested substances were classified as moderately toxic, with values of LC50 between 310 +/- 17 897 +/- 11 ug/mL, being the EO the most toxic. The assessment of the toxicity in Zebra fish embryos indicated that LC50 of the branches EO, ME and EU were between 16 +/- 9 43 +/- 9 ug/mL, being the EU the most toxic.
Croton malambo es una planta empleada en medicina tradicional, en Colombia. El objetivo de esta investigación fue caracterizar los aceites esenciales (AE) de hojas y ramas por GC-MS, RMN y determinar las capacidades antiradicalarias y las propiedades citotóxicas in vitro e in vivo de los AE, metileugenol (ME) y eugenol (EU). Los AE de hojas y ramas de Croton malambo presentaron a ME como el constituyente principal (68.5 por ciento y 85.1 por ciento, respectivamente) y su estructura fue confirmada por RMN. Por otro lado, las capacidades antiradicalarias (método ABTS+.) de los AE y ME fueron muy bajas, obteniéndose sólo valores de inhibición a una concentración fija: a 2045 ug/mL 50 +/- 2 por ciento (AE de hojas) y 28 +/- 1 por ciento (AE de ramas); y, 2218 ug/mL - 2.0 +/- 0.2 por ciento (ME). Mientras que, EU tuvo el mayor valor de TAA (14003 +/- 719 mmol Trolox®/kg SE). Según el ensayo de citotoxicidad en linfocitos, todas las sustancias evaluadas se catalogaron como moderadamente tóxicas, con valores de CL50 entre 310 +/- 17 897 +/- 11 ug/mL, siendo los AE los más tóxicos. La estimación de la toxicidad en embriones del pez Cebra indicó que las CL50 del AE ramas, ME y EU estuvieron entre 16 +/- 9 43 +/- 9 ug/mL, siendo el EU el más tóxico.
Subject(s)
Oils, Volatile/pharmacology , Croton/chemistry , Eugenol/pharmacology , Plant Leaves/chemistry , Antioxidants , Oils, Volatile/chemistry , Gas Chromatography-Mass SpectrometryABSTRACT
The chemical composition of the essential oil from the leaves of Pelargonium odoratissimum (L.) L'Hér., Geraniaceae, was determined and the antimicrobial activities against the Aspergillus flavus CML 1816, Aspergillus carbonarius CML1815 and Aspergillus parasiticus CMLA 817 fungi, as well the Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25 992 bacteria were evaluated. The essential oil was isolated by steam distillation using a modified Clevenger apparatus, and its constituents were identified and quantified by GC/MS and GC-FID analyses. In vitro bioanalytical testing was performed using a completely randomized design. The concentrations of essential oil employed ranged from 0.1 to 2 μL.mL-1 (in dimethyl sulfoxide) for the fungus species and from 1 to 500 μL.mL-1 for the bacteria. The diameters of the inhibition zones formed for bacteria and the mean diameters of mycelial growth in perpendicular directions for fungi were measured, followed by calculation of the percentage of inhibition. The essential oil from the leaves of P. odoratissimum furnished methyleugenol (96.80 percent), a phenylpropanoid. This essential oil inhibited the growth of fungi (100 percent inhibition) and exhibited a small effect on the bacteria at the concentrations tested.