OBJECTIVESpecies containing extremely aromatic compounds in leaves of Chimonanthus was analyzed to evaluate its genetic diversity and genetic relationships.

METHODAFLP molecular marker technique was used in the study, UPGMA cluster analysis was conducted with the software of POPGENE32.

RESULTSFive hundred and fifty-nine bands were amplified by 10 pairs of primers screened, of which 226 bands were polymorphic, and the percentage of polymorphic bands was 36.8%. Observed number of alleles, effective number of alleles, Nei's genetic diversity index and Shannon's information index were 1.992 6, 1.306 5, 0.199 2 and 0.325 1, respectively. Genetic distances of the 21 populations were ranged from 0.039 2 to 0.289 4.

CONCLUSIONSpecies containing extremely aromatic compounds in leaves of Chimonanthus with low genetic diversity play an important role in enhancing the protection of species and germplasm resources. Form the molecular level, the studies demonstrated the correctness of the result by Zhang Ruohui that species containing extremely aromatic compounds in leaves of Chimonanthus were divided into Ch. salicifolius, Ch. Zhejiangensis, Ch. nitens and Ch. grammatus.

This study was aimed to establish the HPLC fingerprints of the genus Chimonanthus leaves and compare the constituents distribution among five Chimonanthus species . The analysis was conducted on a C15 column (4.6 mm x 250 mm, 5 microm) with the mobile phase containing acetonitrile-water in gradient program: acetonitrile (B), 0-20 min, 6%-20%; 20-30 min, 20%-25%; 30-40 min, 25%-45%; 40-50 min, 45%-80%; 50-80 min, 80%-85%; 80-90 min, 85%-100%; 90-110 min, 100%. Flow rate was 0.8 mL x min(-1) and detection wavelength was 228 nm. Column temperature was set at 30 degrees C. The HPLC fingerprints of the five Chimonanthus species have been established. Ch. praecox, Ch. nitens, Ch. salicifolius, Ch. Zhejiangensis and Ch. grammatus have significant difference in constituents distribution and contents. Five standard substances as common compounds were confirmed in chromatography fingerprints. The method can be used as quality evaluation and classicfication of the genus Chimonanthus.
Calycanthaceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry

To explore anti-tumor active components of Chimonanthus salicifolius, the phytochemistry of the chloroform fraction from leaves extract was investigated by repeated silica gel column chromatography. Twelve compounds were isolated and their structures were identified by physicochemical properties and spectroscopic data analysis as 9-epi-blumenol C(1), blumenol C(2), (+)-dehydrovomifoliol (3), (+)-vomifoliol (4), robinlin (5), (-)-loliolide (6), isofraxidin (7), scopoletin (8), 6,7-dimethoxycoumarin (9), 6, 7, 8-trimethoxycoumarin (10), beta-sitostenone (11), and beta-stigmasterol(12). Compounds 1-6 belonging to nor-sesquiterpenoids were isolated from the family Calycanthaceae for the first time. Compound 1 was a new natural product. Compounds 7, 11 and 12 were obtained from this plant for the first time.
Antineoplastic Agents ; analysis ; isolation & purification ; Calycanthaceae ; chemistry ; Chloroform ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Leaves ; chemistry

The chemical composition of essential oil, which from the leaves of Chimonanthus grammatus obtained by hydrodistillation were analyzed by GC-MS, and their possible antibacterial properties were screened. According to the results from GC-MS analysis, fifty-three components comprising 99.99% of the essential oil were identified. The major components of essential oil were 3-(4, 8-dimethyl-3,7-nonadienyl)(E) -furan (13.1%), bornyl acetate (12.66%), and 6,6-dimethyl-3-methylene-bicyclo[3.1.1] heptane (7.06%), etc. Antibacterial activity of essential oil was employed by two complementary test systems of disc diffusion and MIC/ MBC tests, which showed obviously antibacterial activity against all of the tested bacteria.
Anti-Bacterial Agents ; pharmacology ; Calycanthaceae ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Oils, Volatile ; analysis ; pharmacology ; Plant Leaves ; chemistry

Sinocalycanthus chinensis, an endangered species endemic to China, is cultivated as an ornamental landscape tree in China. However, S. chinensis, Chimonanthus species and Calycanthus floridus are difficult to be distinguished in seedling market because of their similar morphological characters. In this study, ISSR (inter-simple sequence repeats) were applied to detect S. chinensis from its closely related species. A unique 748-bp band was found in all accessions of S. chinensis. SCAR (sequence characterized amplified regions) markers were created by cloning and sequencing the specific band, and designing a pair of primers to amplify the band of 748 bp. Diagnostic PCRs were performed using the primer pair with the total DNAs of S. chinensis, Chimonanthus species and C. floridus as templates, with only S. chinensis being able to be amplified. This amplification is not only rapid (results can be obtained in less than 3 h), but is also easy to perform. Hence it is a feasible method for identifying S. chinensis in seedling market.
Calycanthaceae ; genetics ; DNA, Plant ; genetics ; Genetic Markers ; genetics ; Plant Leaves ; genetics ; Random Amplified Polymorphic DNA Technique ; Species Specificity

Scopolin (SC-1), scopoletin (SC-2) and isofraxidin (IS-1) are the main active constituents in Chimonanthi Radix. However, the in vivo metabolism of SC-1, SC-2 and IS-1 have not been comprehensively clarified. In this study, the in vivo metabolic profiles of these three coumarins in the rat plasma, urine and feces were analyzed. Ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) method was applied to characterize the prototypes and metabolites of SC-1, SC-2 and IS-1 in rat feces, urine, and plasma after intravenous administration. A total of 11 metabolites of the three parent compounds were tentatively identified. The main metabolic pathways were analyzed by identification of metabolites, and it was found that these three coumarins underwent multiple in vivo metabolic reactions including glucuronidation, sulfonation, isomerism and reduction. In this study, the analysis of metabolites of three coumarins basically demonstrated their in vivo metabolic process, providing basis for the further pharmacokinetics and pharmacological evaluations of SC-1, SC-2 and IS-1.
Animals ; Calycanthaceae ; chemistry ; Chromatography, High Pressure Liquid ; Coumarins ; metabolism ; pharmacokinetics ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Rats ; Tandem Mass Spectrometry

The vapour distillation was used to extract the volatile oil of Chimonanthus salicifolius with different storage time, determine the content of cineole in volatile oil by GC, to study the influence of storage time on the content of volatile oil and cineole of C. salicifolius. We found that the content of volatile oil in fresh herbs of C. salicifolius was 0.023 0 mL x g(-1), it was decreased to 0.020 0, 0.017 5 mL x g(-1) respectively after storing for 4, 12 months; the GC methodological study of precision, stability and repeatability, RSD < 2%, the average recovery rate was 99.50%, RSD 1.7%; the content of cineole in fresh volatile oil was 54.30%, it was increased to about 62% and remained stably with the time. Therefore, the content of volatile oil and cineole of C. salicifolius can change with the storage time; GC method for the determination of the content of cineole is accurate, reliable, specific and repeatable, it's suitable as a quality control method of C. salicifolius.
Calycanthaceae ; chemistry ; Chromatography, Gas ; Cyclohexanols ; analysis ; Drug Storage ; methods ; Drugs, Chinese Herbal ; analysis ; Monoterpenes ; analysis ; Oils, Volatile ; analysis ; Plant Oils ; analysis

Glucose-6-phosphate dehydrogenase is main regulatory enzyme for pentose phosphate pathway. To amplify the core sequence of G6PDH gene from Chimonanthus praecox, the primers were synthesized, based on the conserved nucleotide sequence of other reported plant G6PDH genes. The specific primers were designed according to the major fragment. The full length cDNA of the G6PDH1 gene was isolated by the 3' and 5' rapid amplification of cDNA ends approach. Transcript levels of G6PDH1 isoform was measured by real-time quantitative RT-PCR in different tissues and in responds to cold treatment. The G6PDH1 subcellular localization, transmembrane domain, three-dimensional structure, and phylogenetic analysis were predicted by different software to analysis the bioinformatics of G6PDH1 protein. The G6PDH1 cDNA sequence was 2 011 bp in length and consisted of 1 551 bp Open Reading Frame (ORF) , encoding a protein of 516 amino acids. Expression analysis results in different tissues showed that G6PDH1 was primarily observed in flowers and roots, as opposed to the leaves and stems. Cold treatment experiments indicated that cold treatment caused a rapid increase in G6PDH1 expression in flowers within 12 h. The full-length cDNA of G6PDH1 and its expression analysis will play an important role for further study on cold stress responses in Ch. praecox.
Calycanthaceae ; chemistry ; classification ; enzymology ; genetics ; Cloning, Molecular ; Enzyme Stability ; Glucosephosphate Dehydrogenase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism

Chimonanthus plants widely distributed in southern area of China, which have a long history of edibles and medicine. Phytochemical investigations have shown that Chimonanthus produced 143 non-volatile constituents, including alkaloids, flavonoids, terpenoids, coumarins and others, which exhibit significant anti-oxidant, anti-bacterial, anti-cancer, anti-inflammatory, antihyperglycemic, antihyperlipidemic and other biological activities. On the basis of systematic reviewing of literatures, this article overviews the non-volatile constituents and pharmacology of Chimonanthus from domestic and foreign over the last 30 years (until June 2018), and may provide a useful reference for the further development of Chimonanthus.
Animals ; Calycanthaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; toxicity ; Humans ; Medicine, Chinese Traditional ; Phytochemicals ; chemistry ; pharmacology ; therapeutic use ; toxicity ; Phytotherapy

In the present study, four new sesquiterpenoids, chimonols A-D (compounds 1-4), together with four known compounds (5-8) were isolated from the EtOAc extract of Chimonanthus praecox Link. The structures of these new compounds were elucidated on the basis of spectroscopic techniques (UV, IR, MS, and 1D and 2D NMR), and their absolute configurations were established by comparing experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 1-8 were evaluated for antimicrobial activities and the minimum inhibitory concentrations (MICs) were determined by the broth microdilution method in 96-well culture plates. Compounds 1, 2, and 7 exhibited weak antibacterial effects for S. aureus (ATCC 6538), E. coli (ATCC 11775), and P. aeruginosa (ATCC 10145) with MIC values being 158-249 µg·mL. Compounds 3-7 showed activities against C. glabrata (ATCC 2001) and S. aureus (ATCC 43300) with MIC values being 128-197 µg·mL. Compounds 1-4 showed activity against S. aureus (ATCC 25923) with MIC values being 162-254 µg·mL. The present study provided a basis for future evaluation of these compounds as antibacterial agents.
Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Calycanthaceae ; chemistry ; Escherichia coli ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Sesquiterpenes ; chemistry ; isolation & purification ; pharmacology ; Staphylococcus aureus ; drug effects