Flavanoids from Clematis hexapetala

Aim To study the chemical constituents from the roots and rhizomes of Clematis hexapetala Pall.. Methods The components were isolated by means of solvent extraction, repeated chromatography with silica gel, sephadex LH-20 and prep-HPLC. The structures were determined by spectrum analysis. Results Twelve flavonoids were isolated and their structures were identified as 3, 5, 6, 7, 8, 3＇, 4＇-heptamethoxyflavone （1）, nobiletin （2）, liquiritigenin （3）, hesperetin （4）, naringenin （5）, liquiritigen-7-O-β-D-glucopyranoside （6）, 5,7, 4＇ -trihydroxy-3＇- methoxyflavanone -7-O-α-L-rhamnopyranosyl （ 1→6 ） -β-D-glucopyranoside （7）, 6-hydroxybiochain A （ 8）, formononetin （9）, daidzein （10）, genistein （11）, and tectoridin （12）. Conclusion All the said compounds were isolated from the plant for the first time.

Leaf Epidermal Feature in Clematis (Ranunculaceae) with Reference to Its Systematic Significance

For finding valuable systematic characters, leaf epidermis of 77 taxa, representing 12 sections of the genus Clematis and three related genera in the Ranunculaceae, were examined mainly by means of light microscopy (LM), and partially by scanning electron microscopy (SEM). It was shown that the leaf epidermal cells were usually irregular or polygonal in shape. The patterns of anticlinal walls were straight, arched or undulate. The stomatal apparatus is anomocytic and exists in abaxial epidermis of all species, and in the adaxial epidermis of some species. Under SEM observation, the leaf epidermis was often striated, seldom nearly smooth, and often with flakes attached. Evidence from leaf epidermis serves as a criterion for distinguishing the subsections in sect. Meclatis (Spach) Tamura and in sect. Fruticella Tamura. The results also support that there are several separate evolutionary processes in the genus Clematis .

Comparison on Four Extraction Methods of Genomic DNA from Clematis fasciculiflora Franch

[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch.[Method] Leavies of Clematis fasciculiflora Franch were used as materials for comparing the purity and concentration of extracted DNA and extracting time among the four extraction methods of genomic DNA including improved CTAB method Ⅰ,improved CTAB method Ⅱ,improved CTAB method Ⅲ and improved SDS method.[Result] The four extraction methods could all be successfully used for extracting the genomic DNA from Clematis fasciculiflora Franch.The purity of genomic DNA was the highest using improved CTAB method Ⅰ,with the longest extracting time;while the concentration of genomic DNA was the maximum using the improved SDS method,with the shortest extracting time and relatively low purity;the extracting time of improved CTAB method Ⅲ was the shortest.[Conclusion] This study had established the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch and supported for the further research using molecular biological methods.

Studies on the Chemical Components of Clematis chinensis

Two novel compounds, clemaphenol A and dihydro-4-hydroxy-5-hyroxymethyl-2(3H)-fura-none were isolated together with eight known compounds, isoferulic acid, b-sitosterol, daucosterol, 5-hy-droxymethyl-2-furancarboxaldehyde, 5-hydroxy-4-oxo-pentanoic acid, palmitic acid, linoleic acid and anemonin from the root of Clematis chinensis Osbeck.

Two Novel Antifungal Saponins from Tibetan Herbal Medicine Clematis tangutica

Antifungal assay-guided isolation of the ethanol extract of the aerial parts of Clematis tangutica yielded two novel triterpene saponins. Their structures were determined to be 3-O-a-L-arabinopyranosyl hederagenin 28-O-a-L-rhamnopyranosyl ester (1) and 3-O-b-D- glucopyranosyl(14)-a-L-arabinopyranosyl hederagenin 28-O-a-L-rhamnopyranosyl ester (2) on the basis of spectral data and chemical reactions.

A revision of Clematis sect.Atragene(Ranunculaceae)

Clematis sect. Atragene is revised in this paper based on the examination of a large number of herbarium specimens, extensive field observations, and morphometric analyses. Brief taxonomic history and geographical distribution of the section are given, the relationships among the species are discussed, and the evolutionary trends of some characters in the section are evaluated. The staminodes of the plants in this section may have evolved from the outer stamens with petaloid filaments and gradually disappearing anthers. Subsequently, they may have evolved in two different ways. One possibility is that the staminodes elongate and become lanceolate, as long as sepals, and their apices turn into attenuate. The other is that the staminodes are spathulate, but not elongating, as long as stamens, and their apices turn into retuse from obtuse and rounded. The evolutionary trend of sepals may be from thin to thick in texture, and the veins from non-prominent to prominent. As a result, five new series are established and nine species, two subspecies and nine varieties （including three new ranks） are recognized in this section. An identification key is provided, and each taxon is described and illustrated. Clematis sibirica and Clematis ochotensis are treated as subspecies of Clematis alpina due to their subtle differences and lack of, or few, overlapping distributions. Clematis fusijamana and Clematisfauriei are recognized as varieties of C. alpina ssp. ochotensis for the continuous variation of the velutinous strips on the sepal margins. Clematis iliensis is treated as variety of C. alpina ssp. sibirica for the continuous variation of leaf division types. Extensive variations in sepal color and basal caruncle size support degrading Clematis chiisanensis as a variety of Clematis koreana. The North American ser. Occientales may be primitive, whereas ser. Macropetalae may be the most advanced taxon in this section. Ser. Alpinae and ser. Koreanae are closely related to each other. However, the systematic position of ser. Tomentosae cannot be determined based on morphological characters alone in the present study.

A new triterpenoid saponin from Clematis ganpiniana

A new triterpenoid saponin, named clematiganoside A (1), was isolated from the whole plant of Clematis ganpiniana. Its structure was elucidated on the basis of 1D, 2D NMR, TOF-MS and ESI-MS techniques, and physicochemical properties.

CLEMOCHINENOSIDE A, A MACROCYCLIC COMPOUND FROM CLEMATIS CHINENSIS

The structure of a new macrocyclic compound, clemochinenoside A obtained from Clematis chinensis was identified by NMR, FAB-MS, NOE and chemical hydrolysis.

Antifungal,molluscicidal and larvicidal assessment of anemonin and Clematis flammula L.extracts against mollusc Galba truncatula,intermediate host of Fasciola hepatica in Tunisia

Objective: To investigate the potential of anemonin and Clematis flammula(C.flammula) extracts against infective organisms.Methods: The molluscicidal activities of anemonin and C.flammula extracts against Galba truncatula Mull.(Lymnaeidae) and Fasciola hepatica larval stages contaminating this snail in Tunisia were assessed by testing six groups of snails in 250 mL of extracts and aqueous dechlorinated solutions with different concentrations(ranging from 2.5 to 20.0 mg/L) for 48 h.Besides, the antifungal potential of C.flammula leaves and flowers was evaluated by using the diffusion agar and broth dilution methods against four fungal strains: Aspergillus niger, Pythium catenulatum, Rhizoctonia solani and Fusarium phyllophilum.Results: As a result, hexane and ethyl acetate flower extracts exhibited significant molluscicidal activities with LC_(50) median lethal concentrations values of 11.87 and 11.65 mg/L, respectively while LC_(50) value of anemonin was 9.64 mg/L after 48 h exposure.The flower extracts showed a larvicidal effect with a deterioration rate exceeding 35.39% where flower ethyl acetate residue gave a deterioration rate of cercariae close to 97%.Moreover, C.flammula extracts were not noxious to the associated fauna survival.All extracts inhibited the growth of P.catenulatum, the leaves and flowers methanolic extracts had the more important fungicide action with minimum inhibitory concentrations of 1.56 and 3.12 mg/mL together with minimum fungistatic concentrations of 3.12 and 6.25 mg/mL respectively.Only flower extracts were active against Rhizoctonia solani with minimum inhibitory concentrations varying between 0.70 and 1.56 mg/mL and 6.25 mg/mL of minimum fungistatic concentration.Phytochemical tests showed that the antifungal activity may be attributed to the presence of the flavonoids/saponins in the methanolic extracts and the molluscicide effects could be due to the richness of hexane and ethyl acetate extracts on sterols and triterpenoids.Conclusions: This study emphasizes the important molluscicidal and antiparasitic effects of flower ethyl acetate extracts and anemonin compound as well as the considerable antifungal activities of methanolic extracts.These results improve the therapeutic virtues of C.flammula aerial part extracts.

Triterpenoid Saponins from Clematis tangutica

Two new triterpenoid saponins, tanguticoside A and B were isolated from aerial part of Clematis tangutica. Their structures were elucidated as 3-O-beta-D-glucopyranosyl hederagenin 28 O-alpha-L-rhamnopyranosyl- (1-->4)-beta-D-glucopyranosyl- (1-->6)-beta-D-glucopyranosidel and 3-O-beta-D-glucopyranosyl ranosyl- (1-->2)-beta-D-glucopyranosyl hederagenin 28-O-alpha-L-rhamnopyranosyl- (1-->4)-beta-D-glucopyranosyl- (1-->6)-beta-D-glucopyranoside2, respectively.

NEW MACROCYCLIC COMPOUND,CLEMOCHINENOSIDE B FROM CLEMATIS CHINENSIS

The structure of a new macrocyclic compound,clemochinenoside B, isolated from the root of Clematis chinensis was elucidated by means of NMR,FAB-MS,NOE and chemical hydrolysis.

Tetraploid Induction and Cytology Identification of Clematis heracleifolia

[ Objective] This study aimed to induce the tetraploid of Clematis heracleifolia. [ Method ] Mixture of 0.2% colchicine solution and 1% agar were heated into a gel smeared on the apical meristem top bud of Clematis heracleifolia seedling to induce its tetraploid. The induction lasted for 24, 48 and 72 h respectively in three treatments to determine the best induction time. Finally, the morphological and cytological characteristics were identified and compared between the mutants and the controls. [Result] The variation rate was the highest, up to 80% when the induction time was 48 h. Compared with controls, the leaf of mutant was longer, wider and thicker, and the leaf index was smaller. The stomata size and density of lower leaf epidermis between the mutants and controls were significantly different. Chloroplast number in guard cells and chlorophyll content of mutants were all increased significantly compared with the controls through microscope observation of lower leaf epidermis and SPAD value. Chromosome number of most mutants was 32 （2n =4x）, while that of controls was 16 （2n =2x） by cytology observation of root tip cells. [ Conclusion] Tetraploid of Clematis heracleifolia was successfully obtained.

铁线莲属植物花部表型性状多样性分析及综合评价

从123份铁线莲样本中选择13个花部表型性状进行多样性分析、相关性分析、主成分分析、聚类分析、逐步回归分析和综合排名,结果表明,各性状间存在较大变异,其中6个数量性状的变异系数为24.79%~181.18%,平均变异系数为63.84%,单花萼瓣数变异系数最大(181.18%),萼瓣纵径变异系数最小(24.79%)。各性状间遗传多样性指数为5.688~6.915,以花色中线的遗传多样性指数最高(6.915),单花萼瓣数的遗传多样性指数最低(5.69)。各性状间也存在程度不等的相关性,以萼片类型与单花萼瓣数、花径与萼瓣纵径、花径与萼瓣横径、萼瓣纵径与萼瓣横径、花径与花色中线这几组间的相关性最为显著。基于主成分分析并结合聚类分析,排序得到各大类群中性状突出的种质资源,如针瓣皇帝Clematis Needle Kaiser、宽瓣皇帝C.Broad Kaiser、水晶喷泉C.Crystal Fountain、超级多蓝C.Belleof Taranaki、魔法喷泉C.Magic Fountain、伍斯特美女C.Beauty of Worcester、王梦C.King s Dream、约瑟芬C.Josephine、总统C.The President、丹尼尔德隆达C.Daniel Deronda等品种。期待本研究结果可为铁线莲优良品种的定向选育和市场推广应用提供参考。

Cytotoxic Activities of Total Saponins from Plena Clematis on Human Tumor Cell Lines In Vitro

Objective： To investigate the anti-proliferative effects of saponins prepared from Plena Clematis （PC） cultured in Fujian Province, China on 4 human tumor cell lines and its possible anti-tumor mechanism. Methods： The growth inhibition assays of saponins on human esophageal squamous carcinoma cell line （EC9706）, human hepatoma cell line （HepG-2）, human oral cancer cell line （KB） and human gastric cancer cell line （BGC-823） were evaluated in vitro by thiazolyl blue （MTT） method. The inhibitory effects on EC9706 treated with different concentrations of saponins （15.62, 31.25, 62.50, 125, 250 and 500 μg/mL） were performed in vitro by MTT method. The morphology and nuclear staining with acridine orange/ethidium bromide of EC9706 calls treated with saponins were illustrated under an inverted phase fluorescence microscope. The apoptotic effects of saponins were further evaluated by annexin-V/propidium iodide dual staining experiment to examine the occurrence of phosphatidylserine externalization onto the call surface by a flow cytometer. Results： MTT assay showed that the saponins could inhibit the proliferation of 4 tumor cell lines. Among them, the maximum inhibition rate of 73.1% was detected in EC9706 cells at the saponins concentration of 250 μg/mL for 24 h. Further investigation indicated that the saponins induced EC9706 cells apoposis. The EC9706 cells presented apoptotic characteristics when treated with saponins, including that the morphologies of EC9706 cells were appeared round-shaped with higher refraction, and the cell nuclear stained orange with EB after 250 μ g/mL saponins exposure. The flow cytometry analysis results showed that the induction of cell cycle arrest in apoptotic system may participate in the anti-proliferative activity of saponins on EC9706 cells. Conclusion： The saponins from PC exhibited significant cytotoxicity against human EC9706, KB, BGC-823, and HepG-2 cells and might be beneficial to development of ethnic pharmaceutical plant for potential anti-tumor drugs.

Breeding system and pollination ecology of a potentially invasive alien Clematis vitalba L.in Ireland

Aims Invasive alien plants can greatly affect native communities and eco-system processes but only a small fraction of alien plant species become invasive.Barriers to establishment and invasion include reproductive limitations.Clematis vitalba L.has been a popular hor-ticultural species for the past century and is widely distributed and can be highly invasive.In Ireland,it is considered naturalized and potentially invasive.Despite this,little is known about its reproduc-tive biology.Methods We carried out manipulative field experiments in Ireland and com-pared fruit and seed set from a number of pollination treatments,namely cross-pollination,geitonogamy,autogamy and natural pol-lination.We also recorded floral visitation to C.vitalba through a series of timed observations.Important Findings We found that C.vitalba is capable of uniparental reproduction via geitonogamy and autonomous selfing,albeit at a reduced rate com-pared with outcrossing treatments.Clematis vitalba was visited by at least 10 native pollinator taxa,with hoverflies dominating visitation.Neither fruit set nor seed set in our study population was pollen limited.Given the lack of reproductive constraint,C.vitalba may easily spread in suitable habitats.This is of concern in Ireland,given its prevalence in some of the country’s most floristically diverse regions.

Novel triterpene saponins isolated from Clematis mandshurica and their inhibitory activities on NO production

Four new triterpene saponins, mandshunosides F–I(1–4), together with five known compounds(5–9), were isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of spectroscopic evidences and hydrolysis products. Bisdesmosidic saponin(3–9) showed modest suppression of NO production with the inhibition ratios in the range of 51.3%– 64.6% at 50 μmol·L^(-1), whereas monodesmosidic saponins with a free carboxyl group at C-28(1 and 2) showed potent inhibitory activities with IC_(50) values being 12.7 and 8.3 μmol·L^(-1), respectively.

铁线莲属植物挥发成分的GC-MS研究

In this paper,essential oils from seven different species of Clematis plant were extracted by water steam distillation.Their chromatography fingerprints were obtained by gas chromatography-mass spectrometry (GC-MS).Compounds were mainly identified through mass spectrometry comparing with the NIST02 database.The overlapped peaks were resolved by chemometric resolution methods,obtaining pure chromatography and pure mass spectrometry.The resolved pure mass spectrometry then compared with NIST02 database.The relative quantitative results were obtained by peak area normalization.62 compounds accounting for 87.96% to 97.19% of the total essential oils were identified.It showed that Eugenol,n-hexadecanoic acid and Z,Z-9,12-octadecadienoic acid were their common main compounds although the contents vary with different samples.The chemical compositions in essential oils from different species or different parts of Clematis are also very different.This research provides us chemical bases for further exploitation of Clematis.

粗齿铁线莲扦插繁殖条件初探

为给粗齿铁线莲（Clematis argentilucida）扦插繁殖提供理论依据，进行了影响扦插3个因子的L9（34）正交试验，3个因子分别为基质、生根剂浓度和生根剂处理时间。试验结果表明，粗齿铁线莲扦插生根率可达36.67%~83.30%；生根时间需22~32 d，根数12~23条，根长39.4~87.7 cm。A 2B1C2处理扦插基质为腐殖土：田园土：珍珠岩=3：1：1，外源激素萘乙酸（NAA）浓度为400 mg/L，处理时间10 min的插穗生根情况最优，插穗生根率达83.30%、平均根数22.88条、平均根长87.68 cm、生根所需时间21.20 d，根生长速率107.60μm/d，叶芽生长速率91.30μm/d。同时，主成分分析（PCA）以及隶属函数法分析结果也表明， A 2B1C2处理组合的插条生长情况最好。

铁线莲属药用植物的研究进展

综述了铁线莲属药用植物在各省的研究进展,总结了铁线莲属植物的化学成分及药理作用,并对山东省铁线莲属植物的研究现状进行了研究,为该属资源合理利用提供了依据。

铁线莲属研究随记(VI)(英文)

讨论了Clematis eriopoda Maxim.和sect.Atragenopsis Boiss.的地位,认为这二分类群均应成立;描述了2新种,1新变种;过去长期被归并的卷萼铁线莲C.tubulosa得到恢复;Clematisheracleifoliavar.ichangensis被转移改作卷萼铁线莲的变种;首次给出光叶铁线莲Clematisglabrifolia的果实的形态描述。