Genus Clematis Research Articles

Диагностика пораженности клематисов мучнистой росойс помощью методов сканирующей электронной и конфокальной лазерной микроскопии

Приведены результаты мониторинга распространения и интенсивности развития мучнисто-росяного патогена в коллекции клематисов отдела декоративных растений Главного ботанического сада им. Н. В. Цицина РАН, выполненного методами сканирующей электронной и конфокальной лазерной микроскопии. Установлено поражение 13-ти сортов клематиса 6-ти садовых групп выявленным грибом Erysiphe aquilegiae var. ranunculi (Grev.) R.Y. Zheng& G.Q. Chen. The article is devoted to the investigation of the powdery mildew pathogens in the collection of Clematis L. in the Department of Ornamental Plants of the Main Botanical Garden named after N.V. Tsitsin RAS. In the first part of the article, the authors present general information about these ornamental plants. The taxonomy of mildew fungi parasitizing on the genus Clematis L. is discussed. A historical reference and modern assessment of the pathogen species belonging are given and the danger of the development of this disease in the collections is presented. In the next part of the work, the botanical composition of the studied collection is presented, and the methods and techniques is described, especially scanning electron and confocal laser microscopy, as well as morphometric analysis. In the section devoted to the results of the study, an assessment is made of the degree of damage to the plants of the clematis collection and a detailed morphometric characterization of the identified mildew pathogen, based on the microphotographs obtained by the cryoSEM and KLSM methods. After analyzing the morphometric data, the authors summarize their work and conclude that a collection of the genus Clematis L. has been infested with Erysiphe aquilegiae var. ranunculi (Grev.) R.Y. Zheng & G.Q. Chen. It is concluded, that plant damage as a wholew a not high, because the pathogen was found only in 13 from 60 examined Clematis species visually and by the cryo-SEM method. Severe and medium damage by the mildew fungus E. aquilegiae var. ranunculi was registered in 4 representatives of the collection. A minor damage was registered in 9 varieties of the collection.

Chromosome count and karyotype of two species of Clematis (Ranunculaceae)

Karyotype morphometric data of two species of the genus Clematis L., a cosmopolitan and diverse genus, were studied. Clematis flammula is a diploid with 2n = 16 chromosomes. It exhibits a gradual decrease in size of chromosomes from 14.83 (±3.6) µm to 7.58 (±3.65) µm. Its karyotype formula (2n = 2x = 16 = 10m + 6sm) differs from those of Clematis orientalis (2n = 4x = 32 = 18m + 8sm + 6st), which is in addition a tetraploid species. The intrachromosomal asymmetry (A1) indicates that C. flammula had the most symmetric karyotype A1 = 0.39 in comparison to C. orientalis (A1 = 0.96). The karyological data available in the literature for the genus have also been discussed against the backdrop of the present analysis. This is the first attempt to present the karyotype morphometric data for the tetraploid cytotype of C. orientalis from India.

Seasonal Development of Genus Clematis L. Representatives in Kyiv Conditions

Seasonal Development of Genus Clematis L. Representatives in Kyiv Conditions

A new taraxastane-type triterpenoid glycoside from the roots of Clematis uncinata

A new taraxastane-type triterpenoid glycoside, clematiunicinoside I (1), together with four known ones (2–5), was isolated from the roots of Clematis uncinata. The structure of the new compound was elucidated on the basis of spectroscopic analyses and acid hydrolysis. The cytotoxic activities of all the compounds against caski cervical cancer (Caski) cells were evaluated. This is the first report of the presence of taraxastane-type triterpenoid glycoside in the genus Clematis.

Clematis diebuensis, a new specis of Ranunculaceae from Gansu

A species of the genus Clematis sect. Atragene( Ranunculaceae),C. diebuensis,is described as new from Gansu Province. The morphological differences between it and its ally and a smilar species are given.

Metabolic Profiling of Clematis species to identify compounds with an inhibitor effect on NF-κB transactivation

Clematis L. (Ranunculaceae) is a genus of about 300 species widespread throughout the world. Many Clematis species are traditionally used for various ailments, among them many diseases related to inflammation. Based on this knowledge and on the results of a large screening of Chinese medicinal plant extracts, the genus Clematis was chosen as a subject for a study in order to explore the feasibility of correlating metabolic profiles and bioactivity of crude extracts to identify their active constituents. Ethanolic extracts from 48 different Clematis samples originating from C. uncinata, C. smilacifolia, C. finetiana, C. gouriana, C. leschenaultiana, C. chingii, C. loureiroana, C. apiifolia var. argentilucida, C. chinensis, C. meyeniana, C. peterae, C. parviloba, C. puberula var. tenuisepala and C. buchananiana were included in this investigation. The extracts were tested for inhibition of NF-κB luciferase reporter transactivation in HEK293 cells [1]. At a concentration of 50 µg/ml, the extracts showed distinct activities, some of them possessing a very good inhibitory effect. In parallel, the metabolic profiles of the extracts were analysed by LC-HRMS in the ESI negative mode (Dionex Ultimate 3000 UHPLC; Q ExactiveTM Hybrid Quadrupole Orbitrap-MS, Thermo Fisher). The LC-MS data were processed in an untargeted approach by MZmine 2 [2]; peaks were verified and quantified by Lipid Data Analyzer [3]. The abundance of the peaks was linked to the pharmacological activity of the extracts using the software SIMCA 13 [4]. Principal component analyses as well as orthogonal partial least squares-discriminant analyses (OPLS-DA) and visualisation of the data by means of S-plot was conducted. This approach led to the identification of eight phenolic compounds which seem to be highly relevant for NF-κB inhibitory activity.

Karyotype analysis on 11 species of the genus Clematis

In order to explore the karyotype and evolutionary trend of Clematis and provide taxonomically useful data, the chromosome number, the number and position of satellites, the karyotype formulae, the karyotype, arm ratio, relative length centromeric index, and the index of the karyotype asymmetry of 11 species of the genus Clematis were studied. All the analyzed species showed the same stable chromosome number (2n = 2x = 16) and basic chromosome number 8. Clematis karyotypes are composed by metacentric, submetacentric, and terminal centromeric chromosomes. Clematis lanuginosa, C. otophora, C. lasiandra, C. shenlungchiaensis, C. florida var. plena, C. fusca var. violacea, C. crispa, and C. viticella belong to Type 2A, a very symmetric and plesiomorphic type. Clematis henryi, C. integrifolia, and C. japonaca belong to Type 2B, which is apomorphic relative to the other group. The 11 species of Clematis are different in karyotype parameters providing data for the taxonomy of the genus.

Three new glycosides from the whole plant of Clematis lasiandra Maxim and their cytotoxicity

Phytochemical investigation on the whole plant of Clematis lasiandra Maxim led to the isolation of two new phenolic glycosides (1 and 2), one new lignanoid glycoside (3), together with three known lignanoid glycosides (4–6). The structures of the new compounds were elucidated as 4-O-β-d-galactopyranosyl-ethyl-E-caffeate (1), 4-O-β-d-galactopyranosyl-3-hydroxyl-phenylethene (2) and (8R)-3,3′-dimethoxy-4,4′,9,9′-tetrahydroxy-5′,8-lignan 3′-O-β-d-glucopyranoside (3), on the basis of extensive spectral analysis and chemical evidence. The characteristic of the polymerized C-5′–C-8 type lignanoid aglycone in glycoside 3 was found from genus Clematis for the first time. Compounds 1–6 were evaluated for their cytotoxicity against human tumor cell lines HL-60, Hep-G2 and SGC-7901, the new glycosides 1 and 2 showed significant cytotoxicity against those three tumor cell lines with IC50 in the range from 9.73 to 22.31μM, while lignanoid glycosides 3–6 showed weak cytotoxicity to those test cell lines with IC50 value more than 52.71μM.

Triterpenoid saponins from the roots of Clematis argentilucida

Reinvestigation of the n-BuOH extract of the roots of Clematis argentilucida led to the isolation of a new ursane-type triterpenoid saponin 1 and a new taraxerane-type saponin 2, four known saponins 3–6 first isolated from the species, together with seven saponins 7–13 reported in the previous papers. The structures of saponins 1–6 were elucidated by extensive spectroscopic analysis and chemical evidences. The ursane-type and taraxerane-type triterpenoid saponins were obtained from genus Clematis for the first time, and the aglycone of saponin 1, 3β,28-dihydroxy-18αH-ursan-20-en was first encountered. The cytotoxicity of all the saponins was evaluated against human glioblastoma U251MG cell lines. The monodesmosidic saponins 1, 2 and 4–8 exhibited cytotoxic activity against the cells with IC50 values ranging from 6.95 to 38.51μM.

Two new names in Clematis (Ranunculaceae)

Molecular studies (Miikeda et al. 2006, Xie et al. 2011) have established that the genus Naravelia Adanson (1763: 460) nom. cons. is nested within the genus Clematis Linnaeus (1753: 543). Most of the names treated under Naravelia have combinations in Clematis already. However, N. siamensis Craib and N. pauciflora H.Eichler have not been transferred to Clematis, apparently because of the existing homonyms. Here, we provide new names for them.

A new species of Clematis L. (Ranunculaceae) from Iran

After extensive sampling and investigations of the genus Clematis from various locations in Iran, Clematis iranica is introduced as a new species. The diagnosis, description and illustration of the new species are presented here. The affinities and differences to the most closely related species (C. orientalis) are presented based on morphological, cytological, palynological and anatomical characters.

The genus Clematis (Ranunculaceae): Chemical and pharmacological perspectives

Ethnopharmacological relevanceTwenty six species of the genus Clematis (Ranunculaceae) have been traditionally used in various systems of medicine for the treatment of ailments such as nervous disorders, syphilis, gout, malaria, dysentry, rheumatism, asthma, and as analgesic, anti-inflammatory, diuretic, antitumour, antibacterial and anticancer. Aim of the reviewTo emphasize on ethnopharmacology, chemical constituents, pharmacology, toxicology and clinical studies of various species of the genus Clematis. Materials and methodsThe available information on Clematis species was collected through electronic search of major scientific databases. ResultsA survey of literature revealed that triterpene saponins, alkaloids, flavonoids, lignans, steroids, coumarins, macrocyclic compounds, phenolic glycosides, anemonin and volatile oils constitute major classes of chemical constituents in the genus Clematis. Preliminary analgesic, anticancer, anti-inflammatory, diuretic, antiarthritis, hepatoprotective, hypotensive and HIV-1 protease inhibitor activity studies have been carried out on crude extracts of 26 traditionally used and medicinally promising species of Clematis genus. ConclusionsThe species of the genus Clematis emerged as good source of traditional medicine for the treatment of various ailments. Although few experimental studies validated their traditional claims, but employed uncharacterized crude extracts. Such Clematis species need to be explored properly following bioactivity-directed fractionation with a view to isolate bioactive constituents, and to evaluate their possible mode of actions. These species hold great potential for detailed clinical studies so that these could be exploited as potential drugs. The review will help researchers to select medicinally potential species of Clematis genus for future research.

Two new cytotoxic triterpenoid saponins from the roots of Clematis argentilucida

Bioassay-guided fractionation of the n-BuOH extract of the roots of Clematis argentilucida led to the isolation of two new triterpenoid saponins along with a known one, cussonside B (3). By extensive spectral analysis and chemical evidences, the structures of the two new saponins were determined to be 3β-O-[β-d-ribopyranosyl-(1→3)-α-l-rhamnopyranosyl-(1→2)-α-l-arabinopyranosyl] hederagenin-11,13-dien-28-oic acid (1) and 3β-O-{β-d-ribopyranosyl-(1→3)-α-l-rhamnopyranosyl-(1→2)-[β-d-glucopyranosyl-(1→4)]-β-d-xylopyranosyl} oleanolic acid (2), respectively. Saponin 1 is the first example of triterpenoid saponins with two double bonds located at C-11 and C-13 in the aglycone from the genus Clematis. The two new saponins exhibited significant cytotoxicity against human leukemia HL-60 cell lines, human hepatocellular carcinoma Hep-G2 cell lines and human glioblastoma U251MG cell lines with a range of IC50 values from 2.74 to 25.40μM, while 3 showed inactivity against all of the three cancer cell lines.

Distribution of the triterpenoid saponins and chemotaxonomy of the genus Clematis L. by high-performance liquid chromatography–mass spectrometry

Selected species of the genus Clematis (Ranunculaceae) have been screened for occurrence of triterpenoid saponins by qualitative HPLC-MS n analysis of root and rhizome materials from 18 Clematis samples as well as the whole plant materials of Clematis puberula var. ganpiniana and Clematis terniflora. The HPLC-MS n analysis allowing the detection of 17 oleanolic acid or hederagenin saponins was carried out in the negative selected ion monitoring (SIM) mode. Triterpenoid saponin profiles of these taxa were used for phylogenetic studies, and results are presented as a dendrogram. Huzhangoside B could be unambiguously identified in all analyzed Clematis taxa, as well as in the investigated Ranunculus taxa. The distribution and chemotaxonomic importance of the triterpenoid saponin profile within this genus are discussed.

A comparative study using chemometric and numerical taxonomic approaches in the identification and classification of traditional Chinese medicines of the genus Clematis

A comparative study was performed to identify and classify traditional Chinese medicines (TCMs) of the genus Clematis by applying chemometric and numerical methodologies. In the analysis, 12 species representing major TCM plants of the genus and belonging to five different sections – Rectae, Clematis, Meclatis, Tubulosae and Viorna – were identified and demarcated on the basis of morphological and phytochemical characters. In the numerical study (NS), out of 53 selected characters, only 27 phylogenetically informative characters were used for data generation; a phenogram was produced with three distinctive clades. The extent of genetic distances ranged from 0.35 to 0.80. The phytochemical analysis (PA) was conducted using a high performance liquid chromatography coupled with a diode array detector (DAD) and ESI‐MS mode. The compound “Huzhangoside D” (HGD) was the most abundant in the analysed species of the genus. The PA of saponins produced a differential matrix based on the presence or absence of chemical characters, which generated a phenogram. The extent of genetic distances ranged from 0.05 to 0.35 in the phytochemical analysis, with three distinctive clades. Clematis intricata had compounds (HGB, HGD) with GD 0.30, and produced a different pattern of clustering than the previous classification system. The species C. henryi and C. heracleifolia appeared as sister clades having congruent genetic similarity with each other; this is an innovative combination. The classification results of PA and NS approaches corroborate each other, but the chemical fingerprinting method proved more authentic, quick and reliable for the identification and classification of these plant taxa.

Characterization and identification of triterpenoid saponins in crude extracts from Clematis spp. by high‐performance liquid chromatography/electrospray ionization with multi‐stage tandem mass spectrometry

High-performance liquid chromatography coupled with electrospray ionization multi-stage tandem mass spectrometry (HPLC/ESI-MS(n)) was applied to characterize and identify triterpenoid saponins in crude extracts from nine species of Clematis L. After separation on a Zorbax SB-C(18) column, negative ESI-MS experiments were performed. The quasi-molecular ions [M-H]-, [M+Cl]- and [M+HCOO]- were observed in the full-scan MS spectra of all compounds. The MS(n) (n = 2-4) data of the [M-H]- ions provided structural information on the sugar sequence of the oligosaccharide chains and on the aglycone of the saponins. In addition, the fragmentation mechanisms could be deduced from the fragment ions. As a result, eight saponins were unambiguously identified in C. ganpiniana by comparison with reference compounds. In addition, a new compound was tentatively identified as 3-O-ribopyranosyl --> rhamnopyranosyl --> (glucopyranosyl) --> arabinopyranosylhederagenin 28-O-rhamnopyranosyl --> glucopyranosyl --> glucopyranosyl ester (peak 1), and another one was tentatively deduced to be 3-O-glucopyranosyl --> ribopyranosyl --> rhamnopyranosyl --> arabinopyranosylhederagenin 28-O-rhamnopyranosyl --> glucopyranosyl --> glucopyranosyl ester (peak 5) from the genus Clematis L. for the first time. By ESI-MS(n), non-isomeric saponins could be discriminated rapidly. It is of interest that cleavage preferentially occurrs at the ester bond at C-28 and the charge is easy to transfer onto the oligosaccharide chain when the ester bond of a monodesmosidic saponin like HNH cleaves.

First Report of Leaf Spot Caused by a Phoma sp. on Clematis × jackmanii in Italy.

The genus Clematis, belonging to the Ranunculaceae family, is widely used in gardens and is very much appreciated for its climbing attitude as well as rich flower production. In the fall of 2006, in a private garden located near Biella (northern Italy), a severe foliar disease was observed on 2-year-old plants of Clematis × jackmanii. Small necrotic spots were observed on the upper and lower sides of infected leaves. At temperatures of 15 to 25°C, spots enlarged to form round areas that were 2 to 7 cm in diameter and well defined by a brown margin. Severely infected leaves wilted without abscising. The disease occurred on 100% of the plants of the C. × jackmanii hybrid in one garden. Stems and flowers were not affected by the disease. From infected leaves, a fungus was consistently isolated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin. The fungus was grown on PDA and maintained at 22°C (12-h light, 12-h dark). After 10 days, black pycnidia 132 to 340 μm in diameter developed, releasing abundant hyaline, elliptical, nonseptate, conidia measuring 5.1 to 8.3 (6.8) × 1.6 to 3.4 (2.7) μm. On the basis of its morphological characteristics, the fungus was identified as a Phoma sp. (2). The internal transcribed spacer region of rDNA was amplified using primers ITS4/ITS6 (1,3), sequenced (GenBank Accession No. EF566917), and identified as a Phoma sp. Pathogenicity tests were performed by spraying leaves of healthy 1-year-old potted C. × jackmanii (cvs. Superba, Mrs N. Thomson, and Vagebond) plants with a spore and mycelial suspension (4 × 105 spores or mycelial fragments per ml). Noninoculated plants served as controls. Five plants per cultivar were used for each treatment. Plants were covered with plastic bags for 3 days after inoculation and kept in a growth chamber at 18 to 20°C. Symptoms previously described developed on leaves of all tested cultivars 10 days after inoculation, while control plants remained healthy. On the infected leaves, pycnidia and conidia with the same dimensions and characteristics as previously described were observed. The fungus was consistently reisolated from the lesions of the inoculated plants. The pathogenicity test was carried out twice. The presence of Ascochyta clematidina, then renamed as Phoma clematidina, on Clematis species has been reported in the United States (4) and subsequently in the Netherlands, Britain, and New Zealand.

Phylogenetic relationships of Clematis (Ranunculaceae) based on chloroplast and nuclear DNA sequences

Phylogenetic relationships within Clematis, including Naravelia, Archiclematis and Clematopsis, were analysed using nucleotide sequences of chloroplast DNA [(1) matK and trnK introns; (2) atpB–rbcL spacer; (3) rpoB–trnC spacer; (4) psbA–trnH–trnQ spacer; (5) rbcL–accD spacer] and the nuclear ITS regions. The phylogenetic trees resulting from these analyses suggested nine major clades. The genera Archiclematis, Naravelia and Clematopsis were found to be nested within the genus Clematis, and should be included within it. Within the genus Clematis, the traditional subgenus Viorna was found to be monophyletic. The remaining subgenera (Flammula, Clematis and Campanella) and sections Flammula, Clematis and Cheiropsis, however, were found to be paraphyletic or polyphyletic. Recircumscription of several groups in Clematis was suggested. Enormous morphological diversification and very few nucleotide substitutions within Clematis indicate the recent radiation of the genus. Clematidinae shares a gene order with Anemoninae between the starting point of large single copy region and trnQ of chloroplast DNA, although Hepatica, a genus in Anemoninae, shows a different order in the region near trnH.

Intersimple Sequence Repeat Fingerprinting and Genetic Variation in a Collection of Clematis Cultivars and Commercial Germplasm

The genus Clematis contains many well-known large flowered cultivars, as well as lesser-known nonvining species. Intersimple sequence repeat (ISSR) primers were used to fingerprint 32 vining cultivars and five nonvining species (C. fruiticosa, C. integrifolia, C. heracleifolia, C. hexapetala, and C. recta) for use in assessing genetic relationships and cultivar identification. Four ISSR primers yielded a total of 44 bands in the vining accessions, of which 36 (86%) were polymorphic. The average polymorphism levels were 83% for the cultivars and 94% for the nonvining species. All 32 vining cultivars were distinguished with the use of two ISSR primers, and the five nonvining Clematis species were differentiated with three ISSR primers. A similarity matrix of the cultivars showed low similarity levels between the samples, with an average similarity of 0.28. A UPGMA-derived dendrogram showed no strong groupings among any of the samples. Two cultivars with known parentage, Clematis viticella L. `Betty Corning' and `Sylvia Denny', grouped with one reported parent but not the other, suggesting they are more similar to one parent. `Multi-blue', a sport reportedly arising from `The President' did not segregate near `The President'.

A classification of genus Clematis

A classification of genus Clematis