Karyological notes on eight species of Achillea L. (Asteraceae, Santolinoideae) from Turkey

The chromosome number and morphology of eight species belonging to the section Santolinoideae of the genus Achillea L. (Asteraceae) were investigated using karyological techniques. The species studied were Achillea cucullata (Hausskn.) Bornm., A. vermicularis Trin., A. monocephala Boiss. & Bal., A. boissieri (Hausskn.) Bornm., A. santolina L., A. gypsicola Hub.-Mor., A. goniocephala Boiss. & Bal., and A. spinulifolia Fenzl ex Boiss. In these species, the chromosome numbers were 2 n  = 18 in A. vermicularis , A. monocephala , and A. boissieri , 2 n  = 18A + 1B in A. spinulifolia , 2 n  = 36 in A. cucullata and A. santolina , and 2 n  = 54 in A. gypsicola and A. goniocephala . All of the chromosomes had median point (M), median region (m), or submedian (sm) centromeres. An increase in asymmetry was not observed in the karyotypes of the species.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 573–580.     

Distribution of phenolic compounds in Middleeuropean taxa of the Achillea millefolium L. aggregate

Achillea millefolium L. s.l. is a cytogenetically, morphologically, and chemically polymorphic aggregate. Besides the sesquiterpenes that possess chemotaxonomic relevance and mediate the antiphlogistic activity, the plant contains phenolic compounds such as dicaffeoylquinic acids and flavonoids causing choleretic and spasmolytic effects. To evaluate their contribution to the chemotaxonomy of European taxa of the A. millefolium group, we developed a SPE-HPLC/UV method that allows quantification of the phenolic constituents in the different taxa. The investigated species displayed differences in the quantitative and qualitative composition of phenolic acids and flavonoids. Hence, they seem to be of chemotaxonomic significance, especially for the distinction of the diploid taxa. Combining the obtained results with the data of the sesquiterpene analyses gives a comprehensive insight into the distribution of those pharmacologically relevant plant constituents in the A. millefolium group.     

Essential oil composition and antimicrobial activity of wild and cultivated Moroccan Achillea ageratum L.: a rare and threatened medicinal species

The essential oils of leaves and flowers of the wild and cultivated Moroccan Achillea ageratum L., a rare and threatened medicinal species, were examined by GC/MS, and their chemical compositions were compared. At least nine components were identified in both wild and cultivated A. ageratum oils, representing more than 95% of the oils. Artemisyl acetate (62.34-78.79%), yomogi alcohol (4.89-12.40%), santolina alcohol (4.86-11.77%), and artemisia alcohol (3.36-7.04%) were the major compounds. Terpene-alcohol proportion was higher in wild A. ageratum than in cultivated A. ageratum. The antibacterial analysis showed that both oils presented high activity against all the studied Gram-positive strains in a range of MIC values from 2.55 to 7.02?mg/ml, but they appeared not effective against the tested Gram-negative ones (MIC values 20.40-41.10?mg/ml). They also exhibited remarkable antifungal activities against Candida species with MIC values ranging from 5.83 to 8.42?mg/ml. From these results, it was concluded that domestication of this threatened medicinal species using clonal propagation did not significantly affect its chemical composition and consequently its antimicrobial properties.     

Germacrane-Type Sesquiterpene Lactones from Achillea millefolium L. and Their Anti-Inflammatory Activity

Six undescribed germacrane-type sesquiterpene lactones, millefoliumons A–F, and two known analogs were isolated from the ethyl acetate fraction of the whole plant of Achillea millefolium L. growing in Xinjiang, China. The structures of these compounds were fully elucidated by their 1D and 2D nuclear magnetic resonance (NMR), and high resolution mass (HR-ESI-MS) spectral data, and comparison with literatures. The absolute configurations of millefoliumons A–F were confirmed by experimental and calculated electronic circular dichroism data (ECD), and ¹³C-NMR calculations and DP4+ probability analysis. All compounds displayed the approximate tendency to inhibit the nitric oxide (NO) release in lipopolysaccharide (LPS)-induced BV2 cells.     

AFLP analyses demonstrate genetic divergence, hybridization, and multiple polyploidization in the evolution of Achillea (Asteraceae-Anthemideae)

Achillea, a temperate genus of herbaceous allogamous perennials, is a model for evolutionary radiation through hybridization and polyploidization. AFLP analyses were performed on 300 individuals of 66 populations and 27 taxa/cytotypes, mainly from the polyploid A. millefolium aggregate and its suspected hybrid links with other clades of the genus. The mosaic genetic structure of hybrids and polyploids is revealed by specific AFLP bands shared with their assumed parents. In E Asia, A. alpina-4x and A. wilsoniana-4x are allotetraploids between A. acuminata-2x (sect. Ptarmica) and A. asiatica-2x (sect. Achillea-A. millefolium agg.). A. virescens-4x is a hybrid species linking A. nobilis agg. and A. millefolium agg. in S Europe. The hybrid swarm A. clypeolata-2x yen A. collina-4x recently formed in Bulgaria shows no AFLP bands additive to its parents; by contrast, other more ancient allopolyploids exhibit genetic innovations. Relationships within A. millefolium agg. are complex. Five 2x-taxa, mostly well separated and regressive, are limited to Eurasia; seven 4x- and 6x-taxa are intimately linked by hybridization, are expansive, and through A. asiatica-2x/4x have formed the N American polyploids. All these results from AFLPs correspond well to other evidence, and indicate a long history of reticulate evolution in Achillea.     

Lectotypification and description of Clematis viticella L. (Ranunculaceae)

The typification of Clematis viticella L. is discussed, and a lectotype designated from the Clifford Herbarium (BM). A standard specimen for C. viticella 'Purpurea Plena Elegans' has also been selected. Data from hybridization experiments and studies of natural distribution suggest that Clematis campaniflora Brotero is better treated as a subspecies, i.e. as C. viticella subsp. campaniflora (Brotero) Font Quer ex O. Bolòs & Vigo, than as either a species or variety.     

Sesquiterpene lactones from Achillea collina J. Becker ex Reichenb

The flower heads of Achillea collina afforded 31 individual sesquiterpene lactones, among which nine guaianolides (1, 16, 20, 21, 23, 27-30), a germacranolide (18) and a dimeric guaianolide (31) were found to be new natural products. The lactones 2-4, 12, 14, 15, 17, 19 and 25 were isolated for the first time from the investigated species. Their structures were established by spectral methods.     

Highly hydroxylated guaianolides of Achillea asiatica and Middle European Achillea species.

From flower heads of Achillea asiatica (L.) Serg., three new guaianolides were isolated by repeated column chromatography and HPLC. The constitution and the stereochemistry of these new, labile compounds were determined by MS, one ((1)H, (13)C, selective (1)H-TOCSY and (1)H-NOESY) and two-dimensional NMR experiments ((1)H, (1)H-COSY, (1)H, (13)C-HSQC, (1)H, (13)C-HMBC). The substances were identified as 8 alpha-angeloxy-2 alpha, 4 alpha,10 beta-trihydroxy-6 beta H,7 alpha H, 11 beta H-1(5)-guaien-12,6 alpha-olide (1), 8 alpha-angeloxy-1 beta,2 beta:4 beta,5 beta-diepoxy-10 beta-hydroxy-6 beta H, 7 alpha H, 11 beta H-12,6 alpha-guaianolide (2) and 8 alpha-angeloxy-4 alpha,10 beta-dihydroxy-2-oxo-6 beta H,7 alpha H, 11 beta H-1(5)-guaien-12,6 alpha-olide (3). They were also detected in Middle European species (Achillea collina, Achillea ceretanica (2x and 4x), Achillea roseoalba, Achillea asplenifolia) by HPLC, TLC and off line MS and have not been described before. The possibility that these compounds might be products of an oxidation process is discussed.     

Variation in Terpene and Linear‐Chain Hydrocarbon Content in Yarrow (Achillea millefolium L.) Germplasm from the Rhaetian Alps,Italy

Yarrow (Achillea millefolium L.) is a herbaceous species common in the Alpine region of Europe and used in folk medicine since antiquity. Its organs are rich in monoterpenes and sesquiterpenes, two subclasses of plant terpenoids with relevant ecological significance, which were reported as valuable markers for the traceability of mountain dairy products. The variability in chemical composition of yarrow germplasm may be related with its genetic diversity, accounting for possible differences in medical properties, and supporting its use as a specific territorial marker. Aim of this work was to assess the leaf chemical composition of 16 yarrow populations collected at altitudes exceeding 1600 m in three valleys of the Rhaetian Alps, Italy, and jointly evaluated in a lowland site. The most abundant compounds detected generally differed from those of the germplasm from other countries. A trend of valley‐specific pattern of composition was evident. However, the variability among individual populations was even more remarkable, regardless of their valley of origin. The concentrations of sesquiterpene hydrocarbons, oxygenated monoterpenes, and oxygenated sesquiterpenes discriminated the populations in multivariate analysis. A few prevailing chemotypes were characterized, which differed from those previously reported in the literature. The geographic isolation from other germplasms, and the local ecotypization, likely originated a chemically distinct gene pool.     

蓍草化学成分研究

研究蓍草Achillea alpine L.全草的化学成分。采用大孔树脂、ODS、Sephadex LH-20凝胶柱色谱和pre-HPLC等方法分离与纯化,运用NMR、MS等波谱技术鉴定化合物结构。从蓍草95%乙醇提取物中分离得到16个化合物,分别是(2 E,4 E)-N-(2-methylbutyl)deca-2,4-dienamide(1)、墙草碱(2)、(E,E,Z)-2,4,8-decatrienoicacid isobutylamide-8,9-dehydropellitorine(3)、N-2′-methylbutyl-(E,E)-2,4-decadienam(4)、methyl-(E,E)-2,4,9-oxooctadeca-10,12-dienoate(5)、(S)-14-(E,E)-10,12-methyl 14-hydroxy-9-oxo-octadeca-10,12-dienoate(6)、(E,E)-2,4-undecadiene-8,10-diynamide-N-(2-methylpropyl)(7)、(E,E)-2,4-decadienoic acid p-hydroxyphenethylamide(8)、sinapyl alcohol diisovalerate(9)、(S)-13-hydroxyoctadeca-(Z,E)-9,11-dienoic acid(10)、(E,E)-2,4-decadienamide acid p-methoxyphenethylamide(11)、erythro-N-isobutyl-4,5-dihydroxy-2-(E)-decenamide(12)、3-O-阿魏酰-奎宁酸(13)、肉桂酸(14)、绿原酸(15)、3-O-咖啡酰-5-O-阿魏酰奎宁酸(16)。化合物1是一个新的酰胺类化合物;化合物4~6、9、10、12、13、16为首次从该属植物中分离得到;化合物8、11为首次从蓍草中分离得到。化合物1~11在四种不同的胃癌细胞株上进行细胞毒活性筛选,结果显示化合物2、5与9在50μM时对MGC-803细胞株具有较弱抑制活性,其抑制率依次为38.7%、34.7%、31.5%。     

Hybrid origin and differentiation of two tetraploid Achillea species in East Asia: molecular, morphological and ecogeographical evidence

Achillea (Asteraceae-Anthemideae) offers classical models for speciation by hybridization and polyploidy. Here, we test the suspected allotetraploid origin of two species, Achillea alpina and Achillea wilsoniana between phylogenetically distinct lineages in East Asia. A total of 421 AFLP bands from 169 individuals and 19 populations of five 2x- and two 4x-species were obtained. The data set was analysed with a newly developed model that accounts for polyploidy and assumes lack of recombination between the parental chromosome sets (i.e. disomic inheritance). A. alpina and A. wilsoniana then appear to be allotetraploids between Achillea acuminata-2x (sect. Ptarmica) and Achillea asiatica-2x (sect. Achillea). The two 4x-species share 44% and 48% of their AFLP bands with A. acuminata-2x, and 39% and 38% with A. asiatica-2x, respectively. Eight plastid haplotypes (A-H) were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses. A. alpina-4x and A. wilsoniana-4x share haplotype F only with A. asiatica-2x. This is consistent with the hybrid origin(s) involving the latter as the maternal ancestor. This result corroborates our previous DNA sequence data, where A. alpina-4x and A. wilsoniana-4x are also placed close to A. asiatica-2x. Morphology, ecology, and amplified fragment length polymorphism (AFLP) profiles of the two 2x-species are distinct, whereas the two 4x-species, grouped as A. alpina aggregate, form a nearly continuous link between them. Considering all evidence, this 4x-aggregate is regarded as the product of a hybridization between genetically distant 2x-ancestors limited to China and adjacent areas: one A. acuminata-like, and the other A. asiatica-like. The allopolyploid A. alpina agg. exhibits considerable morphological variation and ecological flexibility, and has expanded throughout eastern Asia and to northern North America, far beyond the ranges of their presumed 2x-ancestors.     

Antioxidant and Antimelanogenic Activities of Compounds Isolated from the Aerial Parts of Achillea alpina L.

Achillea alpina is widely distributed in Korea and is often used as a folk medicine for stomach disorders. Although a previous study isolated antioxidant compounds (flavonoid O‐glucoside, sesquiterpene) from this plant, no systematic study of its chemical constituents had been reported. The present study aimed to identify the phytochemicals present in a methanol extract of A. alpina, assess their potential antioxidant activities in vitro, and determine their effects on melanogenesis in B16F10 melanoma cells. Column chromatographic separation of aqueous fractions of A. alpina led to the isolation of 17 compounds. The chemical structures of these compounds were determined using spectroscopic data from electrospray ionization‐mass spectrometry and nuclear magnetic resonance. To the best of our knowledge, the present study is the first to identify compounds 2 – 10 and 12 – 17 in A. alpina. Furthermore, compound 6 possessed powerful antioxidant activity, while compound 15 suppressed intracellular tyrosinase activity and thus reduced melanogenesis in B16F10 cells. Therefore, our research suggested that these naturally occurring compounds have the potential to reduce oxidative stress and promote skin whitening. Further investigations will be required to elucidate the mechanisms underlying the antioxidant and antityrosinase activities of these compounds.     

Achillea filipendulina Lam.: Chemical Constituents and Antimicrobial Activities of Essential Oil of Stem,Leaf, and Flower

In this study, we extracted the essential oils of the stem, leaf, and flower of Achillea filipendulina, analyzed them, and studied their antibacterial properties. Of 16, 53, and 35 compounds identified in the stem, leaf, and flowers, respectively, only five are present in all three segments of the plant. The essential oil of the stem was mainly composed of neryl acetate, spathulenol, carvacrol, santolina alcohol, and trans‐caryophyllene oxide. However, the main identified components of leaf were 1,8‐cineole, camphor, ascaridole, trans‐isoascaridole, and piperitone oxide and the main components of the flower oil were ascaridole, trans‐isoascaridole, 1,8‐cineole, p‐cymene, and camphor. The extracted oil from different segments demonstrated varying antibacterial properties against both Gram‐positive and Gram‐negative bacteria, demonstrated by disk, minimum inhibitory concentration, and minimum bactericidal concentration methods. These suggest that the application of all segments of aerial parts of A. filipendulina may have a better therapeutic effect in fighting pathogenic systems.     

Infrared spectroscopy as a tool for chemotaxonomic investigations within the Achillea millefolium group

An FT-IR-based method for the rapid characterization and identification of Achillea species was developed. With the IR spectra of the CH2Cl2 extracts of A. aspleniifolia, A. collina, A. millefolium, A. millefolium ssp. sudetica, A. pannonica, A. pratensis, A. roseoalba, A. setacea, and A. styriaca, a spectrum library was created. This database allowed the identification of the respective taxa by comparison of IR spectra. Moreover, information concerning dominant structural elements of sesquiterpene constituents was obtained, providing hints about the composition of a plant sample of unknown origin.     

Sesquiterpene lactones from Achillea chrysocoma and Achillea coarctata

The aerial parts of A. chrysocoma and A. coarctata afforded 22 and 9 sesquiterpene lactones, respectively. All the compounds described are new for the studied species. The lactones 5, 6 and 12 are new for the genus Achillea.     

Exudate flavonoid aglycones in the alpine species of Achillea sect. Ptarmica: Chemosystematics of A. moschata and related species (Compositae-Anthemideae)

In completion of our studies on Achillea exudate flavonoids, 11 alpine species of Achillea sect. Ptarmica were analyzed for their aglycone profiles. The study focuses on species commonly associated with A. moschata. The major flavonoid constituents found in exudates of most taxa were 6-hydroxyflavonol 3,6,4'-trimethyl ethers, except in A. ageratifolia and its subspecies, which are characterized by the accumulation of the 3,6,7-trimethoxy and 6-hydroxyflavones. Infraspecific variation was particularly high in A. abrotanoides and A. moschata. Results are discussed in relation to published data for related species and within the context of evolutionary aspects in the genus Achillea. The 3,6,4'-trimethoxy substitution is regarded as a basic chemical trend within the genus Achillea. Geographical and ecological aspects are briefly addressed, and a summary on known exudate aglycone composition of species from all sections of Achillea is included.     

Phenolic compounds from Achillea millefolium L. and their bioactivity

Since antiquity, Achillea millefolium L. (Asteraceae) has been used in traditional medicine of several cultures, from Europe to Asia. Its richness in bioactive compounds contributes to a wide range of medicinal properties. In this study, we assessed A. millefolium methanolic extract and its isolated components for free radical scavenging activity against 2,2-diphenyl-pycrilhydrazyl, total antioxidant capacity (based on the reduction of Cu(++) to Cu(+)), and ability to inhibit lipid peroxidation. The activity against chloroquine-sensitive and chloroquine-resistant strains of Plasmodium falciparum was also tested. Chlorogenic acid, its derivatives and some flavonoids isolated by semipreparative HPLC and identified by NMR and spectrometric techniques were the major bioactive constituents of the methanolic extract. The latter exhibited significant antioxidant properties, as well as its flavonol glycosides and chlorogenic acids. With regard to the antiplasmodial activity, apigenin 7-glucoside was the most effective compound, followed by luteolin 7-glucoside, whereas chlorogenic acids were completely inactive. On the whole, our results confirmed A. millefolium as an important source of bioactive metabolites, justifying its pharmaceutical and ethnobotanical use.     

A new species of Achillea (Asteraceae: Anthemideae) from south-east Peloponnisos, Greece

A new species of Achillea L., Achillea occulta Constantin. & Kalpoutz., from the summit area of Mount Koulochera in south-east Peloponnisos, Greece, is described and illustrated. It belongs to A . sect. Ptarmica and further, to a small group of taxa known under the illegitimate sectional name 'Anthemoideae'. In Greece, A. occulta has no close allies. The taxonomically related taxa, i.e. A. barrelieri (Ten.) Sch. Bip., A. mucronulata Bertol., A. oxyloba (DC.) Sch. Bip. and A. schurii Sch. Bip. are found in the Italian Peninsula, the Alps and the Carpathians. Phytogeographically, the finding of a species with such taxonomic connections in south-east Greece was unexpected. The new species grows in semi-shade, often hidden in suitable limestone rock hollows and the foot of rocks, together with several other Greek endemics. Its chromosome number of 2 n  = 18, counted in root tips, is also reported and illustrated.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 249–256.     

Achillicin,the first proazulene from Achillea millefolium

Achillicin, the major proazulene (prochamazulene) of Achillea millefolium L. ssp. collina Becker has been isolated and identified as 8-acetoxyartabsin. It is the first proazulene reported for the genus Achillea and has not been found previously in plants.     

Achillinin A,a Cytotoxic Guaianolide from the Flower of Yarrow,Achillea millefolium

Achillinin A (2β,3β-epoxy-1α,4β,10α-trihydroxyguai-11(13)-en-12,6α-olide, 1), a new guaianolide isolated from the flower of Achillea millefolium, exhibited potential antiproliferative activity to A549, RERF-LC-kj and QG-90 cells with 50% inhibitory concentration (IC₅₀) values of 5.8, 10 and 0.31 μM, respectively.