Fritillaria Species Research Articles

Taxonomic relationships in genusFritillaria(Liliaceae): Evidence from RAPD-PCR and SDS-PAGE of seed proteins

The genus Fritillaria L. is one of the examples from the several unsolved taxonomic problems. The classical identification of plant depends on morphological characteristics. Therefore, it is difficult to determine the botanical origin of some plants. The goals of this study were to examine the taxonomic status of these 42 taxa of Fritillaria in Turkey by means of RAPD-PCR and seed protein analysis in addition to taxonomic interpretation of species. SDS-PAGE and RAPD-PCR techniques were used to help taxonomic interpretation of Fritillaria species. Dendrogram based on g enetic distances was calculated using the computer programmed PopGen. For the RAPD-PCR analysis, 40 random primers were tested in the amplification reactions with Fritillaria species. Only 9 of them gave consistently reproducible banding patterns. The analysis of seed proteins showed that all studied genotypes had a specific protein pattern except the species that were close to each other based on morphological data. In electrophoretic prote...

Characterizing distribution of steroidal alkaloids in Fritillaria spp. and related compound formulas by liquid chromatography–mass spectrometry combined with hierarchial cluster analysis

Bulbus Fritillariae ( BF), referred to the bulbs of several Fritillaria species (Liliaceae), is a commonly used antitussive and expectorant herb in traditional Chinese medicine (TCM). Due to the complexity of BF botanical origin in the herbal markets, it is urgently needed to develop a reliable method for species identification. Previous studies based on morphological and histological as well as molecular biological techniques have respective limitations. For the purpose of finding a possible discriminant method among Fritillaria species, 27 steroidal alkaloids in 17 Fritillaria species and 12 BF-containing compound formulas were identified and characterized by a high-performance liquid chromatography with mass spectrometry (HPLC–MS) method. The estimated relative composition of steroidal alkaloids was used to carry out a chemotaxonomical study on Fritillaria species by means of hierarchial cluster analysis. In addition, the characteristic occurring patterns of the examined bases were compared in an effort to distinguish the botanical origin of BF-containing compound formulas. The results demonstrated that the qualitative and quantitative differences in steroidal alkaloids were useful not only for chemotaxonomy in some medicinal Fritillaria species but also for species identification in compound formulas. The described method has important implications in quality control of BF-containing TCM preparations, allowing for the prevention of BF confusion, and also revealing the possible presence of adulteration.

Development and validation of a liquid chromatography/electrospray ionization time-of-flight mass spectrometry method for relative and absolute quantification of steroidal alkaloids in Fritillaria species

Steroidal alkaloids are naturally occurring nitrogen-containing compounds in many edible or medicinal plants, such as potato, tomato, Fritillaria and American hellebore, which possess a variety of toxicological and pharmacological effects on humans. The aim of this study is to explore the potential of liquid chromatography/electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF-MS) method in the determination of these important alkaloids in plant matrices. The application of this method has been proven through 26 naturally occurring steroidal alkaloids in Fritillaria species. Accurate mass measurements within 4 ppm error were obtained for all the alkaloids detected out of various plant matrices, which allowed an unequivocal identification of the target steroidal alkaloids. The bunching factor for mass spectrometer, an important parameter significantly affecting the precision and accuracy of quantitative method, was firstly optimized in this work and satisfactory precision and linearity were achieved by the optimization of that parameter. The ranges of RSD values of intra-day and inter-day variability for all alkaloids were decreased remarkably from 41.8–159% and 13.2–140% to 0.32–7.98% and 2.37–16.1%, respectively, when the value of bunching factor was optimized from 1 to 3. Linearity of response more than two orders of magnitude was also demonstrated (regression coefficient >0.99). The LC/TOF-MS detection method offered improvements to the sensitivity, compared with previously applied LC (or GC) methods, with limits of detection down to 0.0014–0.0335 μg/ml. The results in this paper illustrate the robustness and applicability of LC/TOF-MS for steroidal alkaloids analysis in plant samples. In addition, relative quantitative determination of steroidal alkaloid with one popular analyte verticinone which is commercially available was also investigated in order to break through the choke point of lack of standards in phytochemical analysis. The accuracies of relative quantitative method for steroidal alkaloids determinations with verticinone were 90.6–110.0% (average 98.5%) suggesting that it is feasible to quantify steroidal alkaloids by the proposed relative quantitative determination method within acceptable errors.

Modern state and structure of coenotic populations of species of Fritillaria L. genus in some sites of the Poltava region

Modern state and structure of coenotic populations of species of Fritillaria L. genus in some sites of the Poltava region

Simultaneous identification of Bulbus Fritillariae cirrhosae using PCR-RFLP analysis

Bulbus Fritillariae (BF) is the most commonly used antitussive herb in China. There are nine species of Fritillaria recorded as the drug BF in the Chinese Pharmacopoeia. Bulbus Fritillariae cirrhosae (BF cirrhosae) is a group that includes four species of BF; these four species come from wild sources with higher efficiency and lower toxicity compared to the other five species of BF. Due to reasons of carelessness and reduced costs, the other five species are often sold as BF cirrhosae. Analysis through appearance, microscopic and chemical techniques has limitations. Identifying botanical resources is a primary step in the standardization of herbal medicine. In the present article, the internal transcribed spacer 1 (ITS1) regions of the nuclear ribosomal DNA (nrDNA) of nine species and one variety of Fritillaria genus have been sequenced. A mutation site in the ITS1 region among BF cirrhosae and other species of BF has been found and can be recognized by the restriction endonuclease SmaI. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the nuclear ribosomal ITS1 region was used to differentiate BF cirrhosae from other species of BF and is a successful method in distinguishing the subgroups.

Using X-ray diffractometry for identification of Fritillaria preparations according to geographical origin

Fritillaria preparations are traditional herbal medicines used as antitussive and expectorant remedies in China and some other Asian countries. Different Fritillaria species from different geographical origins are difficult to distinguish from each other. We have developed a nondestructive and simple method based on x-ray diffraction to identify five Fritillaria species used for the preparation of medicinals, including F. thunbergii Miq., F. ussurensis Maxim., F. cirrhosa D. Don, F. pallidifloca Schrenk, and F. hupehensis Hsiao et K. C. Hsia. The Fritillaria herbs contain a large amount of starch, which is a semicrystalline macromolecular compound convenient for X-ray diffraction analysis. The difference in the degree of starch crystallinity revealed by X-ray diffraction spectra allowed the Fritillaria species of different geographical origin to be distinguished.

Fritillaria Species Are at Risk of Extinction in Iran: Study on Effective Factors and Necessity of International Attention

The fascinating Fritillaria genus includes 100 species, of which 14 species are native to Iran. Iran is center of origin and genetic diversity of important species like F. imperialis and F. persica. Unfortunately, in recent years wild populations of these valuable plants, which have commercial value, are at risk of extinction in Iran. Effective factors on extinction of F. imperialis and F. persica were investigated in 2 years (2003 and 2004). The important factors were: 1) irregular grazing of Fritillaria stands (provenances); 2) in Iran, there is no protecting rule for saving Fritillaria from extinction; 3) changing of pastures to dry farms; 4) pest overflows that are related to destruction of pastures; and 5) Fritillaria are not commercially cultivated in Iran. Because of poverty, native people eradicate Fritillaria with their bulbs and transfer them to florists' shops. Prevention of this tragedy and conservation of Fritillaria require international attention. Because of these factors, Fritillaria will be removed rapidly from their place of origin.

Inhibitors of Acetylcholine Esterase in vitro - Screening of Steroidal Alkaloids from Fritillaria Species

18 alkaloids were successfully isolated from five Fritillaria species and 5 derivatives were synthesized. Their effects on the bioactivity of human whole blood cholinesterase (ChE) were assessed. The results showed that N-demethylpuqietinone, hupeheninoside, ebeiedinone, yibeinoside A and chuanbeinone inhibited the bioactivity of human whole blood ChE at the concentration of 1.0 x 10 ( - 4) M, with the inhibitory effects of 55.5 +/- 2.7 %, 66.8 +/- 2.0 %, 69.0 +/- 1.7 %, 71.2 +/- 1.8 % and 70.7 +/- 3.3 %, respectively. The effects of the five alkaloids on human red blood cell (RBC) acetylcholinesterase (AChE) and human plasma butyrylcholinesterase (BChE) were further studied, and their IC (50) values for human RBC AChE were 6.4 +/- 0.003 microM, 16.9 +/- 0.018 microM, 5.7 +/- 0.004 microM, 6.5 +/- 0.013 microM and 7.7 +/- 0.001 microM, respectively, and the IC50 values for human plasma BChE were 12.5 +/- 0.026 microM, 2.1 +/- 0.005 microM, 5.2 +/- 0.002 microM, 7.3 +/- 0.005 microM and 0.7 +/- 0.001 microM, respectively. These data suggest, therefore, that N-demethylpuqietinone, hupeheninoside, ebeiedinone, yibeinoside A and chuanbeinone have both anti-RBC AChE and anti-plasma BChE activities, N-demethylpuqietinone is a selective inhibitor of AChE, whereas hupeheninoside and chuanbeinone are the selective inhibitors of BChE.

Identification of the Volatile Component(s) Causing the Characteristic Foxy Odor in Various Cultivars of Fritillaria imperialis L. (Liliaceae)

To identify the component(s) causing the foxy odor, characteristic for some Fritillaria imperialis cultivars, the headspace of flower bulbs was analyzed using gas chromatography-olfactometry (GC-O) and GC-mass spectrometry (GC-MS). Six Fritillaria species and cultivars were selected as follows: F. imperialis cv. Premier (very strong foxy odor), F. imperialis cv. Lutea (strong foxy odor), F. imperialis ssp. Inodora (no odor), Fritillaria eduardii (weak mousy odor), Fritillaria raddeana (no odor), and an F1 of F. imperialis Lutea x Inodora (weak foxy odor). Volatiles from these flower bulbs were accumulated on Tenax and injected into the GC by thermodesorption. The majority of the volatiles consisted of low molecular weight aliphatic compounds. GC-O revealed that the foxy odor was caused by a single component, identified as 3-methyl-2-butene-1-thiol on the basis of smell in GC-O analyses (two GC columns), mass spectra, and retention times. Chemical identification was substantiated by GC-O and GC-MS of an authentic standard of 3-methyl-2-butene-1-thiol, prepared by organic synthesis.

Chemistry, bioactivity and geographical diversity of steroidal alkaloids from the Liliaceae family

Plants belonging to the Liliaceae family have been the topic of research in many phytochemical and pharmacological laboratories because they contain structurally complex and biologically fascinating steroidal alkaloids. This review, citing 153 references, summarises the chemistry, bioactivity and geographical diversity of steroidal alkaloids isolated from Veratrum and Fritillaria species, so as to illustrate the chemo-diversity and biological significance of these alkaloids, along with their geographical distribution where this is discernible.

The Endemic Fritillaria Species of Japan

Summary. This paper reviews and illustrates the seven known species of Fritillaria subgenus Fritillaria that are endemic to Japan and describes a new, eighth species, Fritillaria tokushimensis Akasawa, Katayama et Naito. A key for their identification is provided.

New mesopelagic larvaceans in the genus Fritillaria from Monterey Bay, California

Three new species of Fritillaria are described from specimens collected by a remotely operated vehicle (ROV) between 180 and 735 m depth in Monterey Bay, California. Fritillaria rex becomes the largest species now described for this genus (trunk length up to 10 mm, tail length up to 18 mm). This species is characterized by five or six narrow muscle bundles on each side of the tail, slit-shaped spiracles, a spherical ovary and a cylindrical testis. Fritillaria amphigonadis (trunk length up to 5.2 mm, tail length up to 7.5 mm) is characterized by two narrow muscle bundles on each side of the tail, slit-shaped spiracles, paired spherical ovaries, and a unique pi-shaped testis. Fritillaria lucifer (trunk length up to 4.1 mm, tail length up to 7.0 mm) has 12 extremely thin muscle bundles distributed broadly across the tail, rounded spiracles, a spherical ovary and a U-shaped testis. An additional mesopelagic Fritillaria appears to be consistent with the description of the Mediterranean F. fage, but specific differences in pharyngeal cells, cellular patterns on the tail, and gonadal morphology suggest that the specimens from Monterey Bay are a new variety, F. fagei mbarii. Based on the relatively limited number of collections (i.e. only 20 specimens yielding four unique taxa), it would appear that the possible diversity of larvaceans that live below the surface mixed layer is significantly underestimated.

Crystallography, morphology and thermal properties of starches from four different medicinal plants of Fritillaria species

To fully understand the medicinal plant, Fritillaria, and its species, we investigated the physical properties of starch contained in four Fritillaria species, Fritillaria thunbergii Miq., Fritillaria ussurensis Maxim., Fritillaria pallidifloca Schrenk and Fritillaria cirrhosa D.Don, by means of various analytical methods. The crystal type of the former three kinds of Fritillaria starches was in characteristic B-type, which was in agreement with the crystal type of potato starch. However, the cirrhosa F. starch showed a typical C B-type pattern. The degrees of crystallining of the four Fritillaria starches were about 43.2%, 40.5%, 44.8% and 41.8%, corresponding to thunbergii F. starch, ussurensis F. starch, pallidifloca F. starch and cirrhosa F. starch. The granule sizes of the former two Fritillaria starches ranged from 5 to 40 μm, and were cycloidal or elliptic-shaped. However, the latter two Fritillaria starch granules had granule sizes ranging from 5 to 50 μm, and the granule shape varied from oval to irregular or cuboidal. From the thermogravimetric analysis, it was concluded that the thermal stabilities of the four kinds of starch differed from each other, due to their different structures.

New starches from Fritillaria species medicinal plants

Fritillaria, an important medicinal plant, are widely utilized in the pharmaceutical industry. The bulbs of Fritillaria contain plenty of starches which have been ignored and wasted during the isolation and separation of the small-molecule bioactive ingredients. With the development of food and medicine industry, new starches with different properties have been paid more and more attention. The object of this work is to study the basic physicochemical properties of starch contained in the Fritillaria bulbs. The results revealed that the crystal type of the three Fritillaria starches was the characteristic B-type which was in agreement with the crystal type of potato starch. The results of scanning electron microscope showed that the size of the Fritillaria thunbergii and Fritillaria ussurensis starch granules ranged from 5 to 30 μm, while the Fritillaria pallidifloca starch granules ranged from 5 to 40 μm. The shape of the three Fritillaria starches was also different from each other. The starch separated from F. ussurensis showed the highest enthalpies of gelatinization (Δ H gel) and the lowest transition temperature ( T o, T p, and T c), while the starch from F. thunbergii showed the highest transition temperature, peak height indices (PHI) and the lowest gelatinization temperature range ( R).

BULBOUS MONOCOTS NATIVE TO JAPAN AND ADJACENT AREAS - THEIR HABITATS, LIFE HISTORIES AND PHYLOGENY

The results of comparative studies on the life history characteristics of selected members of bulbous monocots in Japan are reported along with molecular systematic analyses using two chloroplast genes, rbcL and matK. In analyzing the inter- and infra-generic relationships in Lilium and allied taxa, the matK gene was sequenced for 35 Lilium, two Nomocharis, one Cardiocrinum, one Notholirion, and two Fritillaria species, with Erythronium japonicum and Medeola virginiana used as outgroup taxa. Phylogenetic analyses utilizing both MP and NJ topologies of the sequence data demonstrated that the genus Lilium consists of three different clades: the first clade includes 13 Lilium species belonging to Sections Archelirion (5 spp.), Leucolirion (2 spp.), Shinomartagon (5 spp.), Pseudolilium (1 spp.), and two Nomocharis species; the second clade includes 16 Lilium species belonging to Sections Daurolirion (1 sp. with 2 subsp.), Shinomartagon (6 spp.), Martagon (4 spp.), Liriotypus (3 spp.), and Leucolirion (2 spp.); the third clade includes six species belonging to Section Pseudolilium. The two Nomocharis species, N. pardanthina and N. saluenensis, were ingroup taxa of the first clade. Notholirion, Cardiocrinum, and Fritillaria were demonstrated to be sister groups to Lilium. The present results have shown many new evidence concerning the phylogenetic relationships among the Lilium and allied taxa, but do not support Comber’s sectional delimitations (1949).

Identification ofFritillaria pallidifloraUsing Diagnostic PCR and PCR-RFLP Based on Nuclear Ribosomal DNA Internal Transcribed Spacer Sequences

Fritillaria pallidiflora Schrenk (Liliaceae) is a commonly used antitussive herb. There are 9 species of Fritillaria recorded as herbal drugs in the Chinese Pharmacopoeia. The other species are often marketed as F. pallidiflora, and thus, the therapeutic effects of F. pallidiflora are not achieved. Methods to distinguish F. pallidiflora from the 8 other species of Fritillaria are limited by the current morphological and chemical methods. In this study, we report two molecular authentication methods based on the sequences of nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) regions. For diagnostic PCR, we designed a pair of species-specific primers to authenticate F. pallidiflora. The PCR program consisted of only two steps for every repeated cycle. For PCR-RFLP, we identified a distinctive site which can be recognized by the restriction endonuclease Eco81I in the nrDNA ITS1 region of F. pallidiflora. PCR-RFLP analysis was established to differentiate F. pallidiflora from the other species of Fritillaria. These methods provide effective and accurate identification of F. pallidiflora.

New Fritillaria L. taxa from Turkey

A new species of the genus Fritillaria L., F. milasense M. Teksen & Aytac sp. nov. is described from southwest Anatolia in Turkey. It is related to F. bithynica Baker, which mainly differs from it in its leaves, tepal colors, and capsule features. Also, F. serpenticola (Rix) M. Teksen & Aytac stat. nov. is evaluated as a species instead of subspecies of F. carica. Pollen structure and karyotype of F. milasense and F. serpenticola are given.

Determination of the major isosteroidal alkaloids in bulbs of Fritillaria by high-performance liquid chromatography coupled with evaporative light scattering detection

A new direct HPLC analytical method using evaporative light scattering detection coupled with a low-temperature adapter for the simultaneous determination of the major biologically active isosteroidal alkaloids in Bulbus Fritillariae, a commonly used antitussive traditional Chinese medicinal (TCM) herb, has been developed. The simultaneous separation of eight Fritillaria alkaloids was achieved on a reversed-phase C 8 column with an isocratic mobile phase system consisting of acetonitrile–methanol–water (66.5:3.5:30, v/v) containing 0.006% triethylamine. This method provides good reproducibility and sensitivity for the quantification of six major isosteroidal alkaloids, namely peimissine, verticine, verticinone, imperialine, isoverticine and ebeiedine in different Fritillaria species with overall intra- and inter-day precision and accuracy of less than 11% and higher than 90%, respectively. The assay was successfully utilized to quantify the major biologically active alkaloids in five Fritillaria species. The results demonstrate that this method is simple, selective, and suitable for the quality control of this commonly used antitussive TCM herb, Bulbus Fritillariae.

The cultivation of Fritillaria species at Kew

Curtis's Botanical MagazineVolume 17, Issue 3 p. 149-154 The cultivation of Fritillaria species at Kew Richard Wilford, Richard Wilford Senior Botanical Horticulturist in the Alpine and Herbaceous Section at KewSearch for more papers by this author Richard Wilford, Richard Wilford Senior Botanical Horticulturist in the Alpine and Herbaceous Section at KewSearch for more papers by this author First published: 14 January 2004 https://doi.org/10.1111/1467-8748.00259AboutPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Volume17, Issue3August 2000Pages 149-154 RelatedInformation

Simultaneous determination of seven major isosteroidal alkaloids in bulbs of Fritillaria by gas chromatography

The present paper describes the development of a most simple, sensitive, and specific gas chromatographic method to date, for the direct determination of seven major bioactive isosteroidal alkaloids, namely ebeiedine, ebeiedinone, ebeienine, hupehenine, isoverticine, verticine, verticinone and imperialine, in Fritillaria species, a commonly used antitussive traditional Chinese medicinal (TCM) herb. In the present study, a commercially available Supelco SAC-5 capillary column (30 m×0.25 mm, 0.25 μm) specifically designed for the analysis of steroids was utilized for the direct determination of Fritillaria alkaloids. Calibration curves were obtained by spiking authentic compounds and the internal standard (solanidine) into herbal samples prior to extraction. Extraction was conducted simply by shaking the pre-alkalized diethyl ether solution (5.0 ml) containing dried herb (0.1 g) for 2 h. All calibration curves showed good linear regressions ( r 2>0.995) within test ranges. The assay was reproducible and accurate with the overall intra- and inter-day variation and accuracy of less than 10% and more than 90%, respectively. The developed GC method was successfully utilized to analyze seven major bioactive alkaloids in seven Fritillaria species, and the results demonstrate that this direct GC analytical method is suitable for the quality control of this commonly used antitussive TCM herb.