A biometric study of Fritillaria montana Hoppe ex W.D.J. Koch s.l. (Liliaceae) shows a single polymorphic species,with no infraspecific taxa

Abstract

The large amount of morphological variability within Fritillaria montana Hoppe ex W.D.J. Koch, a southeastern European species described from northeastern Italy, led to the creation of several further taxa: Fritillaria caussolensis Goaty & Pons ex Adoino from southeastern France, Fritillaria orsiniana Parl. from central Italy, Fritillaria intermedia N. Terracc. and Fritillaria pollinensis N. Terracc. from southern Italy. Aiming to test the taxonomic value of these taxa, a biometric study of both herbarium and living specimens of F. montana s.l. is carried out. A total of 22 morphological features were analysed and measured in 417 plants from 46 different localities, including loci classici of all the involved taxa. In addition, typical populations of F. montana, F. caussolensis, F. intermedia, F. orsiniana and F. pollinensis and also several other Italian populations resulted with 2n = 18 chromosomes. We also counted 2n = 27 chromosomes in endotriploid cells of plants from Abruzzo. However, there is no clear‐cut correlation between morphology and karyology. The biometric analysis, together with many observations on fresh material in the field, revealed that F. montana is a single polymorphic species with no infraspecific taxa.     

854. FRITILLARIA ARIANA

Fritillaria ariana (Loz.‐Lozinsk. & Vved.) Rix, is illustrated. The history of its discovery and its cultivation are discussed, and a painting and photograph of the habitat are provided.     

Organ culture of a precious Chinese medicinal plant – Fritillaria unibracteata

Fritillaria unibracteata is a famous and precious traditional Chinese medicinal plant. The technique of in vitro organ culture of Fritillaria unibracteata has been established and chemical composition of the metabolites have been confirmed. The experimental results indicated that Fritillaria unibracteata can be rapidly propagated, directly from small cuttings of the bulb by the technique of organ culture. The cultured bulb can be harvested after a 50-day culture period in MS media supplemented with 4.44 μM BA and 5.71 μM IAA. The growth rate was about 30–50 times higher than that under natural wild growth conditions. The content of alkaloid and beneficial micro-elements in the cultured bulbs were higher than found in the wild bulb. It is therefore possible to establish a novel process to produce this natural plant drug in future studies and achieve scale-up for industrial production. This revised version was published online in June 2006 with corrections to the Cover Date.     

Conservation genetics and geographic patterns of genetic variation of the endangered officinal herb Fritillaria pallidiflora

Fritillaria pallidiflora is an endangered officinal herb distributed in the Tianshan Mountains of northwestern China. We examined its phylogeography to study evolutionary processes and suggest implications for conservation. Six haplotypes were detected based on three chloroplast non‐coding spacers (psbA‐trnH, rps16, and trnS‐trnG); genetic variation mainly occurred among populations and SAMOVA groups. This species is distributed in different deep valleys, and we speculate that these fragmented habitats cause gene flow barriers among populations and groups. We also speculate that during glacial periods, extremely low temperatures and aridity caused additional range shrinkage and fragmentation, factors consequently resulting in significant intraspecific differentiation in allopatric regions. For setting a conservation management plan, we identified the Lucaogou region as the most important area, and we designated two ESUs for separate management.     

791. FRITILLARIA SORORUM

Fritillaria sororum J. Persson & K. Persson from the Tauros mountains in southern Turkey is illustrated; its relationship with other species in the area is discussed.     

711. FRITILLARIA HERMONIS

Fritillaria hermonis Fenzl, from Syria and Lebanon is illustrated. Its history and ecology are discussed, and related species are compared. Instructions for its cultivation are given.     

847. FRITILLARIA KIUSIANA

Fritillaria kiusiana L. Hill, a new species from the Kyushu Central Mountains in Japan, is illustrated and described. Its ecology, genetics and conservation are discussed and instructions for its cultivation are given.     

698. FRITILLARIA USURIENSIS

Fritillaria usuriensis Maxim. is illustrated; its cultivation in China for Chinese traditional medicine, and at Kew are described.     

Males are cheaper,or the extreme consequence of size/age‐dependent sex allocation: sexist gender diphasy in Fritillaria montana (Liliaceae)

The bulbous geophyte Fritillaria montana is partially self‐compatible and capable of switching gender. Small flowering plants produce only single male flowers, but larger plants produce hermaphrodite or, rarely, male and hermaphrodite flowers. Eight populations in peninsular Italy were sampled to determine the frequency and size distributions of male and hermaphrodite plants, and to determine the relationship of plant size to male and hermaphrodite flower production. Male plants were significantly smaller than hermaphrodites and made up 14.5–47.8% (100% in one small population) of flowering plants within populations. There were no significant differences in male fitness among female‐sterile and hermaphrodite flowers, as they both possessed full and comparable fertilizing power. Therefore, the gender variation observed in F. montana is likely to depend on resource‐dependent sex allocation. From an evolutionary perspective, we highlight the occurrence of similar mechanisms of gender variation in other representatives of the order Liliales. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 323–333.     

Fritillaria ussuriensis Extract Inhibits the Production of Inflammatory Cytokine and MAPKs in Mast Cells

Fritillaria ussuriensis (FU, derived from the bulbs of various species of the genus Fritillaria, including Fritillaria thunbergii Miq.) is used in herbal medicine to treat conditions such as eczema, skin burns, and frostbite. In this study, we investigated the mechanism of the anti-allergy effect of FU. FU extract (80 mg/kg), orally administered to Sprague-Dawley (SD) rats, significantly inhibited the passive cutaneous anaphylaxis (PCA) reaction. It inhibited the compound 48/80-induced release of histamine from rat peritoneal mast cells in a concentration-dependent manner. Significant inhibitory effects of the FU extract on IL-6, IL-8, and TNF-α (1, 10, and 100 μg/mL) were observed in HMC-1 cells. Treatment with FU attenuated PMA plus A23187-induced phosphorylation of all three MAPKs, especially at concentrations of 10 and 100 μg/mL. Further, it (80 mg/kg) led to significant inhibition of mast-cell accumulation in ear tissue at the chronic phase. These results indicate that it inhibits allergic reactions.     

Phytochemical Variation in <Emphasis Type="Italic">Fritillaria cirrhosa</Emphasis> D. Don (<Emphasis Type="Italic">Chuan Bei Mu</Emphasis>) in Relation to Plant Reproductive Stage and Timing of Harvest<Superscript>1</Superscript>

Phytochemical Variation in Fritillaria cirrhosa D. Don ( Chuan Bei Mu ) in Relation to Plant Reproductive Stage and Timing of Harvest. Economic development in southwest China and the increasing use of traditional Chinese medicine (TCM) worldwide have led to intensified collection of native medicinal plants. Fritillaria cirrhosa D. Don (Chuan Bei Mu), commonly used for the treatment of cough in TCM, is endemic to the Hengduan Mountain region of southwest China and is under increasing pressure from over-collection and decreasing suitable alpine habitat. The bioactive compounds in F. cirrhosa bulbs, isosteroidal alkaloids, are greatly influenced by environmental conditions and fluctuate in content and concentration with plant age and reproductive stage. Aiming at obtaining useful information for the sustainable management of wild F. cirrhosa populations, we evaluated how the phytochemical composition of F. cirrhosa bulbs varies at various stages of plant reproductive development. Using chemical methods and high-performance liquid chromatography, we extracted and analyzed two major bioactive alkaloids from F. cirrhosa bulb samples collected throughout the Hengduan Mountain region. Plant reproductive stage was found to affect the concentration of bioactive alkaloids in F. cirrhosa bulbs. Bulb alkaloid concentration was highest during the early stages of fruit development and decreased significantly with fruit maturation. These results lend biochemical support to the practice of harvesting F. cirrhosa during the early stages of plant senescence (i.e., early fruit development).     

Pathogenic analyses of fungus strains isolated from medicinal Fritillaria przewalskii Maxim. bulb rot

Bulb rot causes a great loss of rare perennial medicinal Fritillaria przewalskii Maxim., which is exacerbated with growth year and seriously constrains the plant productivity, but the pathogens responsible for the disease were still unknown. In this paper, we attempted to explore the potential pathogens leading to bulb rotting and their occurrence patterns in artificial domesticated F. przewalskii. Fungus strains were isolated from the bulb rot spot of the 3‐ to 5‐year‐old diseased F. przewalskii plants by tissue separation, further assayed for pathogenicity according to Koch's law and finally identified by both morphological and molecular methods. Seven pathogenic strains (F1‐F7) were detected in 5‐year‐old rot bulbs, six (F1‐F6) in 4‐year‐old rot bulbs and four (F1, F2, F5, F6) in 3‐year‐old rot bulbs. All of the strains were able to infect bulbs by stabbing and some exhibited varying levels of aggressiveness. Relative to the non‐stabbing controls, the bulbs stab‐inoculated with F5, F2, F7, F4, F1, F6, and F3 showed 76.65%, 75.15%, 71.44%, 40.37%, 39.09%, 36.87%, and 34.93% rot after 8 days, respectively. Phylogenetic analysis revealed that these seven strains were clustered into Bionectria ochroleuca (F1, F3, F4), Fusarium oxysporum (F2, F7), Fusarium tricinctum (F5), and Clonostachys rosea (F6). The two species of Fusarium had the strongest pathogenicity, followed by Bionectria ochroleuca and Clonostachys rosea. Although leading to low bulb rot incidence by stab inoculation, F1 showed the highest isolation rate (48.9%) among all strains. Thus, the edible and medicinal bulbs of F. przewalskii are susceptible to synergetic contamination by these seven pathogens at some point after their third year of growth, which has contributed to the species endangered status, with the two strains of Fusarium being the predominant pathogens. To our knowledge, this is the first report on the seven strains of four fungal species causing F. przewalskii bulb rot in China.     

Appendicularians: an important food supply for the Argentine anchovy Engraulis anchoita in coastal waters

The stomach contents of 532 adult specimens of the Argentine anchovy, Engraulis anchoita, caught in coastal waters of the Argentine Sea from 1994 to 1996, were analyzed. Larger amounts of food were found in the stomachs of anchovies collected in the northern surveys (35–40°S), with the highest values attained in 1994. Main diet items were copepods, appendicularians, cladocerans, fish eggs, and pteropods. Anchovies caught in the southern surveys (40–45°S) ingested less food, whereby the main items were copepods and appendicularians. Plankton samples collected simultaneously with the fishing trawls were also analyzed. Except for Fritillaria borealis, which occurred only in plankton samples, the same appendicularian species (Oikopleura dioica and O. fusiformis) were found in both stomach contents and in plankton samples. The Ivlev selectivity index calculated for O. dioica and O. fusiformis and for the different maturity stages of both species were in all cases ～0, supporting the hypothesis of a non‐selective feeding.     

ABA contents and the regeneration ability of Fritillaria imperialis L. cultured in vitro

Correlation between ABA content in “mother” tissue and subsequent regeneration ability of Fritillaria imperialis cultured in vitro was investigated. In every experiment regeneration was always most efficient from plant material containing the lowest amount of free ABA: a leafy stem part, bulbs used for micropropagation in October and those stored two months at 30 °C prior to in vitro culture. However, no direct correlation between the absolute amounts of ABA and percentage of regeneration was found.     

Variation and taxonomy ofFritillaria graeca (Liliaceae) in Greece

The geographic variation inFritillaria graeca, an endemic of Greece, is explored, using multivariate techniques. Eleven morphological features were measured in each of 630 individual plants from 38 localities throughout the range of the species.Scheffe's a posteriori test revealed an almost continuous geographic variation and did not identify groups clearly. Discriminant analysis of the groups derived from an F significance test ofMahalanobis distance resulted in five major groups on the first two discriminant axes representing the subspeciesgraeca, ionica, guicciardii, thessala and its var.othria. A diagnostic key to theF. graeca subspecies is provided.This paper is a part of a doctoral dissertation dealing withFritillaria in Greece, to be submitted to the department of Biology of the Aristotelian University of Thessaloniki.     

Phylogenetic relationship in Fritillaria spp. of Iran inferred from ribosomal ITS and chloroplast trnL-trnF sequence data

The genus Fritillaria embraces up to 165 taxa in the family Liliaceae, most of which are of high medicinal and ornamental value and importance. In this study, 44 specimens of the genus representing 9 species were collected from their natural habitats located in 10 provinces of Iran. Phylogenetic analysis was performed based on DNA sequences of the internal transcribed spacer (ITS) of the nuclear ribosomal cistron and the trnL-trnF regions. The phylogeny was constructed using the neighbor joining inference method. Results indicate that the examined samples were evidently diverged into 2 distinct clades. Members of the subgenera Fritillaria and Rhinopetalum formed one clade while the other clade contained the subgenera Theresia and Petilium. There can be seen a high degree of similarity between the only yellow-colored crown imperial specimen and the red-colored specimens. The endemic species of Fritillaria straussii, Fritillaria zagrica and Fritillaria kotschyana which their status within the subgenera known in the genus Fritillaria has been remained undefined, fell into the subgenus Fritillaria. The clades also had relatively reasonable distribution patterns based on the genetic structure, geographical conditions and climate specifications. This study revealed the feasibility of the ITS and trnL-trnF DNA sequence for phylogeny of the genus Fritillaria. This is the first phylogenetic analysis of Fritillaria spp. in Iran.     

Inhibition of Sortase,a Bacterial Surface Protein Anchoring Transpeptidase,by β-Sitosterol-3-O-glucopyranoside from Fritillaria verticillata

A glucosylsterol, β-sitosterol-3-O-glucopyranoside, has been isolated as an active principle with sortase inhibitory effect from the bulbs of Fritillaria verticillata by bioassay-guided chromatographic fractionation. The isolate was a potent inhibitor of sortase, with an IC₅₀ value of 18.3 μg/ml and had antibacterial activity against Bacillus subtilis, Staphylococcus aureus, and Micrococcus leuteus with MIC values of 50, 200, and 400 μg/ml, respectively, indicating that this compound is a possible candidate for the development of a bacterial sortase inhibitor. In addition, sitosterol was found to be inactive upon sortase and bacterial cell growth. These results suggest that the inhibitory potency of β-sitosterol-3-O-glucopyranoside is sensitively dependent upon the glucopyranoside side chain moiety.     

Petal: a reliable explant for direct bulblet regeneration of endangered wild populations of <Emphasis Type="Italic">Fritillaria imperialis</Emphasis> L.

Wild populations of Fritillaria imperialis L. are facing extinction and need urgent conservation. This paper presents an efficient system for in vitro direct bulblet regeneration of these populations by petal culturing of flower buds. Petals at different developmental stages, green-closed flower bud (before nectar secretion) and red-closed flower bud (beginning of nectar secretion), were used as explants, and the effects of various proportions of cytokinin to auxin on direct bulblet regeneration pathway were evaluated. More explants switched on direct regeneration pathway in combination of auxins (0.6 mg l⁻¹ NAA + 0.4 mg l⁻¹ IAA) with higher level of cytokinin (1 mg l⁻¹ BAP). In contrast, auxins (0.6 mg l⁻¹ NAA + 0.4 mg l⁻¹ IAA) with lower level of cytokinin (0.1 mg l⁻¹ BAP) produced more bulblets per regenerated explant. In green-closed flower bud stage, direct bulblets regenerated from the end of petal where it was connected to the receptacle, while nectar secretion site was the place of bulblet formation in red-closed flower bud stage. In addition, genotype-dependency of direct bulblet regeneration pathway was investigated by using two different wild populations of Fritillaria imperialis. This plant regeneration procedure was applicable to different Fritillaria genotypes and regenerated bulblets were normal.     

Inhibition of the Bacterial Surface Protein Anchoring Transpeptidase Sortase by Medicinal Plants

Inhibition by medicinal plant extracts of a recombinant sortase was evaluated for antibacterial drug discovery. The coding region of sortase, a transpeptidase that cleaves surface proteins of Gram-positive bacteria, was amplified by PCR from the chromosome of Staphylococcus aureus ATCC 6538p with the exception of an N-terminal membrane anchor sequence, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The purified sortase had maximum activity at pH 7.5 and was stable at 20-45°C for the cleavage of a synthetic fluorophore substrate. The enzyme inhibitory activity in medicinal plants was also evaluated for antibacterial drug discovery. Among 80 medicinal plants tested, Cocculus trilobus, Fritillaria verticillata, Liriope platyphylla, and Rhus verniciflua had strong inhibitory activity. The extract with the greatest activity was the ethyl acetate fraction derived from the rhizome of Cocculus trilobus (IC₅₀=1.52 μg/ml).