The Influence of Aerial Exposure on Sea Anemones Aulactinia veratra Mucin Genes Expression Using the RNA Sequencing

Mucin genes are the main component of mucus. The sea anemone species, Aulactinia veratra (Phylum Cnidaria) contains different types of mucin genes. In the intertidal zone, A. veratra is found to be exposed to air during the low tide and produces large quantities of mucus as an external covering. The relation between low tide and mucus secretion is still unclear, and what is the role of mucin during arial exposure is not yet investigated. This study hypothesised that the mucin genes in A. veratra would have significantly high expression in response to aerial exposure. Therefore, the aim of current study was to examine and analyses the response of A. veratra mucins in response to an experiment involving three hours of aerial exposure. To achieve this, aim the RNA-sequencing and bioinformatics analyses were used to examine the expression profile of A. veratra mucin genes in response to aerial exposure. The generated results have shown that, Mucin4-like and mucin5B-like were up-regulated in response to the three hours of aerial exposure in A. veratra. This finding shows a significant role of mucin5B-like and mucin4-like genes in response to air stress at low tide. The data generated from this study could be used in conjunction with future mucin gene studies of sea anemones and other cnidarians to compare A. veratra mucin gene expression results across time, and to extend our understanding of mucin stress response in this phylum.

Isolation and identifi cation of antimicrobial metabolites from sea anemone-derived fungus Emericella sp.SMA01

Marine symbiotic fungi represent an intriguing source of discovery of novel secondary metabolites with various biological activities.Sea anemones are benthic marine invertebrates,however,the cultivable symbiotic fungi residing in the sea anemones are paid few attentions compared to those derived from their cnidarian counterparts.Here we show the identification of antimicrobial secondary metabolites from the sea anemone-derived symbiotic fungi.Out offive isolated fungal strains,only the strain SMA01 showed strong antimicrobial activities,which was assigned into the genus Emericella based on the morphological characteristics and the ITS sequencing.Media swift from liquid fermentation to solid rice medium presented little influence on its antibacterial activity.A chemical investigation of the ethyl acetate extract of the Emericella sp.SMA01 led to discovery of the primary antibiotic metabolite phenazine-1-carboxylic acid.The IC_(50) values of the phenazine-1-carboxylic acid against Phytophthora capsici,Gibberella zeae,and Verticillium dahliae were determined to be 23.26-53.89μg/mL.To the best of our knowledge,this was the first report of Emericella sp.in sea anemones.The current study may benefit understanding of the defensive chemical interactions between the symbiotic fungi and their host sea anemones.

Antitumor triterpene saponins from Anemone flaccida

A new triterpenoid saponin, 3-O-[（6′-butyryl）-β-D-glucopyranosyl]-28-O-[α-L-rhamnopyranosyl-（1→4）-β-D-glucopyranosyl-（1→6）-β-D-glucopyranosyl] oleanolic acid, as well as three known triterpenoid saponins were isolated from the rhizomes of Anemone flaccida. Their structures were elucidated by spectroscopic methods. These compounds showed significant antitumor activities.

A New β-Carboline Alkaloid and a New Derivate of Isoferulic Acid from Anemone altaica

A new β-carboline alkaloid, 4-（9H-β-carbolin-1-yl）-4-oxobutyric acid and a new derivate of isoferulic acid, （E）-3-（3-hydroxy-4-methoxyphenyl）acrylic acid carboxymethyl ester, were isolated from the roots of Anemone altaica. Their structures were determined on the basis of spectral data.

Two New Oleanane Saponins from Anemone anhuiensis

Two new oleanane triterpene saponins, named as anhuienside C (1) and D (2), were isolated from the rhizomes of Anemone anhuiensis (Ranunculaceae). The structures of the new compounds were elucidated as 3-O-alpha-L-rhamnopyranosyl (1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl ester (1) and 3-O-beta-D-xylopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl ester (2), respectively.

Antimicrobial properties of sea anemone Anthopleura nigrescens from Pacific coast of Costa Rica

Objective: To evaluate the antimicrobial and antifungal activities of the aqueous and partitioned extract of sea anemone Anthopleura nigrescens(A. nigrescens).Methods: The sea anemone A. nigrescens was collected, minced, homogenized,lyophilized and then further partitioned with diethyl ether, acetone, ethanol and water.These fractions were evaluated for antimicrobial activity against bacterial and fungal pathogens.Results: Acetone extract was found to produce a pronounced inhibition of 7.0 mm against Proteus vulgaris and diethyl ether extract inhibited Pseudomonas aeruginosa with an inhibition zone of 6.5 mm. In antifungal activity, ethanol extract showed good activity against Botrytis cinerea, Trichoderma harzianum and Rhizopus oryzae compared with other strains. Acetone and ethanol extract of A. nigrescens showed activity against all of pathogens tested. Slight activity was observed in the water extract with inhibition zone of 1.5 mm.Conclusions: The present study revealed that sea anemone A. nigrescens may also contain some biologically active agents which have potential activity against pathogenic microorganisms.

A New Triterpenoid Saponin from Anemone tomentosa

A new triterpenoid saponin, tomentoside (5), together with prosapogenin CP 4 (1),huzhangoside B (2), C (3) and D (4) was isolated from the ethanolic extract of Anemone tomentosa.its structure was established by spectroscopic and chemical evidence.

A new triterpenoid saponin from Anemone raddeana

A new triterpenoid bisglycoside, named raddeanoside R19, was isolated from the roots of Anemone raddeana. Its structure was elucidated on the basis of ESI-MS, ^13C NMR, and chemical methods.

A new superoxide-generation inhibitor from rhizome of Anemone raddeana Regel

A new triterpenoid saponins, raddeanoside R19 （1） was isolated from the rhizome of Anemone raddeana Regel. The effects of raddeanoside R119 on superoxide generation in human neutrophil were evaluated. The compound suppressed the superoxide generation induced by N-formyl-methionyl-leucyl-phenylalanine （fMLP）, phorbol 12-myristate 13-acetate （PMA） and arachidonic acid （AA） in a different concentration-dependent manner.

Anemhupehins A-C,Podocarpane Diterpenoids from Anemone hupehensis

Three new podocarpane diterpenoids,namely anemhupehins A-C(1-3),together with four known analogues(4-7),have been isolated from aerial parts of Anemone hupehensis.Their structures were characterized based on extensive spectroscopic data.Compounds 1 and 4 showed certain cytotoxicities against human cancer cell lines.

Preparation and Insecticidal Activity of Sea Anemone Peptide AP-GI from Aiptasia pallida

[Objectives] This study was conducted to synthesize sea anemone peptide Ap-GI and investigate its insecticidal activity. [Methods] The linear peptide Ap-GI was synthesized by solid phase peptide synthesis(SPPS), and the linear peptide was subjected to the two-step oxidative folding, mass spectrometry identification and high performance liquid chromatography(HPLC) purification. Then, the MTT method and insect injection method were used to study its insecticidal activity. [Results] The synthesized sea anemone peptide had a purity of 95%. The test results of the MTT method showed that the peptide Ap-GI had the activity of inhibiting the growth of insect cells sf9 with the median effective dose of 0.7 nM;and the test results of the injection method on yellow mealworms showed that the peptide Ap-GI had high insecticidal activity, and the median lethal dose was 16.9 nM. [Conclusions] The sea anemone peptide Ap-GI from Aiptasia pallida has a good inhibitory effect on the growth of insect cells and high-efficiency insecticidal activity, which can lay a foundation for the development of new, safe and efficient peptide biological insecticides.

Preparation of Sea Anemone Peptide Toxin Ap-TxI and Its Insecticidal Activity

[Objectives]This study was conducted to synthesize sea anemone peptide toxin Ap-TxI and investigate its insecticidal activity. [Methods] The sea anemone linear peptide toxin Ap-TxI was synthesized by the solid-phase peptide synthesis(SPPS), and six cysteines were oxidized to form three disulfide bonds by a three-step directional oxidation method. Then, purification by high performance liquid chromatography(HPLC) and mass spectrometry identification were performed. Finally, the insect cytotoxicity and insecticidal activity of Ap-TxI were studied by the MTT method and insect injection method. [Results] The oxidized peptide Ap-TxI with three disulfide bonds in natural configuration was successfully synthesized by the SPPS method, and its purity was >90% by HPLC analysis. The results of the MTT method showed that Ap-TxI was active on the growth of insect cells sf9, with a half effective dose of 0.2 nM;and the results of the mealworm injection test showed that the polypeptide Ap-TxI had high insecticidal activity with a median lethal dose of 11.7 nM. [Conclusions] The sea anemone peptide toxin Ap-TxI with high insecticidal effect was obtained, laying a foundation for the development of new, efficient and safe biological insecticides.

Transcriptomic analysis in Anemone flaccida rhizomes reveals ancillary pathway for triterpene saponins biosynthesis and differential responsiveness to phytohormones

Anemone flaccida Fr. Schmidt is a perennial medicinal herb that contains pentacyclic triterpenoid saponins as the major bioactive constituents. In China, the rhizomes are used as treatments for a variety of ailments including arthritis. However, yields of the saponins are low, and little is known about the plant's genetic background or phytohormonal responsiveness. Using one-quarter of the 454 pyrosequencing information from the Roche GS FLX Titanium platform, we performed a transcriptomic analysis to identify 157 genes putatively encoding 26 enzymes involved in the synthesis of the bioactive compounds. It was revealed that there are two biosynthetic pathways of triterpene saponins in A. flaccida. One pathway depends on β-amyrin synthase and is similar to that found in other plants. The second, subsidiary("backburner") pathway is catalyzed by camelliol C synthase and yields β-amyrin as minor byproduct. Both pathways used cytochrome P450-dependent monooxygenases(CYPs) and family 1 uridine diphosphate glycosyltransferases(UGTs) to modify the triterpenoid backbone. The expression of CYPs and UGTs were quite different in roots treated with the phytohormones methyl jasmonate, salicylic acid and indole-3-acetic acid. This study provides the first large-scale transcriptional dataset for the biosynthetic pathways of triterpene saponins and their phytohormonal responsiveness in the genus Anemone.

Anemone medicinal plants:ethnopharmacology,phytochemistry and biology

The Ranunculaceae genus Anemone(order Ranunculales),comprising more than 150 species,mostly herbs,has long been used in folk medicine and worldwide ethnomedicine.Various medicinal compounds have been found in Anemone plants,especially triterpenoid saponins,some of which have shown anti-cancer activities.Some Anemone compounds and extracts display immunomodulatory,antiinflammatory,antioxidant,and antimicrobial activities.More than 50 species have ethnopharmacological uses,which provide clues for modern drug discovery.Anemone compounds exert anticancer and other bioactivities via multiple pathways.However,a comprehensive review of the Anemone medicinal resources is lacking.We here summarize the ethnomedical knowledge and recent progress on the chemical and pharmacological diversity of Anemone medicinal plants,as well as the emerging molecular mechanisms and functions of these medicinal compounds.The phylogenetic relationships of Anemone species were reconstructed based on nuclear ITS and chloroplast markers.The molecular phylogeny is largely congruent with the morphology-based classification.Commonly used medicinal herbs are distributed in each subgenus and section,and chemical and biological studies of more unexplored taxa are warranted.Gene expression profiling and relevant "omics" platforms could reveal differential effects of phytometabolites.Genomics,transcriptomics,proteomics,and metabolomics should be highlighted in deciphering novel therapeutic mechanisms and utilities of Anemone phytometabolites.

Floral Morphogenesis of Anemone rivularis Buch.-Ham. ex DC. var.flore-minore Maxim. （Ranunculaceae） with Special Emphasis on Androecium Developmental Sequence

: The floral morphogenesis and androecium developmental sequence of Anemone rivularis Buch.-Ham. ex DC. var. flore-minore Maxim. were observed under a scanning electron microscope (SEM) and by means of histological methods in order to expand our knowledge of the morphogenesis and development of the floral organs of the Ranunculaceae. The initiation of the floral elements is a centripetal spiral and the direction of the spiral is clockwise or anti-clockwise. However, the development of the androecium is highly unusual: in a longitudinal series of four stamens, the second stamen develops first from the inner to outer, then the third one, the fourth one and the first one in turn. The microsporogenesis and anther maturation follows the same developmental sequence. The tepals are different from the bracts and the stamens in both shape and size in the early developmental stage, but there is no difference between the stamens and carpels in the early developmental stage. Therefore, we established a spatio-temporal process of the floral morphogenesis of A. rivularis var. flore-minore and offer another meaning of the floral diversity patterns attributed to the level of the genus.

STUDIES ON THE CHEMICAL CONSTITUENTS OF THE CHINESE MEDICINAL HERB ANEMONE RADDEANA REGEL

The Chinese medicinal herb, Anemone raddeana Regel, has been used for treatment of rheumatism, phlebitis, etc. in folk medicine. Saponins R4, R5, R6, R7 have been isolated from this plant.

Chemical Constituents of the Roots of Anemone altaica Fisch. ex C. A. Mey.

The roots of Anemone altaica Fisch. ex C. A. Mey. have been used in the treatment of epilepsia, neurasthenia, and arthritis in Chinese folk medicine for a long time. In order to find new and bioactive compounds, the chemical constituents of the roots of A. altaica were investigated and nine compounds were isolated from the EtOH extract of this plant. On the basis of spectroscopic methods, the structures of these compounds were elucidated as 4-（9H-β-carbolin- 1-yl）-4-oxo-butyric acid （1）, carboxymethyl isoferulate （2）, isoferulic acid （3）, cirsiumaldehyde （4）, 5-hydroxy-4-oxo-pentanoic acid （5）, triacontane （6）, palmic acid （7）, β-sitosterol （8）, and daucosterol （9）. Among them, 1 and 2 were new compounds, and 3 and 4 were obtained from this genus for the first time.

New Triterpenoid Saponins from the Rhizomes of Anemone amurensis

Five new oleanane-type saponins （1-5）, together with three known ones （6-8）, were isolated from the rhi- zomes of Anemone amurensis. Their structures were determined on the basis of spectroscopic analysis and chemical evidence. Compounds 2-4 are three novel sulfated oleanane-type triterpenoid glycosides.

Review of Anemone Raddeana Rhizome and Its Pharmacological Effects

The chemical compositions of Anemone raddeana Rhizome, a kind of traditional Chinese medicine, were reviewed, along with its bioactivity and pharmacological properties and method improvements of extracting and detecting triterpenoid saponins. A. raddeana Rhizome is used to treat neuralgia and rheumatism, and is rich in triterpenoid saponins, most of which are pentacyclic, with oleanane as the nucleus. So far, 37 triterpenoid saponins have been determined from the herb. Its reported bioactivity and pharmacological properties have been described as anticancerous, antimicrobial, anti-inflammatory, analgesic, antipyretic, anticonvulsive, antihistaminic, and sedative. It has also been used for the induction of the humoral immune response and treatment of liver fibrosis in chronic hepatitis. However, the herb also has hemolytic effects and can be toxic, which limits its clinical application. Further studies are needed on the pharmaceutical functions, mechanisms, and immunological responses to contribute to the herb＇s clinical applications.

Effects of Crude Venom From Sea Anemone Authopleura qingdaoenisis on Neuromuscular Preparation: Its Mode and Sites of Action

1 Introduction Sea anemone toxins are polypeptide neurotoxins. The symptoms of the toxic animals exhibits contracture and paralysis of their limbs. It is evident that the peripheral nerve and muscle are important targets of the toxin. Up to now, it is still a controversial topic about which part of the N-M preparation is affected. Shapiro considered that the venom from Condylactic gigantea was active on crayfish nerve but not on muscle membrane. Metezeau et al. used curarized and denervated frog sartofius muscles, and found that ATX-Ⅱ from Anemonia sulcata acted on motor nerve terminals but not directly on muscle fibres. Erxleben et al. proved that ATX-Ⅱ could act on frog skeletal muscle directly