Lyconadins D and E, and complanadine E, new Lycopodium alkaloids from Lycopodium complanatum

Three new Lycopodium alkaloids, lyconadins D (1) and E (2), and complanadine E (3), were isolated from the club moss Lycopodium complanatum. Lyconadin D (1) was the first example of fastigiatine-type alkaloid isolated from Lycopodium complanatum. The structures and relative stereochemistry of 1-3 were elucidated on the basis of spectroscopic data. Complanadine E (3) enhanced mRNA expression for NGF.     

Lycoparins A-C, new alkaloids from Lycopodium casuarinoides inhibiting acetylcholinesterase

Three new Lycopodium alkaloids, lycoparins A-C (1-3), have been isolated from the club moss Lycopodium casuarinoides. Structures and stereochemistry of 1-3 were elucidated on the basis of 2D NMR correlations. Lycoparins C (3) exhibited an inhibitory activity against acetylcholinesterase, while lycoparins A (1) and B (2) did not show activity.     

Interactions of Lycopodium alkaloids with acetylcholinesterase investigated by 1H NMR relaxation rate

In order to further understand the interaction processes between the Lycopodium alkaloids and acetylcholinesterase, the binding properties of N-acetyl huperzine A (1), huperzine B (2) and huperzine F (3) with Torpediniforms Nacline acetylcholinesterase (TnAchE) were investigated by 1H NMR methods. The nonselective, selective and double-selective spin-lattice relaxation rates were acquired in the absence and presence of TnAchE at a ratio of [ligand]/[protein]=1:0.005. The selective relaxation rates show protons of 1-3 have dipole-dipole interaction with protons of TnAchE at the binding interface. The molecular rotational correlation time of bound ligands was calculated by double-selective relaxation rate at 298 K, which showed that 1-3 had high affinity with the protein. The results indicate that investigation of 1H NMR relaxation data is a useful method to locate the new Lycopodium alkaloids as AchE inhibitors.     

Complanadines C and D, new dimeric alkaloids from Lycopodium complanatum

Two new dimeric Lycopodium alkaloids, complanadines C (1) and D (2), have been isolated from the club moss Lycopodium complanatum, and the structures and relative stereochemistry of 1 and 2 were elucidated on the basis of the spectral data. Complanadine D (2) enhanced mRNA expression for NGF.     

Lycopladines B-D and lyconadin B, new alkaloids from Lycopodium complanatum

Four new alkaloids, lycopladines B-D (1-3) and lyconadin B (4), have been isolated from the club moss Lycopodium complanatum and the structures including the stereochemistry were elucidated on the basis of spectral data and modified Mosher's method. Lyconadin B (4) elevated NGF mRNA expression in 1321N1 human astrocytoma cells.     

东北石杉生物碱成分的研究

从东北石杉中分离得到7个石松生物碱,通过NMR、MS及IR等光谱分析,分别鉴定为：luciduline(1),dithydroluciduline(2),lycocline(3),huperzine B(4),serratinine(5),serratidine(6),mecleanine(7)。这些化合物均为首次从该植物中分得。     

马尾伸筋草化学成分及抗骨质疏松活性研究

为研究马尾伸筋草(Lycopodium fargesii Hert.)的化学成分及其抗骨质疏松活性,采用硅胶、MCI、RP-18、Sephadex LH-20柱色谱及半制备高效液相色谱对马尾伸筋草的乙醇提取物进行系统分离,从乙酸乙酯部位中得到18个化合物,综合利用NMR、HR-ESI-MS等技术,分析鉴定化合物的结构,分别鉴定为α-芒柄蜡素(1)、26-去甲-8-氧化-α-芒柄蜡素(2)、serrat-14-en-3β,21α,24-triol(3)、serrat-14-en-3β,21β,24-triol(4)、1,20-eicosanediol(5)、二十三烷(6)、glycerolmonolinoleate(7)、glyceryl linolentate(8)、3-(4′-formylphenoxy)-4-methoxybenzaldehyde(9)、芹菜素(10)、对羟基苯甲醛(11)、香草酸(12)、3-羟基-4-甲氧基苯甲酸(13)、2-羟基-1-(4-羟基-3-甲氧基-苯基)-丙基-1-酮(14)、3-甲氧基-4-羟基桂皮醛(15)、8-acetoxy-5-hydroxyumbelliprenin(16)、trans-4-hydroxy-2-nonenoic acid(17)和正十烷硫醇(18)。化合物1~18均为首次从马尾伸筋草中分离得到,其中化合物16对破骨细胞活性具有显著的抑制作用,其IC₅₀为1.98μM。     

Molecular cloning and characterization of copper amine oxidase from Huperzia serrata

A cDNA encoding a novel copper amine oxidase (CAO) was cloned and sequenced from the Chinese club moss Huperzia serrata (Huperziaceae), which produces the Lycopodium alkaloid huperzine A. A 2043-bp open reading frame encoded an Mr 76,854 protein with 681 amino acids. The deduced amino acid sequence shared 44-56% identity with the known CAOs of plant origin, and contained the active site consensus sequence of Asn-Tyr-Asp/Glu. The phylogenetic tree analysis revealed that HsCAO from the primitive vascular plant H. serrata is closely related to Physcomitrella patens subsp CAO. The recombinant enzyme, heterologously expressed in Escherichia coli, catalyzed the oxidative deamination of aliphatic and aromatic amines. Among them, the enzyme accepted cadaverine as the best substrate to catalyze the oxidative deamination to Δ(1)-piperideine, which is the precursor of the Lycopodium alkaloids. Furthermore, a homology modeling and site-directed mutagenesis studies predicted the active site architecture, which suggested the crucial active site residues for the observed substrate preference. This is the first report of the cloning and characterization of a CAO enzyme from the primitive Lycopodium plant.     

Characterization of the Selaginella remotifolia MADS-box gene

Recent progress in plant molecular genetics has revealed that floral organ development is regulated by several homeotic selector genes, most of which belong to the MADS-box gene family. Here we report on SrMADS1,a MIKC(c)-type MADS-box gene from Selaginella, a spikemoss belonging to the lycophytes. SrMADS1 phylogenetically forms a monophyletic clade with genes of the LAMB2 group, which are MIKC(c) genes of the clubmoss Lycopodium, and is expressed in whole sporophytic tissues except roots and rhizophores. Our results and the previous report on Lycopodium MIKC(c) genes suggest that the ancestral MIKC(c )gene of primitive dichotomous plants in the early Devonian was involved in the development of basic sporophytic tissues such as shoot, stem, and sporangium.     

Arbuscular mycorrhizal associations in Lycopodiaceae

This study characterizes the molecular and phylogenetic identity of fungi involved in arbuscular mycorrhizal (AM) associations in extant Huperzia and Lycopodium (Lycopodiaceae). Huperzia and Lycopodium are characterized by a life cycle with long-lived autotrophic sporophytes and long-lived mycoheterotrophic (obtain all organic carbon from fungal symbionts) gametophytes. 18S ribosomal DNA was isolated and sequenced from Glomus symbionts in autotrophic sporophytes of seven species of Huperzia and Lycopodium and mycoheterotrophic Huperzia gametophytes collected from the Páramos of Ecuador. Phylogenetic analyses recovered four Glomus A phylotypes in a single clade (MH3) that form AM associations with Huperzia and Lycopodium. In addition, phylogenetic analyses of Glomus symbionts from other nonphotosynthetic plants demonstrate that most AM fungi that form mycoheterotrophic associations belong to at least four specific clades of Glomus A. These results suggest that most mycoheterotrophic plants that form AM associations do so with restricted clades of Glomus A. Moreover, the correspondence of identity of AM symbionts in Huperzia sporophytes and gametophytes raises the possibility that photosynthetic sporophytes are a source of carbon to conspecific mycoheterotrophic gametophytes via shared fungal networks.     

Karyomorphology of the genusLycopodium sensu stricto and relationships among species

Karyomorphological comparisons were made of six species of JapaneseLycopodium sensu stricto. There were no marked differences at interphase and prophase among the six species.Lycopodium annotinum had 2n=68 and the formula of its metaphase karyotype was 18m(median centromeric chromosomes)+12sm(submedian)+12st(subterminal)+26t(terminal).Lycopodium casuarinoides had 2n=68=16m+10sm+18st+24t,L. clavatum 2n=68=22m+12sm+18st+16t, andL. obscurum 2n=68=10m+22sm+20st+16t. Each of these species, which belong to different sections, displayed several karyomorphological differences. Among themL. casuarinoides differs largely from the others in its mean chromosome length, ratio of the longest chromosome to the shortest, and frequency of m+sm chromosomes. BothLycopodium complanatum andL. nikoense, belonging to sectionComplanata, had a common karyotype 2n=46=10m+12sm+18st+6t. This section displayed a low differentiation in its karyotype. In the wholeLycopodium s.s., the ratios of m+sm in a complement varied from 38 to 50%, being higher among pteridophytes.     

伸筋草类药用植物资源的初步研究

本文报道6种商品伸筋草类药用植物的本草考证、理化分析及药理试验。豚鼠离体肠平滑肌实验结果表明:玉柏石松(Lycopodium obscurum L.)和华中石松(L.centro-chinese Ching)的总提取物有较强兴奋作用,其次是扁枝石松(Diphasiastrum complanatum (L.) Holub),再次是垂穗石松(Palhimhaea cernua (L.) Franco et Vase)、石松(Lycopodium japonicum Thunb.)及藤石松(Lycopodiastrum casuarinodes (Spring) Holub)。这与各自的总生物碱含量相吻合。从抗血小板凝聚试验来看。玉柏石松总提取物作用最强,其次是扁枝石松。     

GAMETOPHYTE OF LYCOPODIUM DIGITATUM

Gametophytes of Lycopodium digitatum A. Braun are subterranean, mycorrhizal structures. Externally, they are conical and divided into three principal regions: tapering base, ring meristem, and gametangial cap. Internally, the endobiont is restricted to specialized tissues in the base. Gametangia typical of those described for L. complanatum are found along the cap margin near the ring meristem. Comparative analysis of these and other related gametophytes leads to the conclusion that Bruchmann's Type I (L. clavatum) and Type II (L. complanatum) gametophytes cannot be consistently separated on the basis of 1) mycorrhizal tissues, 2) form of gametophyte, and 3) number of neck canal cells in the archegonium, as he had proposed. Consequently, it is suggested on the basis of ring meristem and radial symmetry, mycorrhizal tissue specialization, and lack of sterile paraphyses or lobes that Type I and Type II gametophytes be merged as a single type. The now broadened clavatumcomplanatum type is restricted to subgenus Lycopodium (sensu Wilce) where it is the only known type.     

Validation of generic names inLycopodiaceae: with a description of a new genusPseudolycopodiella

Four earlier proposed and invalidly published generic names ofLycopodiaceae are validated:Lateristachys (type:Lycopodium laterale R. Br.),Lycopodiastrum (type:Lycopodium casuarinoides Spring),Pseudodiphasium (type:Lycopodium volubile G. Forster) andPseudolycopodium (type:Pseudolycopodium densum (Rothm.) Holub). A new genusPseudolycopodiella (type:Lycopodium carolinianum L.) is described. New nomenclatural combinations are proposed for 17 species of these genera and ofLycopodiella.     

Diphaladine A，扁枝石松中一个新的石松生物碱

从扁枝石松（Diphasiastrum complanatum）的95％乙醇提取物中分离得到1个新的石松生物碱,经波谱技术鉴定其结构,命名为diphaladine A（1）。同时还分离得到其它9个已知石松生物碱类化合物,其中obsctmmaine A（2）,L20,lycoposerramine—K,des—N—methyl—β—obscurine,des-N—methyl-α—obscurine,lycoflexion,和phlegmariurine B等7个化合物均为首次从该植物中分离得到。     

Capsules with Lycopodium spores for absolute diatom and pollen analysis

Lycopodium spore capsules have been produced as a slight modification of the widely used Lycopodium spore tablets introduced by Stockmarr in 1971. The spore content is increased in the capsules to facilitate their use in absolute diatom analysis.     

Serratezomines D and E,new Lycopodium alkaloids from Lycopodium serratum var. serratum

Two new Lycopodium alkaloids, serratezomines D (1) and E (2), were isolated from the club moss Lycopodium serratum var. serratum. Serratezomine D (1) is a new lucidine-type alkaloid, while serratezomine E (2) is a new phlegmarane-type alkaloid. The structures and relative stereochemistry of 1 and 2 were elucidated on the basis of spectroscopic data. Serratezomine D (1) exhibited an inhibitory activity against acetylcholinesterase.     

香港蕨类植物新记录

报道了香港蕨类植物新记录种5种,即台湾轴脉蕨(Ctenitopsis kusukuensis)、掌叶线蕨(Colysis digi-tata)、德化鳞毛蕨(Dryopteris dehuaensis)、海岛鳞始蕨(Lindsaeaorbiculatavar.commixta)和石松(Lycopo-diumclavatum),其中包括两个新记录属轴脉蕨属(Ctenitopsis)和石松属(Lycopodium)。另外,还列出了每个种的重要特征及其与相关种类的差异。     

Vascular architecture in shoots of early divergent vascular plants, Lycopodium clavatum and Lycopodium annotinum

Lycopodium represents a phylogenetically distinct clade of basal vascular plants with anatomical characters that have no parallel in other lineages. Thus, knowledge of lycopod structure and development may reveal important information about the common ancestors of all vascular plants. Here we report the unique architecture of the conducting system in Lycopodium annotinum and Lycopodium clavatum. Based on multiple series of anatomical sections, we reconstructed spatial relationships between microphylls and the stelar system. Analysis revealed that protoxylem ribs (PXR) were vertical, regardless of type of phyllotaxis, and their numbers were variable. Microphyll traces (MTr) were randomly distributed between ribs, resulting in the absence of defined sympodia and varied lengths of MTr. Dichotomous branching contributed to additional features, for example occurrence of mesarch protoxylem, affecting stele structure and PXR numbers. Our data showed limited interrelationships between lycopod vasculature and microphyll phyllotaxis. This may suggest that both systems developed independently, then evolved together to form the integrated supply system. Thus vasculature in extant lycophytes may be less functionally efficient than in seed plants, where consistent leaf-trace lengths guarantee predictable energy utilization during ontogeny. Differences may result from the phylogenetically different origin of microphylls, and the level of vascular complexity.     

3-O-methyl-D-galactose residues in lycophyte primary cell walls

Acid hydrolysis of cell wall-rich material from young leaves of the lycophyte Selaginella apoda (L.) Spring yielded substantial amounts of 3-O-methyl-D-galactose (1) in addition to the usual major monosaccharides (glucose, galactose, arabinose, xylose and galacturonic acid). The yield of 1 approximately equalled that of galacturonic acid. Compound 1 was identified as 3-O-methylgalactose by its 1H and 13C NMR spectra, and shown to be the D-enantiomer by its susceptibility to D-galactose oxidase. Compound 1 was detected in acid hydrolysates of the alcohol-insoluble residues from young leaves of all lycophytes tested, both homosporous (Lycopodium, Huperzia and Diphasiastrum) and heterosporous (Selaginella). It was not detectable in the charophyte green algae Coleochaete scutata, Chara coralina or Klebsormidium flaccidum, any bryophytes [a hornwort (Anthoceros), four liverworts and three mosses], or any euphyllophytes [a psilopsid (Psilotum), a horsetail (Equisetum), eusporangiate and leptosporangiate ferns, the gymnosperm Gnetum, and diverse angiosperms]. A high content of 1 is thus an autapomorphy of the lycophytes.