OBJECTIVETo establish and optimize the cultivation systems for wild Huperzia serrata, through assessing the effects of environmental factors on its growth.

METHODH. serrata was planted under natural field condition, the growth status of H. serrata under different light intensities, temperatures, relative humidity (R. H.), fertilizer levels, and so on were investigated and analyzed.

RESULTH. serrata grew nicely under following environmental conditions: temperature ranging from 18.9-26.3 degrees C, R. H. ranging from 81%-90% and 1330-3000 lx light intensity. A cultivation system, in which H. serrata was planted in the bamboo forest and did not occupy the arable land, was established through the study.

CONCLUSIONConcluded that light intensity and R. H are the two factors which greatly confine the growth of H. serrata. As long as the levels of the above two factors meets the demands, H. serrata could be cultivated.

Fertilizers ; Humidity ; Huperzia ; growth & development ; Light ; Temperature

A total of 127 strains of endophytic fungi were isolated from roots, branches and leaves of Huperzia serrata. These strains were identified into 19 genera based on morphological characters and ribosomal DNA (rDNA) sequence analysis, there into Penicillium, Aspergillus and Podospora were dominant populations in H. serrata. From analysis results we found some endophytic fungi showed a certain degree of tissue preference. The isolation rate and colonization rate of stems were both larger than those of leaf and roots. After testing the acetylcholinesterase (AChE) inhibitory activity of these endophytic fungi, a total of 39 endophytic fungi belonging to 15 genera showed AChE inhibition. Eleven endophytic fungi showed potent AChE inhibition, 7 of which were isolated from leaf. The research not only provided theoretical basis for developing and utilizing the resources of endophytic fungi in H. serrata but also showed a new path for searching medicines resource which has AChE inhibitory activity.
Cholinesterase Inhibitors ; pharmacology ; Fungi ; classification ; isolation & purification ; Huperzia ; microbiology

Xanthocillin is a unique natural product with an isonitrile group and shows remarkable antibacterial activity. In this study, the genome of an endophytic fungus Penicillium chrysogenum MT-40 isolated from Huperzia serrata was sequenced, and the gene clusters with the potential to synthesize xanthocillin analogues were mined by local BLAST and various bioinformatics analysis tools. As a result, a biosynthetic gene cluster (named for) responsible for the biosynthesis of xanthocillin analogues was identified by further heterologous expression of the key genes in Aspergillus oryzae NSAR1. Specifically, the ForB catalyzes the synthesis of 2-formamido-3-(4-hydroxyphenyl) acrylic acid, and the ForG catalyzes the dimerization of 2-formamido-3-(4-hydroxyphenyl) acrylic acid to produce the xanthocillin analogue N, N'-(1, 4-bis (4-hydroxyphenyl) buta-1, 3-diene-2, 3-diyl) diformamide. The results reported here provide a reference for further discovery of xanthocillin analogues from fungi.
Penicillium chrysogenum/genetics* ; Huperzia/microbiology* ; Acrylates ; Multigene Family

AIMTo study the alkaloid constituents of Huperzia serrata (Thunb.) Trev..

METHODSChromatographic methods were used for the isolation and purification. Structure was elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSAn alkaloid constituent was isolated from H. serrata (Thumb.) Trev..

CONCLUSIONThe compound was found to be a novel phlegmariurine type alkaloid, named 8 beta-hydroxy phlegmariurine B.

Heterocyclic Compounds, 3-Ring ; chemistry ; isolation & purification ; Huperzia ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry

A precise and selective reversed phase high-performance liquid chromatographic method (HPLC) was used to quantify the levels of huperzine A in samples of three Huperzia serrata populations with a total of 73 individuals located in Zhejiang, Guangxi, Chongqing, respectively, as well as in one-to-one samples of these 73 individuals introduced in same site after one year. Huperzine A content variation both among and within populations, and the dynamic change of this alkaloid occurring in same population after one year introduction, were analyzed using SPSS 13.0 software (Coefficient of variation, One-way ANOVA analysis, Paired-samples T tests). The results indicated that huperzine A content varied significantly by geographical locations, especially change with longitude, i. e., the order of the huperzine A content was CQ population > GX population > ZJ population. The coefficients of variation (CV) were as follows: 0.36 (CQ), 0.44 (GX) and 0.40 (ZJ). This indicated that there was plentiful diversity concerned with huperzine A content among individuals within population. Moreover, this high diversity was still maintained after one year introduction. ANOVA analysis showed that there was significant difference among populations in huperzine A content. Finally, the significant change of huperzine A content was not observed in all three populations after one year introduction. The results presented in this study could provide evidence that the huperzine A content variation of H. serrata is the results of an interaction between genes and the environment, by comparison, is mainly controlled by genetic factor.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; Huperzia ; chemistry

AIMTo study the alkaloid constituents of Huperzia serrata (Thunb.) Trev..

METHODSChromatographic methods were used for the isolation and purification. Structure was elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSAn alkaloid constituent was isolated from H. serrata (Thunb.) Trev..

CONCLUSIONThe compound was found to be a novel lycodine type alkaloid with tricyclic structure named huperzinine C.

Huperzia ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyridines ; chemistry ; isolation & purification

AIM: To investigate the chemical constituents from the whole plants of Phlegmariurus fargesii. METHOD: Compounds were isolated by repeated silica gel column chromatography. Their structures were elucidated by spectroscopic methods and chemical correlation. The acetylcholinesterase (AChE) inhibitory activity of the isolated compounds was evaluated. RESULTS: A new Lycopodium alkaloid, lycopodine N-oxide (1), along with lycopodine (2), 8,15-dehydrolycopodine (3), 6α-hydroxylycopodine (4), deacetyllycoclavine (5), N-methylhuperzine B (6), lycodine (7), and phlegmarine (8), was isolated. CONCLUSION Compound 1 is a new Lycopodium alkaloid, and compound 3 was obtained from nature for the first time. Other alkaloids are isolated from this plant for the first time.
Alkaloids ; chemistry ; isolation & purification ; Huperzia ; chemistry ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; Quinolizines ; chemistry ; isolation & purification

OBJECTIVETo investigate the constituents from Huperzia serrata.

METHODTwo diterpenoid compounds were isolated by column chromatography with silica gel and Sephadex LH-20. The structures were elucidated by spectral analysis and X-ray crystallographic analysis.

RESULTTheir structures were determined as (15R)-12, 16-epoxy-11, 14-dihydroxy-8, 11, 13-abietatrien-7-one (1) and 3beta-hydroxysandaracopimaric acid (2).

CONCLUSIONCompound 1 was a new natural abietane-type diterpenoid, compound 2 was isolated from this plant for the first time.

Crystallography, X-Ray ; Diterpenes ; analysis ; chemistry ; Huperzia ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Qiancengta, a traditional Chinese medicine produced from the whole plant of the club moss Huperzia serrata, has been used for a long history in China for treatment of a number of ailments, including contusions, strains, swellings, schizophrenia, myasthenia gravis and noworganophosphate poisoning. It has become known worldwide as a medicinal plant since Chinese scientists discovered huperzine A from it in the 1980s, which is reversible, potent and selective acetylcholine esterase (AChE) inhibitors by in vitro and in vivo pharmacological, and produce definite effects in the treatment of Alzheimer's disease. Now, Qiancengta is popular in all over the word for his famous pharmacological actions. For further exploitation this valuable resource under protection of nature environmental, its biological and ecological features, pharmaceutical active ingredients, artificial propagation and in vitro tissue culture, were reviewed, and the sustainable use of Qiancengta natural resource through plant biotechnology was put on the agenda.
Animals ; Biotechnology ; China ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Humans ; Huperzia ; chemistry ; physiology ; Plants, Medicinal ; chemistry ; physiology ; Research

A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.
Acetylcholinesterase ; Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Ascomycota ; chemistry ; classification ; isolation & purification ; Cholinesterase Inhibitors ; isolation & purification ; metabolism ; Endophytes ; classification ; isolation & purification ; Huperzia ; microbiology ; Mice ; RAW 264.7 Cells