Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid f...Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid fraction of the Chinese medicinal herb Huperzia serrata (Thunb.) Trev. Their structures and relative configurations were elucidated on the basis of spectroscopic analyses.展开更多
Huperzine V, a new Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and the absolute stereochemistry was determined by X-ray crystallographic analysis.
Huperzine W, a novel 14 carbons Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and its stucture was determined by spectroscopic analysis.
t Three new lycodine-type Lycopodium alkaloids,namely 1-methyllycodine(1),8a-hydroxy-15,16-dehydro-desN-methyl-a-obscurine(2),N-methyl-16-hydroxyhuperzine B(3),and one new natural lycodine-type Lycopodium alkaloid,N-...t Three new lycodine-type Lycopodium alkaloids,namely 1-methyllycodine(1),8a-hydroxy-15,16-dehydro-desN-methyl-a-obscurine(2),N-methyl-16-hydroxyhuperzine B(3),and one new natural lycodine-type Lycopodium alkaloid,N-methylhuperzine A(4),along with 11 known analogues(5–15),were isolated from the whole plants of club moss Huperzia serrata.The structures of 1–4 were elucidated on the basis of NMR spectroscopic and mass spectrometry data.Among them,compound 1 was the first lycodine-type alkaloid possessing a methyl group at C-1.In addition,the structure of 5 was confirmed by the single-crystal X-ray crystallography data and its^(13)C NMR was reported for the first time in current study.Compounds 1–5 were tested their BACE1 inhibitory activity.展开更多
To solve the problems of microbial contamination and no rooting during the tissue culture of Huperzia serrata, wild seedlings of Huperzia serrata were cultured by indoor hydroponics or soil cultivation methods, then t...To solve the problems of microbial contamination and no rooting during the tissue culture of Huperzia serrata, wild seedlings of Huperzia serrata were cultured by indoor hydroponics or soil cultivation methods, then the stem tips were used in tissue culture. The above operation significantly reduced microbial contamination during tissue culture process. Different types and concentrations of hormones were added into the basic medium MS to screen the optimal formula for induced plantlet growth and rooting. It was found that 1/2MSCO was the best medium on which stem, leaves and roots of tissue culture plantlets of Huperzia serrata grew quickly. At the same time, sand planting was a convenient and effective approach for gemmae propagation of Huperzia serrata, the survival rate of seedlings germinated from gemmae was nearly 100%.展开更多
[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate...[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate genes were analyzed using biogenetics technology.[Results]The molecular weight of the ARF7 proteins ranges from 11.28 to 608.54 kDa.HsARF7-4,HsARF7-5,HsARF7-8,HsARF7-9,HsARF7-10,HsARF7-13,HsARF7-16 and HsARF7-17 are basic proteins;all the proteins are unstable;except that HsARF7-6 and HsARF7-15 are hydrophilic,the rest are lipophilic proteins;there are 197 types of cis-acting elements,and 17 of the proteins contain as more as 20 acting elements such as GTGANTG10,CBFHV,GT1CONSENSUS,NODCON2GM,CAATBOX1,MYBCORE,GATABOX,WRKY71OS and RAV1AAT;HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-6,HsARF7-8,HsARF7-10,HsARF7-12,HsARF7-14 and HsARF7-16 all have the 6 types of motif predicted;the proteins have domains such as PABP-1234 super family,RRM_SF super famliy,PRK10263 super family,AUX_IAA super family and MFS super family;the proteins all can cross cell membrane and have no signal peptide,and they are all located in the nucleus.On the level of secondary structure,the proportion ofβ-sheet is the smallest.In HsARF7-7 and HsARF7-9,α-helix has the largest proportion,and in the remaining proteins,random coil has the largest proportion;and tertiary structure prediction found that HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-14 and HsARF7-17 have a tertiary structure of polyadenylate-binding protein,and HsARF7-7 has a tertiary structure of auxin-induced protein iaa4.[Conclusions]The above analysis results can provide a certain theoretical basis for further research on the biological function and regulatory mechanism of the ARF gene of H.serrata in the future,and provide a reference for the study of the growth and development of H.serrata.展开更多
Background:The male reproductive system consists of the testes,epididymis,accessory gonads,and genital ducts.The physiological integrity of the testicles is essential for successful sperm production,but the testes are...Background:The male reproductive system consists of the testes,epididymis,accessory gonads,and genital ducts.The physiological integrity of the testicles is essential for successful sperm production,but the testes are susceptible to a variety of exogenous and endogenous toxins,which can lead to structural damage to tissues and sperm abnormalities.Spermatogenesis is always in a homeostatic process and is sensitive to changes in the external environment.Huperzia serrata flavonoids(HSF)have a variety of pharmacological effects,including the treatment of bruises,strains,etc.HSF can improve the reproductive function of male mice,so the purpose of this experiment is to explore whether huperzine flavonoids are involved in and regulate the male reproductive process.Methods:Our experimental animals were ICR mice,and samples were collected after 28 days of HSF treatment,and verified by a series of experimental methods such as immunofluorescence,immunohistochemistry,HE staining,protein hybridization,and screening of related genes.Results:RNA sequencing results showed that HSF up-regulated genes related to sperm development and differentiation in testes and promoted spermatogenesis.HSF is effective in promoting steroid hormone synthesis by modulating STAR,CYP11A1,and CYP17A1.Conclusion:Thus,HSF can regulate testicular gene expression,stabilize the gut microbiome,alter serum metabolites,and regulate steroid homeostasis.展开更多
We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(HDR) gene family from Huperzia serrate.The two transcripts coding HDR,named Hs HDR1 and Hs HDR2,were discovered in t...We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(HDR) gene family from Huperzia serrate.The two transcripts coding HDR,named Hs HDR1 and Hs HDR2,were discovered in the transcriptome dataset of H.serrate and were cloned by reverse transcription-polymerase chain reaction(RT-PCR).The physicochemical properties,protein domains,protein secondary structure,and 3D structure of the putative Hs HDR1 and Hs HDR2 proteins were analyzed.The full-length c DNA of the Hs HDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids,whereas the Hs HDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids.These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(PF02401),but without the transmembrane region and signal peptide.The most abundant expression of Hs HDR1 and Hs HDR2 was detected in H.serrate roots,followed by the stems and leaves.Our results provide a foundation for exploring the function of Hs HDR1 and Hs HDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.展开更多
Thirteen triterpenoids, including three new ones, miyoshianols A (1), B (2) and C (3), were isolated from Huperzia miyoshiana . The structures of these new compounds were established as 3 O dihydroferuloyltoh...Thirteen triterpenoids, including three new ones, miyoshianols A (1), B (2) and C (3), were isolated from Huperzia miyoshiana . The structures of these new compounds were established as 3 O dihydroferuloyltohogenol (1) , 16 oxo 3 β ,21 β dihydroxy serrat 14 en 24 ferulate (2) and 16 oxo 3 α ,21 β dihydroxy serrat 14 en 24 ferulate (3), respectively, on the basis of their spectroscopic analysis.展开更多
Qian ceng Ta,the whole plant of Huperzia serrata,is an important landscape and medicinal herbs and contains abundant bioactive lycopodium alkaloids.Although the structures of more than 100 lycopodium alkaloids in Hupe...Qian ceng Ta,the whole plant of Huperzia serrata,is an important landscape and medicinal herbs and contains abundant bioactive lycopodium alkaloids.Although the structures of more than 100 lycopodium alkaloids in Huperzia serrata have been isolated and identified,the content and distribution of these alkaloids in different tissues are still unclear.In current study,an ultra-performance liquid chromatography-mass spectrometry based comprehensive metabolomics strategy was developed,including the extraction,separation,identification,and statistical analysis.The results showed that different types lycopodium alkaloids could be separated at different time-windows,which was helpful for further metabolite identification.Peak4388 and peak3954 were metabolite biomarkers for the different tissues according to the principle component analysis and partial least squares-discriminant analysis model.A computational tool based in-house database was also built up and used for putative identification.Of the 2354 true peaks after four-step filtration,118 peaks were putatively identified as lycopodium alkaloids by using in-house database,and four of which was identified by authentic standards.Alternatively,another computational software was used to predict the fragmentation pattern,to dereplicate the structure of identified peaks,and identified the peak3585 to N-methylhuperzine A.The integration of both computational tools could be used for more metabolites identification.展开更多
A new Lycopodium alkaloid together with a phenolic glycoside—arbutin, were isolated from the whole plant of Huperzia serrata. The new compound was assigned the trivial name of huperzine I. The structures of these two...A new Lycopodium alkaloid together with a phenolic glycoside—arbutin, were isolated from the whole plant of Huperzia serrata. The new compound was assigned the trivial name of huperzine I. The structures of these two compounds were determined on the basis of spectral evidence.展开更多
文摘Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid fraction of the Chinese medicinal herb Huperzia serrata (Thunb.) Trev. Their structures and relative configurations were elucidated on the basis of spectroscopic analyses.
文摘Huperzine V, a new Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and the absolute stereochemistry was determined by X-ray crystallographic analysis.
文摘Huperzine W, a novel 14 carbons Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and its stucture was determined by spectroscopic analysis.
基金the NSFC-Joint Foundation of Yunnan Province(No.U1502223)the National Natural Science Foundation of China(No.21402212)+1 种基金the Science and Technology Program of Yunnan province(No 2015FB173)the CAS"Light of West China"Program and Youth Innovation Promotion Association CAS(X.D.Wu).
文摘t Three new lycodine-type Lycopodium alkaloids,namely 1-methyllycodine(1),8a-hydroxy-15,16-dehydro-desN-methyl-a-obscurine(2),N-methyl-16-hydroxyhuperzine B(3),and one new natural lycodine-type Lycopodium alkaloid,N-methylhuperzine A(4),along with 11 known analogues(5–15),were isolated from the whole plants of club moss Huperzia serrata.The structures of 1–4 were elucidated on the basis of NMR spectroscopic and mass spectrometry data.Among them,compound 1 was the first lycodine-type alkaloid possessing a methyl group at C-1.In addition,the structure of 5 was confirmed by the single-crystal X-ray crystallography data and its^(13)C NMR was reported for the first time in current study.Compounds 1–5 were tested their BACE1 inhibitory activity.
文摘To solve the problems of microbial contamination and no rooting during the tissue culture of Huperzia serrata, wild seedlings of Huperzia serrata were cultured by indoor hydroponics or soil cultivation methods, then the stem tips were used in tissue culture. The above operation significantly reduced microbial contamination during tissue culture process. Different types and concentrations of hormones were added into the basic medium MS to screen the optimal formula for induced plantlet growth and rooting. It was found that 1/2MSCO was the best medium on which stem, leaves and roots of tissue culture plantlets of Huperzia serrata grew quickly. At the same time, sand planting was a convenient and effective approach for gemmae propagation of Huperzia serrata, the survival rate of seedlings germinated from gemmae was nearly 100%.
基金High-level Innovation Team Project of Colleges and Universities in Guangxi(Gui Jiao 2019-52)Natural Science Foundation of Guangxi(2017GXNSFBA198094)+5 种基金Basic Ability Enhancement Project of Education Department of Guangxi for Young and Middle-aged Teachers(2017KY495)Scientific Research Project of Guangxi University of Chinese Medicine(2016QN013)Talent Training and Construction Project of Professor Yang Shilin's Team(YSL17026)Special Project of Guangxi University of Chinese Medicine for Construction of Superior Subject of Traditional Chinese Medicine(CICAR 2016-P4)Science and Technology Development Fund of Guangxi Academy of Agricultural Sciences(Gui Nong Ke 2018JZ36)High-level Talent Team under Qihuang Project of Guangxi University of Chinese Medicine(2018002).
文摘[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate genes were analyzed using biogenetics technology.[Results]The molecular weight of the ARF7 proteins ranges from 11.28 to 608.54 kDa.HsARF7-4,HsARF7-5,HsARF7-8,HsARF7-9,HsARF7-10,HsARF7-13,HsARF7-16 and HsARF7-17 are basic proteins;all the proteins are unstable;except that HsARF7-6 and HsARF7-15 are hydrophilic,the rest are lipophilic proteins;there are 197 types of cis-acting elements,and 17 of the proteins contain as more as 20 acting elements such as GTGANTG10,CBFHV,GT1CONSENSUS,NODCON2GM,CAATBOX1,MYBCORE,GATABOX,WRKY71OS and RAV1AAT;HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-6,HsARF7-8,HsARF7-10,HsARF7-12,HsARF7-14 and HsARF7-16 all have the 6 types of motif predicted;the proteins have domains such as PABP-1234 super family,RRM_SF super famliy,PRK10263 super family,AUX_IAA super family and MFS super family;the proteins all can cross cell membrane and have no signal peptide,and they are all located in the nucleus.On the level of secondary structure,the proportion ofβ-sheet is the smallest.In HsARF7-7 and HsARF7-9,α-helix has the largest proportion,and in the remaining proteins,random coil has the largest proportion;and tertiary structure prediction found that HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-14 and HsARF7-17 have a tertiary structure of polyadenylate-binding protein,and HsARF7-7 has a tertiary structure of auxin-induced protein iaa4.[Conclusions]The above analysis results can provide a certain theoretical basis for further research on the biological function and regulatory mechanism of the ARF gene of H.serrata in the future,and provide a reference for the study of the growth and development of H.serrata.
基金supported by a high-level talent research fund project of Qingdao Agricultural University in China(1120043).
文摘Background:The male reproductive system consists of the testes,epididymis,accessory gonads,and genital ducts.The physiological integrity of the testicles is essential for successful sperm production,but the testes are susceptible to a variety of exogenous and endogenous toxins,which can lead to structural damage to tissues and sperm abnormalities.Spermatogenesis is always in a homeostatic process and is sensitive to changes in the external environment.Huperzia serrata flavonoids(HSF)have a variety of pharmacological effects,including the treatment of bruises,strains,etc.HSF can improve the reproductive function of male mice,so the purpose of this experiment is to explore whether huperzine flavonoids are involved in and regulate the male reproductive process.Methods:Our experimental animals were ICR mice,and samples were collected after 28 days of HSF treatment,and verified by a series of experimental methods such as immunofluorescence,immunohistochemistry,HE staining,protein hybridization,and screening of related genes.Results:RNA sequencing results showed that HSF up-regulated genes related to sperm development and differentiation in testes and promoted spermatogenesis.HSF is effective in promoting steroid hormone synthesis by modulating STAR,CYP11A1,and CYP17A1.Conclusion:Thus,HSF can regulate testicular gene expression,stabilize the gut microbiome,alter serum metabolites,and regulate steroid homeostasis.
基金supported by the National Nature Science Foundation of China(No.30900113)the Program for Innovative Research Team in the Institute of Medicinal Plant Development(No.IT1304)
文摘We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(HDR) gene family from Huperzia serrate.The two transcripts coding HDR,named Hs HDR1 and Hs HDR2,were discovered in the transcriptome dataset of H.serrate and were cloned by reverse transcription-polymerase chain reaction(RT-PCR).The physicochemical properties,protein domains,protein secondary structure,and 3D structure of the putative Hs HDR1 and Hs HDR2 proteins were analyzed.The full-length c DNA of the Hs HDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids,whereas the Hs HDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids.These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(PF02401),but without the transmembrane region and signal peptide.The most abundant expression of Hs HDR1 and Hs HDR2 was detected in H.serrate roots,followed by the stems and leaves.Our results provide a foundation for exploring the function of Hs HDR1 and Hs HDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.
基金ProjectsupportedbytheNationalNaturalScienceFoundationofChina (No .3 990 0 0 13 )
文摘Thirteen triterpenoids, including three new ones, miyoshianols A (1), B (2) and C (3), were isolated from Huperzia miyoshiana . The structures of these new compounds were established as 3 O dihydroferuloyltohogenol (1) , 16 oxo 3 β ,21 β dihydroxy serrat 14 en 24 ferulate (2) and 16 oxo 3 α ,21 β dihydroxy serrat 14 en 24 ferulate (3), respectively, on the basis of their spectroscopic analysis.
基金This work was financially supported by the Science and Technology Commission of Shanghai Municipality(Grant 15JC1400402)CAS-JIC Centre of Excellence in Plant and Microbial Sciences(CEPAMS)funding,and the Strategic Priority Research Program“Molecular mechanism of Plant Growth and Development”of CAS(Grant XDPB0402).
文摘Qian ceng Ta,the whole plant of Huperzia serrata,is an important landscape and medicinal herbs and contains abundant bioactive lycopodium alkaloids.Although the structures of more than 100 lycopodium alkaloids in Huperzia serrata have been isolated and identified,the content and distribution of these alkaloids in different tissues are still unclear.In current study,an ultra-performance liquid chromatography-mass spectrometry based comprehensive metabolomics strategy was developed,including the extraction,separation,identification,and statistical analysis.The results showed that different types lycopodium alkaloids could be separated at different time-windows,which was helpful for further metabolite identification.Peak4388 and peak3954 were metabolite biomarkers for the different tissues according to the principle component analysis and partial least squares-discriminant analysis model.A computational tool based in-house database was also built up and used for putative identification.Of the 2354 true peaks after four-step filtration,118 peaks were putatively identified as lycopodium alkaloids by using in-house database,and four of which was identified by authentic standards.Alternatively,another computational software was used to predict the fragmentation pattern,to dereplicate the structure of identified peaks,and identified the peak3585 to N-methylhuperzine A.The integration of both computational tools could be used for more metabolites identification.
文摘A new Lycopodium alkaloid together with a phenolic glycoside—arbutin, were isolated from the whole plant of Huperzia serrata. The new compound was assigned the trivial name of huperzine I. The structures of these two compounds were determined on the basis of spectral evidence.
