The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Dayuan Zhu - One of the best experts on this subject based on the ideXlab platform.
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lycopodium alkaloids from Huperzia serrata
Journal of Asian Natural Products Research, 2010Co-Authors: Yanfang Yang, Changheng Tan, Junjie Tan, Shanhao Jiang, Kai Xiao, Dayuan ZhuAbstract:A new lycopodane-type Lycopodium alkaloid, 6α-hydroxy-5,15-oxide-lycopodane (1), and seven known alkaloids were isolated from the whole plants of Huperzia serrata. Their structures were elucidated by means of spectroscopic methods. 12-Deoxyhuperzine O (2) was reported as a naturally occurring alkaloid for the first time, and showed an antagonist effect on the N-methyl-d-aspartate receptor with an IC50 value of 0.92 μM.
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two new n oxide lycopodium alkaloids from Huperzia serrata
Natural Product Research, 2009Co-Authors: Hengbin Wang, Changheng Tan, Junjie Tan, Shanhao Jiang, Yilei Chen, Dayuan ZhuAbstract:Two new Lycopodium alkaloids, N-oxidehuperzine E (1) and N-oxidehuperzine F (2), along with two known alkaloids, huperzines E (3) and F (4), were isolated from Huperzia serrata (Thunb.) Trev. Their structures were elucidated by spectroscopic and chemical transformations.
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two new nitrone alkaloids from Huperzia serrata
Helvetica Chimica Acta, 2008Co-Authors: Wenyun Gao, Shanhao Jiang, Dayuan ZhuAbstract:Two new nitrone alkaloids were isolated from the whole plant of Huperzia serrata (Thunb.) Trev. They are both phlegmarine-type lycopodium alkaloids with a nitrone moiety. Their structures were elucidated on the basis of spectral evidences, and their configurations were established on the basis of optical rotation, CD, and NOESY-NMR data.
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lycopodium alkaloids from Huperzia serrata
ChemInform, 2007Co-Authors: Hengbin Wang, Changheng Tan, Junjie Tan, Mingyao Gao, Shanhao Jiang, Dayuan ZhuAbstract:Three new Lycopodium alkaloids, 2-chlorohuperzine E (1), huperzines E′ (2), and F′ (3), along with two known compounds, huperzines E and F, were isolated from Huperzia serrata (Thunb.) Trev. Their structures were elucidated by spectroscopic methods.
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is there a better source of huperzine a than Huperzia serrata huperzine a content of Huperziaceae species in china
Journal of Agricultural and Food Chemistry, 2005Co-Authors: Changheng Tan, Dayuan Zhu, David R GangAbstract:A precise and selective reversed phase high-performance liquid chromatographic method was developed for quantifying huperzine A (HupA) in samples of the Huperziaceae in China. This method was used to quantify the levels of HupA in samples of Huperzia serrata collected from a single population at different times of the year, in different organs of the same H. serrata plant, and from different geographical locations of H. serrata plants in China. For different species of Huperziaceae, the highest content of HupA was found in Phlegmariurus carinatus. Members of the genus Phlegmariurus possessed higher levels of HupA than Huperzia species. H. serrata plants growing in humid forests contained significantly more HupA than plants growing in less humid environments. Finally, HupA content varied significantly by season, with the highest levels being found in mid fall and the lowest levels in early spring, suggesting that HupA is turned over in the plant.
Zhajun Zhan - One of the best experts on this subject based on the ideXlab platform.
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biotransformation of huperzine a by irpex lacteus a fungal endophyte of Huperzia serrata
Fitoterapia, 2019Co-Authors: Youmin Ying, Weiguang Shan, Cuiping Tong, Caixue Zhang, Zhidong Zhang, Qiyong Tang, Wei Mao, Yun Zhang, Zhajun ZhanAbstract:Abstract The biotransformation of huperzine A (hupA), one of the characteristic bioactive constituents of the medicinal plant Huperzia serrata, by a fungal endophyte of the host plant was studied. Two previously undescribed compounds 1–2, along with a known analog 8α,15α-epoxyhuperzine A (3), were isolated and identified. The structures of all the isolates were established by spectroscopic methods including NMR, MS, IR, and UV spectra. In particular, the absolute configurations of 1 and 2 were elucidated by CD spectra comparison and theoretic NOE strength calculation. In the LPS-induced neuro-inflammation injury assay, 1–3 exhibited moderate neuroprotective activity by increasing the viability of U251 cell lines with EC50 values of 35.3 ± 0.9, 32.1 ± 0.9, and 50.3 ± 0.8 nM, respectively.
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biotransformation of huperzine b by a fungal endophyte of Huperzia serrata
Chemistry & Biodiversity, 2019Co-Authors: Zhajun Zhan, Weiguang Shan, Ting Tian, Caixue Zhang, Zhidong Zhang, Qiyong Tang, Youmin YingAbstract:The biotransformation of huperzine B (hupB), one of the characteristic bioactive constituents of the medicinal plant Huperzia serrata, by a fungal endophyte of the host plant was studied. One new compound, 8α,15α-epoxyhuperzine B (1), along with two known oxygenated hupB analogs, 16-hydroxyhuperzine B (2) and carinatumin B (3), was isolated and identified. The structures of all the isolates were deduced by spectroscopic methods including NMR, MS, IR, and UV spectra. The known compounds 2 and 3 were obtained from a microbial source for the first time. To the best of our knowledge, it is the first report on the microbial transformation of hupB and would facilitate further structural modification of hupB by chemo-enzymatic method. In the LPS-induced neuro-inflammation injury assay, 8α,15α-epoxyhuperzine B (1) exhibited moderate neuroprotective activity by increasing the viability of U251 cell lines with an EC50 of 40.1 nm.
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lycopodium alkaloids from Huperzia serrata
ChemInform, 2015Co-Authors: Youmin Ying, Cuiping Tong, Zhajun Zhan, Xiaosa Liu, Jianwei Wang, Weiguang ShanAbstract:isolation, structure elucidation, AChE- and α-glucosidase inhibitory activities of three new lycopodium alkaloids, huperserramines A—C (I)—(III) along with 15 known related compounds
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xanthones with α glucosidase inhibitory activities from aspergillus versicolor a fungal endophyte of Huperzia serrata
Helvetica Chimica Acta, 2015Co-Authors: Weiguang Shan, Youmin Ying, Wenhong Liu, Zhajun ZhanAbstract:Three new xanthones, namely huperxanthones A–C (1–3, resp.), were obtained from the cultures of Aspergillus versicolor, a fungal endophyte of Huperzia serrata, together with 1,7-dihydroxy-8-(methoxycarbonyl)xanthone-3-carboxylic acid (4), β-diversonolic acid methyl ester (5), 4-hydroxyvertixanthone (6), and sydowinin B (7). The structures of the new compounds were established by detailed NMR and MS analysis, especially by 2D-NMR experiments. All xanthones were evaluated for their effects on α-glucosidase. Compound 4 exhibited a potent inhibitory activity against α-glucosidase with an IC50 value of 0.24 mM (vs. 0.38 mM for acarbose). The rest of the compounds showed weak or no activity against α-glucosidase.
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lycopodium alkaloids from Huperzia serrata
Helvetica Chimica Acta, 2014Co-Authors: Youmin Ying, Cuiping Tong, Zhajun Zhan, Xiaosa Liu, Jianwei Wang, Weiguang ShanAbstract:Three new lycopodium alkaloids, huperserramines A–C (1–3, resp.), along with 15 known ones, lycopodine-6α,11α-diol (4), lycoposerramine H (5), lycoposerramine I (6), lycopodine-6α-ol (7), lycoposerramine M (8), diphaladine A (9), lycoposerramine K (10), lycoposerramine W (11), huperzine M (12), luciduline (13), phlegmariuine N (14), huperzine A (15), huperzine B (16), lycodine (17), and lycoposerramine R (18), were isolated from the whole plant of Huperzia serrata. Their structures were established by spectroscopic methods, including 2D-NMR and MS analyses. All the isolates were evaluated for their inhibitory effects on acetylcholinesterase (AChE) and α-glucosidase. As a result, lycopodine-6α,11α-diol (4) exhibited more potent α-glucosidase inhibitory activity (IC50 148±5.5 μM) than the positive control acarbose (IC50 376.3±2.7 μM).
Shepo Shi - One of the best experts on this subject based on the ideXlab platform.
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enzymatic synthesis of 2 hydroxy 4h quinolizin 4 one scaffolds by integrating coenzyme a ligases and a type iii pks from Huperzia serrata
RSC Advances, 2020Co-Authors: Juan Wang, Ning Ding, Xiao Liu, Xiaohui Wang, Xiaoping Shi, Shepo ShiAbstract:2-Hydroxy-4H-quinolizin-4-one scaffolds were enzymatically synthesized by integrating three enzymes including phenylacetate-CoA ligase (PcPCL) from an endophytic fungus Penicillium chrysogenum MT-12, malonyl-CoA synthase (AtMatB) from Arabidopsis thaliana, and a type III polyketide synthase (HsPKS3) from Chinese club moss Huperzia serrata. The findings paved the way to produce these kinds of structurally interesting alkaloids by engineered microorganisms.
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lycopodium alkaloids from Huperzia serrata
Fitoterapia, 2019Co-Authors: Fangfang Jiang, Ning Ding, Hongyun Yang, Fangfang Jia, Yuan Luo, Juan Wang, Xiao Liu, Xiaohui Wang, Shepo ShiAbstract:Abstract Five new Lycopodium alkaloids, huperzine Y1 (1), huperzine Y2 (2), huperzine Y3 (3), (+)-huperzine Z (4a) and (−)-huperzine Z (4b) as well as ten known alkaloids (5–14) were isolated from Huperzia serrata. The structures of the new compounds were established using extensive spectroscopic (1D and 2D NMR, IR, and HRESIMS) and calculated electronic circular dichroism (ECD) methods. Compounds 4a and 4b were a pair of enantiomers successfully separated by chiral HPLC resolution. Compounds 2 and 3 indicated inhibitory activities against acetylcholinesterase with IC50 value of 57.1 ± 1.6 and 32.7 ± 1.0 μΜ, respectively. However, no compound showed inhibitory effect on butyrocholinesterase at the concentration of 100 μΜ.
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polyketides from alternaria alternata mt 47 an endophytic fungus isolated from Huperzia serrata
Fitoterapia, 2019Co-Authors: Hongyun Yang, Fangfang Jiang, Ning Ding, Fangfang Jia, Yuan Luo, Xiao Liu, Lili Wang, Zhixiang Zhu, Shepo ShiAbstract:Abstract Four new polyketides, alternatains A-D (1–4), along with 17 known compounds (5–21) were obtained from the solid substrate fermentation cultures of Alternaria alternata MT-47, an endophytic fungus isolated from the medicinal plant of Huperzia serrata. Their structures were elucidated by extensive spectroscopic and spectrometric techniques (1D and 2D NMR, IR, and HRESIMS) and calculated electronic circular dichroism (ECD) method. Compounds 4, 6, 15, and 21 exhibited inhibitory activities on ATP release of thrombin-activated platelets with IC50 values in the range of 18.2–68.8 μM.
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Isolation and identification of endophytic fungi from Huperzia serrata and their metabolites' inhibitory activities against acetylcholinesterase and anti-inflammatory activities
Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica, 2019Co-Authors: Xin Zhang, Hongyun Yang, Xiaohui Wang, Shepo Shi, Yaru Yan, Xiao LiuAbstract: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.
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nitric oxide inhibitory polyketides from penicillium chrysogenum mt 12 an endophytic fungus isolated from Huperzia serrata
Fitoterapia, 2017Co-Authors: Xiao Liu, Juan Wang, Xiaohui Wang, Zhixiang Zhu, Xiaoping Shi, Yunfang Zhao, Hongwei Jin, Shepo ShiAbstract:Twelve new polyketides, penicichrysogenins A-L (1-10, 11a, and 11b) along with five known compounds (12a, 12b, and 13-15) were isolated from the solid substrate fermentation cultures of a Huperzia serrata endophytic fungus Penicillium chrysogenum MT-12. The structures of the new compounds were established using extensive spectroscopic (1D and 2D NMR, IR, and HRESIMS) and calculated electronic circular dichroism (ECD) methods. Compounds 11a/11b and 12a/12b were two pairs of enantiomers successfully separated by chiral HPLC resolution. Compounds 4, 5, 8, 9, 11a/11b, and 12a/12b exhibited inhibition of nitric oxide production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells with IC50 values in the range of 17.5-98.4μM.
Youli Xiao - One of the best experts on this subject based on the ideXlab platform.
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Selection of Reference Genes for Expression Analysis in Chinese Medicinal Herb Huperzia serrata.
Frontiers in Pharmacology, 2019Co-Authors: Mengquan Yang, Wenjing You, Amit Jaisi, Youli XiaoAbstract:Huperzine A (HupA) is a powerful and selective inhibitor of acetylcholinesterase. It has attracted widespread attention endangering the ultimate plant sources of Lycopodiaceae family. In this study, we used Huperzia serrata, extensively used in Traditional Chinese medicine (TCM), a slow growing vascular plant as the model plant of the Lycopodiaceae family to develop and validate the reference genes. We aim to use gene expression platform to understand the gene expression of different tissues and developmental stages of this medicinal herb. Eight candidate reference genes were selected based on RNA-seq data and evaluated with qRT-PCR. The expression of L/ODC and cytochrome P450s genes known for their involvement in lycopodium alkaloid biosynthesis, were also studied to validate the selected reference genes. The most stable genes were TBP, GAPDH, and their combination (TBP + GAPDH). We report for the first time the reference gene of H. serrata’s different tissues which would provide important insights into understanding their biological functions comparing other Lycopodiaceae plants and facilitate a good biopharming approach.
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Table_1_Selection of Reference Genes for Expression Analysis in Chinese Medicinal Herb Huperzia serrata.XLSX
2019Co-Authors: Mengquan Yang, Wenjing You, Amit Jaisi, Youli XiaoAbstract:Huperzine A (HupA) is a powerful and selective inhibitor of acetylcholinesterase. It has attracted widespread attention endangering the ultimate plant sources of Lycopodiaceae family. In this study, we used Huperzia serrata, extensively used in Traditional Chinese medicine (TCM), a slow growing vascular plant as the model plant of the Lycopodiaceae family to develop and validate the reference genes. We aim to use gene expression platform to understand the gene expression of different tissues and developmental stages of this medicinal herb. Eight candidate reference genes were selected based on RNA-seq data and evaluated with qRT-PCR. The expression of L/ODC and cytochrome P450s genes known for their involvement in lycopodium alkaloid biosynthesis, were also studied to validate the selected reference genes. The most stable genes were TBP, GAPDH, and their combination (TBP + GAPDH). We report for the first time the reference gene of H. serrata’s different tissues which would provide important insights into understanding their biological functions comparing other Lycopodiaceae plants and facilitate a good biopharming approach.
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insights into pipecolic acid biosynthesis in Huperzia serrata
Organic Letters, 2018Co-Authors: Zhen Fan, Amit Jaisi, Yongxing Lei, Yu Ping, Youli XiaoAbstract:For the biosynthesis of Pip in Huperzia serrata, the mechanistic studies were evaluated. Through a series of biochemical analyses, Pip is biosynthesized through a two-step cascade reaction. Three intermediates possibly exist simultaneously as an equilibrium matter in the first-step reaction catalyzed by HsAld1, while HsSard4 performs as a ketimine reductase and chemoselectively and stereoselectively takes 1,2-dehydropipecolic acid as the preferred substrate in vitro.
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comprehensive relative quantitative metabolomics analysis of lycopodium alkaloids in different tissues of Huperzia serrata
Synthetic and Systems Biotechnology, 2017Co-Authors: Zhen Fan, Youli XiaoAbstract: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.
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global transcriptome analysis of Huperzia serrata and identification of critical genes involved in the biosynthesis of huperzine a
BMC Genomics, 2017Co-Authors: Mengquan Yang, Wenjing You, Zhen Fan, Mulan Zhu, Youli XiaoAbstract:Huperzia serrata (H. serrata) is an economically important traditional Chinese herb with the notably medicinal value. As a representative member of the Lycopodiaceae family, the H. serrata produces various types of effectively bioactive lycopodium alkaloids, especially the huperzine A (HupA) which is a promising drug for Alzheimer’s disease. Despite their medicinal importance, the public genomic and transcriptomic resources are very limited and the biosynthesis of HupA is largely unknown. Previous studies on comparison of 454-ESTs from H. serrata and Phlegmariurus carinatus predicted putative genes involved in lycopodium alkaloid biosynthesis, such as lysine decarboxylase like (LDC-like) protein and some CYP450s. However, these gene annotations were not carried out with further biochemical characterizations. To understand the biosynthesis of HupA and its regulation in H. serrata, a global transcriptome analysis on H. serrata tissues was performed. In this study, we used the Illumina Highseq4000 platform to generate a substantial RNA sequencing dataset of H. serrata. A total of 40.1 Gb clean data was generated from four different tissues: root, stem, leaf, and sporangia and assembled into 181,141 unigenes. The total length, average length, N50 and GC content of unigenes were 219,520,611 bp, 1,211 bp, 2,488 bp and 42.51%, respectively. Among them, 105,516 unigenes (58.25%) were annotated by seven public databases (NR, NT, Swiss-Prot, KEGG, COG, Interpro, GO), and 54 GO terms and 3,391 transcription factors (TFs) were functionally classified, respectively. KEGG pathway analysis revealed that 72,230 unigenes were classified into 21 functional pathways. Three types of candidate enzymes, LDC, CAO and PKS, responsible for the biosynthesis of precursors of HupA were all identified in the transcripts. Four hundred and fifty-seven CYP450 genes in H. serrata were also analyzed and compared with tissue-specific gene expression. Moreover, two key classes of CYP450 genes BBE and SLS, with 23 members in total, for modification of the lycopodium alkaloid scaffold in the late two stages of biosynthesis of HupA were further evaluated. This study is the first report of global transcriptome analysis on all tissues of H. serrata, and critical genes involved in the biosynthesis of precursors and scaffold modifications of HupA were discovered and predicted. The transcriptome data from this work not only could provide an important resource for further investigating on metabolic pathways in H. serrata, but also shed light on synthetic biology study of HupA.
Shilin Chen - One of the best experts on this subject based on the ideXlab platform.
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Cloning and analysis of squalene synthase (HsSQS1) gene in Huperzia serrata
Yao xue xue bao = Acta pharmaceutica Sinica, 2012Co-Authors: Xiu-mei Yin, Zhi-chuan Bai, Yunyun Niu, Hongmei Luo, Shilin Chen, Shilin ChenAbstract:Squalene synthase (SQS) is a key enzyme in plant terpenoid biosynthetic pathway. This study focused on cloning and analysis of Huperzia serrata SQS (HsSQS1) gene. After searching the transcriptome dataset of H serrata, one unique sequence encoding SQS was discovered. The primers were designed according to the transcript sequence of HsSQS1 from the H. serrata transcriptome dataset. The open reading frame of HsSQS1 was cloned using RT-PCR strategy. The bioinformatic analysis of this gene and its corresponding protein were performed. The cDNA (named as HsSQS1) contains a 1263 bp open reading frame and encodes a predicted protein of 420 amino acids. The GenBank accession number for this gene is JQ004938. HsSQS1 contains two transmembrane regions, without signal peptide. The conserved domain of squalene synthase was presented in HsSQS1. HsSQS1 was more abundant in H. serrata root than in leaf and stem. This study cloned and analyzed squalene synthase gene from H. serrata for the first time. The result will provide a foundation for exploring the mechanism ofterpenoid biosynthesis in H. serrata plants.
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comparison of 454 ests from Huperzia serrata and phlegmariurus carinatus reveals putative genes involved in lycopodium alkaloid biosynthesis and developmental regulation
BMC Plant Biology, 2010Co-Authors: Ying Li, Jingyuan Song, Qiong Wu, Andre Steinmetz, Shilin ChenAbstract:Background: Plants of the Huperziaceae family, which comprise the two genera Huperzia and Phlegmariurus, produce various types of lycopodium alkaloids that are used to treat a number of human ailments, such as contusions, swellings and strains. Huperzine A, which belongs to the lycodine type of lycopodium alkaloids, has been used as an anti-Alzheimer’s disease drug candidate. Despite their medical importance, little genomic or transcriptomic data are available for the members of this family. We used massive parallel pyrosequencing on the Roche 454-GS FLX Titanium platform to generate a substantial EST dataset for Huperzia serrata (H. serrata) and Phlegmariurus carinatus (P. carinatus) as representative members of the Huperzia and Phlegmariurus genera, respectively. H. serrata and P. carinatus are important plants for research on the biosynthesis of lycopodium alkaloids. We focused on gene discovery in the areas of bioactive compound biosynthesis and transcriptional regulation as well as genetic marker detection in these species. Results: For H. serrata, 36,763 unique putative transcripts were generated from 140,930 reads totaling over 57,028,559 base pairs; for P. carinatus, 31,812 unique putative transcripts were generated from 79,920 reads totaling over 30,498,684 base pairs. Using BLASTX searches of public databases, 16,274 (44.3%) unique putative transcripts from H. serrata and 14,070 (44.2%) from P. carinatus were assigned to at least one protein. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology annotations revealed that the functions of the unique putative transcripts from these two species cover a similarly broad set of molecular functions, biological processes and biochemical pathways. In particular, a total of 20 H. serrata candidate cytochrome P450 genes, which are more abundant in leaves than in roots and might be involved in lycopodium alkaloid biosynthesis, were found based on the comparison of H. serrata and P. carinatus 454-ESTs and real-time PCR analysis. Four unique putative CYP450 transcripts (Hs01891, Hs04010, Hs13557 and Hs00093) which are the most likely to be involved in the biosynthesis of lycopodium alkaloids were selected based on a phylogenetic analysis. Approximately 115 H. serrata and 98 P. carinatus unique putative transcripts associated with the biosynthesis of triterpenoids, alkaloids and flavones/flavonoids were located in the 454-EST datasets. Transcripts related to phytohormone biosynthesis and signal transduction as well as transcription factors were also obtained. In addition, we discovered 2,729 and 1,573 potential SSR-motif microsatellite loci in the H. serrata and P. carinatus 454-ESTs, respectively.
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comparison of 454 ests from Huperzia serrata and phlegmariurus carinatus reveals putative genes involved in lycopodium alkaloid biosynthesis and developmental regulation
BMC Plant Biology, 2010Co-Authors: Hongmei Luo, Chao Sun, Jingyuan Song, Andre Steinmetz, Yongzhen Sun, Shilin ChenAbstract:Plants of the Huperziaceae family, which comprise the two genera Huperzia and Phlegmariurus, produce various types of lycopodium alkaloids that are used to treat a number of human ailments, such as contusions, swellings and strains. Huperzine A, which belongs to the lycodine type of lycopodium alkaloids, has been used as an anti-Alzheimer's disease drug candidate. Despite their medical importance, little genomic or transcriptomic data are available for the members of this family. We used massive parallel pyrosequencing on the Roche 454-GS FLX Titanium platform to generate a substantial EST dataset for Huperzia serrata (H. serrata) and Phlegmariurus carinatus (P. carinatus) as representative members of the Huperzia and Phlegmariurus genera, respectively. H. serrata and P. carinatus are important plants for research on the biosynthesis of lycopodium alkaloids. We focused on gene discovery in the areas of bioactive compound biosynthesis and transcriptional regulation as well as genetic marker detection in these species. For H. serrata, 36,763 unique putative transcripts were generated from 140,930 reads totaling over 57,028,559 base pairs; for P. carinatus, 31,812 unique putative transcripts were generated from 79,920 reads totaling over 30,498,684 base pairs. Using BLASTX searches of public databases, 16,274 (44.3%) unique putative transcripts from H. serrata and 14,070 (44.2%) from P. carinatus were assigned to at least one protein. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology annotations revealed that the functions of the unique putative transcripts from these two species cover a similarly broad set of molecular functions, biological processes and biochemical pathways. In particular, a total of 20 H. serrata candidate cytochrome P450 genes, which are more abundant in leaves than in roots and might be involved in lycopodium alkaloid biosynthesis, were found based on the comparison of H. serrata and P. carinatus 454-ESTs and real-time PCR analysis. Four unique putative CYP450 transcripts (Hs01891, Hs04010, Hs13557 and Hs00093) which are the most likely to be involved in the biosynthesis of lycopodium alkaloids were selected based on a phylogenetic analysis. Approximately 115 H. serrata and 98 P. carinatus unique putative transcripts associated with the biosynthesis of triterpenoids, alkaloids and flavones/flavonoids were located in the 454-EST datasets. Transcripts related to phytohormone biosynthesis and signal transduction as well as transcription factors were also obtained. In addition, we discovered 2,729 and 1,573 potential SSR-motif microsatellite loci in the H. serrata and P. carinatus 454-ESTs, respectively. The 454-EST resource allowed for the first large-scale acquisition of ESTs from H. serrata and P. carinatus, which are representative members of the Huperziaceae family. We discovered many genes likely to be involved in the biosynthesis of bioactive compounds and transcriptional regulation as well as a large number of potential microsatellite markers. These results constitute an essential resource for understanding the molecular basis of developmental regulation and secondary metabolite biosynthesis (especially that of lycopodium alkaloids) in the Huperziaceae, and they provide an overview of the genetic diversity of this family.
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analysis of expressed sequence tags from the Huperzia serrata leaf for gene discovery in the areas of secondary metabolite biosynthesis and development regulation
Physiologia Plantarum, 2010Co-Authors: Hongmei Luo, Deli Wang, Chao Sun, Jingyuan Song, Xiaocheng Jia, Shilin ChenAbstract:Huperzia serrata produces various types of lycopodium alkaloids, especially the huperzine A (HupA) that is a promising drug candidate for Alzheimer's disease. Despite the medicinal importance of H. serrata, little genomic or transcriptomic data are available from the public databases. A cDNA library was thus generated from RNA isolated from the leaves of H. serrata. A total of 4012 clones were randomly selected from the library, and 3451 high-quality expressed sequence tags (ESTs) were assembled to yield 1510 unique sequences with an average length of 712 bp. The majority (79.4%) of the unique sequences were assigned to the putative functions based on the BLAST searches against the public databases. The functions of these unique sequences covered a broad set of molecular functions, biological processes and biochemical pathways according to GO and KEGG assignments. The transcripts involved in the secondary metabolite biosynthesis of alkaloids, terpenoids and flavone/flavonoids, such as cytochrome P450, lysine decarboxylase (LDC), flavanone 3-hydroxylase, squalene synthetase and 2-oxoglutarate 3-dioxygenase, were well represented by 34 unique sequences in this EST dataset. The corresponding peptide sequence of the LDC contained the Pfam 03641 domain and was annotated as a putative LDC. The unique sequences encoding transcription factors, phytohormone biosynthetic enzymes and signaling components were also found in this EST collection. In addition, a total of 501 potential SSR-motif microsatellite loci were identified from the 393 H. serrata leaf unique sequences. This set of non-redundant ESTs and the molecular markers obtained in this study will establish valuable resources for a wide range of applications including gene discovery and identification, genetic mapping and analysis of genetic diversity, cultivar identification and marker-assisted selections in this important medicinal plant.