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1.
Zhongguo Zhong Yao Za Zhi ; 44(13): 2777-2784, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31359690

RESUMO

According to the data of Pinellia ternate transcriptome,two calmodulin genes were cloned and named as Pt Ca M1 and PtCa M2 respectively. The results of bioinformatics analysis showed that Pt Ca Ms genes contained a 450 bp open reading frame,encoding149 amino acids.The identity of the coding sequences was 80%,and the identity of amino acids sequence was 91%. Pt Ca Ms genes contained EF-hand structure domain,belonging to the Ca M families. The Real-time PCR analysed the expression patterns of Pt Ca Ms in different tissues and different treatments. RESULTS:: showed that Pt Ca M1 and Pt Ca M2 gene were the highest expression level in tuber. Under Ca Cl2 treatment,the expressions of Pt Ca Ms were significantly higher than the control. Under EGTA,La Cl3 and TFP treatments,the expression level of Pt Ca Ms decreased gradually. In this study,the Pt Ca Ms gene were successfully cloned from P. ternate,which laid a foundation for the functional characteristic of Pt Ca Ms gene and the synthesis of alkaloids from P. ternata for further study.


Assuntos
Calmodulina/genética , Pinellia/genética , Clonagem Molecular , Genes de Plantas , Tubérculos/genética
2.
Nat Prod Res ; : 1-7, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30873860

RESUMO

One new neolignan glycoside, dolomiside A (1), together with 11 known phenylpropanoid glycosides were isolated from Dolomiaea souliei (Franch.) Shih. The structures of these isolates were determined by UV, CD, HR-ESI-TOFMS, 1D and 2D NMR analysis.

3.
Mar Drugs ; 16(9)2018 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-30208577

RESUMO

Seven long-chain amides, including five previously undescribed bacillamidins A⁻E (1⁻5) and two previously reported synthetic analogs, bacillamidins F (6) and G (7), were isolated from extracts of the marine-derived Bacillus pumilus strain RJA1515. The structures of the new compounds were established by extensive analysis of 1D and 2D nuclear magnetic resonance (NMR) data as well as high resolution mass spectrometry (HRMS), and the absolute configurations of the stereogenic carbons of 1⁻4 were established by comparison of the calculated and the experimental electronic circular dichroism (ECD) spectra. The cytotoxic and antimicrobial activities of 1⁻7 were evaluated.


Assuntos
Amidas/farmacologia , Anti-Infecciosos/farmacologia , Antineoplásicos/farmacologia , Organismos Aquáticos/química , Bacillus pumilus/química , Amidas/química , Amidas/isolamento & purificação , Anti-Infecciosos/química , Anti-Infecciosos/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Bactérias/efeitos dos fármacos , Linhagem Celular Tumoral , Dicroísmo Circular , Humanos , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Estrutura Molecular
4.
Zhongguo Zhong Yao Za Zhi ; 43(11): 2264-2260, 2018 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-29945376

RESUMO

Artemisinin is a preferred medicine in the treatment of malaria. In this study, AaCMK, a key gene involved in the upstream pathway of artemisinin biosynthesis, was cloned and characterized from Artemisia annua for the first time. The full-length cDNA of AaCMK was 1 462 bp and contained an ORF of 1 197 bp that encoded a 399-anomo-acid polypeptide. Tissue expression pattern analysis showed that AaCMK was expressed in leaves, flowers, roots and stems, but with higher expression level in glandular secretory trichomes. In addition, the expression of AaCMK was markedly increased after MeJA treatment. Subcellular localization showed that the protein encoded by AaCMK was localized in chloroplast. Overexpression of AaCMK in Arabidopsis increased the contents of chlorophyll a, chlorophyll b and carotenoids. These results suggest that AaCMK plays an important role in the biosynthesis of terpenoids in A. annua and this research provids a candidate gene that could be used for engineering the artemisinin biosynthesis.


Assuntos
Artemisia annua/genética , Proteínas de Plantas/genética , Artemisia annua/enzimologia , Artemisininas , Clorofila A , Clonagem Molecular
5.
Zhongguo Zhong Yao Za Zhi ; 43(1): 72-78, 2018 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-29552814

RESUMO

Hyoscyamine and scopolamine are two main alkaloids in Atropa belladonna with great medicinal value. In this paper, the contents of hyoscyamine and scopolamine, the upstream products in alkaloid synthesis, and the expression levels of key enzyme genes PMT, TRⅠ and H6H in secondary metabolism of A. belladonna seedlings were measured to clarify the mechanism of nitrogen forms regulating alkaloids synthesis.The results showed that the 50/50 (NH⁺4/NO⁻3) treatment was more favorable for the accumulation of alkaloids and the conversion of hyoscyamine to scopolamine. The content of putrescine was almost consistent with the change of key enzymes activities in the synthesis of putrescine, they both increased with the rise of ammonium ratio, reaching the highest at 75/25 (NH⁺4/NO⁻3). The detection of signaling molecule nitric oxide (NO) showed that the NO concentration decreased with the decrease of nitrate proportion. Further detection of gene expression levels of PMT, TRⅠ and H6H in TAs synthesis pathway showed that a certain amount of ammonium promoted the expression of PMT and H6H in roots. When the ratio of ammonium to nitrate was 50/50, PMT, TRⅠ and H6H in leaves and roots had higher expression levels. It can be speculated that the regulation of the formation of hyoscyamine to scopolamine by nitrogen forms mainly through affecting the expression of key enzyme genes. 50/50 (NH⁺4/NO⁻3) treatment increased the gene expression of TRⅠ in both leaves and roots as well as PMT and H6H in roots, promoting the synthesis of putrescine to hyoscyamine and the conversion of hyoscyamine to scopolamine.


Assuntos
Atropa belladonna/enzimologia , Hiosciamina/biossíntese , Nitrogênio/metabolismo , Escopolamina/metabolismo , Atropa belladonna/genética , Regulação da Expressão Gênica de Plantas , Oxigenases de Função Mista
6.
Nat Prod Res ; 32(12): 1432-1435, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28637366

RESUMO

The aerial parts of Epilobium plants are widely used as folk medicine and food around the world. The present study was aimed to investigate the antioxidant activities and active chemical constituents from Epilobium angustifolium L. The results revealed that the EtOAc extract, rich in phenolic compounds and flavonoids (16.81 ± 0.67 g GAE/100 g extract and 4.95 ± 0.21 g QE/100 g extract, respectively), possessed significantly antioxidant activities in reducing power, DPPH radical scavenging activity, ABTS radical scavenging activity and highly in inhibiting lipid peroxidation activity. Simultaneously, active fractions F to H from EtOAc extracts showing potent in vitro antioxidant activities also contained high content of total phenolic and flavonoid. Twenty-eight compounds were identified as phenolic compounds and flavonoids by LC-MS/MS. The results illustrate that the E. angustifolium L., which is rich in phenolics, could be used as a natural resource of antioxidant ingredient.


Assuntos
Antioxidantes/farmacologia , Epilobium/química , Fenóis/análise , Extratos Vegetais/farmacologia , Antioxidantes/química , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/análise , Flavonoides/química , Depuradores de Radicais Livres/química , Depuradores de Radicais Livres/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fenóis/química , Componentes Aéreos da Planta/química , Extratos Vegetais/análise , Extratos Vegetais/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo
7.
Zhongguo Zhong Yao Za Zhi ; 42(16): 3143-3145, 2017 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-29171233

RESUMO

A new napthalenone, rumexone A (1), was isolated from the roots of Rumex nepalensis. The structure of 1 was elucidated by extensive spectroscopic analyses, including 1D and 2D NMR spectra and MS data. Its cytotoxic effect was evaluated using four clinically relevant human cancer cell lines, gastric carcinoma SGC7901, breast carcinoma MDA-MB-231, lung carcinoma A549, and hepatocellular carcinoma HepG2.


Assuntos
Naftalenos/isolamento & purificação , Raízes de Plantas/química , Rumex/química , Linhagem Celular Tumoral , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Naftalenos/farmacologia , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia
8.
Oncotarget ; 8(43): 75195-75205, 2017 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-29088857

RESUMO

BACKGROUND: Inflammation and nutrition are two main causes contributing to progression of gastric cancer (GC), and inflammatory biomarker may be presented as its valuable prognostic factor. Thus, this study was carried out to investigate the prognostic significance of preoperative circulating albumin/fibrinogen ratio (AFR), fibrinogen/pre-Albumin ratio (FPR), fibrinogen (Fib), albumin (Alb) and pre-Albumin (pAlb) in surgical GC. MATERIALS AND METHODS: Three hundred and sixty surgical stage II and III GC patients from June 2011 to December 2013 were enrolled in this retrospective study. X-tile software, Kaplan-Meier curve and Cox regression model were used to evaluate the prognostic role of them. A predictive nomogram was established to predict prognosis of overall survival (OS), and its accuracy was assessed by concordance index (c-index). RESULTS: Decreased Alb, pAlb, AFR and elevated FPR were significantly associated with shorter OS. FPR was identified as the most effective prognostic factor to predict 3-year's OS by time-dependent ROC analysis. A long survival was observed in patients with low level of FPR and the prognosis of stage III FPR-low GC patients undergoing chemotherapy was significantly superior to the patients without the treatment (P=0.002). However, no difference of survival was examined in stage II subgroups stratified by FPR and high FRP of stage III patients with or not the treatment of chemotherapy. C-index of nomogram containing FPR (c-index=0.756) was high in comparison with the nomogram without FPR (c-index =0.748). CONCLUSION: Preoperative FPR might be a feasible prognostic biomarker in surgical stage II and III GC and it could precisely distinguish stage III patients who appeared to obviously benefit from adjuvant chemotherapy. Meanwhile established nomogram based on clinical parameters and FPR could improve its predictive efficacy.

9.
J Asian Nat Prod Res ; 19(12): 1172-1176, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28366016

RESUMO

Two new long-chain unsaturated compounds, (2E, 6E)-10-methoxy-3,7-dimethyl-10-oxodeca-2,6-dienoic acid (1) and (2E, 6E)-3,7,11-trimethyldodeca-2,6-dienedioic acid (2), together with seven known compounds were isolated from a marine-derived Streptomyces sp. Their structures were determined by spectroscopic methods, including 2D NMR techniques. Compounds 1 and 2 were investigated for their antibacterial activities.


Assuntos
Antibacterianos/isolamento & purificação , Ácidos Graxos Insaturados/isolamento & purificação , Streptomyces/química , Antibacterianos/química , Antibacterianos/farmacologia , Ácidos Graxos Insaturados/química , Ácidos Graxos Insaturados/farmacologia , Biologia Marinha , Testes de Sensibilidade Microbiana , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular
10.
J Ethnopharmacol ; 198: 139-147, 2017 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-28065777

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The ripe seeds of Herpetospermum caudigerum have been used in Tibetan folk medicine for treatment of bile or liver diseases including jaundice, hepatitis, intumescences or inflammation. Previously reports suggested that the seed oil and some lignans from H. caudigerum exhibited protective effects against carbon tetrachloride (CCl4)-induced hepatic damage in rats, which may be related to their free radical scavenging effect. However, the protective effect of H. caudigerum against cholestasis is still not revealed. The aim of the present study was to investigate the pharmacological effect and the chemical constituents of the petroleum ether extract (PEE) derived from H. caudigerum against α-naphthylisothiocyanate (ANIT)-induced acute cholestasis in rats. MATERIALS AND METHODS: Male cholestatic Sprague-Dawley (SD) rats induced by ANIT (60mg/kg) were orally administered with PEE (350, 700 and 1400mg/kg). Levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-Glutamyl transpeptidase (γ-GTP), total bilirubin (TBIL), direct bilirubin (DBIL) and total bile acid (TBA), as well as bile flow, and histopathological assay were evaluated. Hepatic malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione S-transferase (GST), and nitric monoxide (NO) in liver were measured to explore the possible protective mechanisms. Phytochemical analysis of PEE was performed by gas chromatography-mass spectrometer (GC-MS). RESULTS: PEE have exhibited significant and dose-dependent protective effect on ANIT-induced liver injury by reduce the increases in serum levels of ALT, AST, ALP, γ-GTP, TBIL, DBIL and TBA, restore the bile flow in cholestatic rats, and reduce the severity of the pathological tissue damage induced by ANIT. Hepatic MDA, MPO and NO contents in liver tissue were reduced, while SOD and GST activities were elevated in liver tissue. 49 compounds were detected and 39 of them were identified by GC-MS analysis, in which long-chain fatty acids were the main constituents. CONCLUSIONS: PEE exhibited a dose-dependently protective effect on ANIT-induced liver injury in cholestatic rats with the potential mechanism of attenuated oxidative stress in the liver tissue, and the possible active compounds were long-chain fatty acids.


Assuntos
1-Naftilisotiocianato/toxicidade , Colestase/tratamento farmacológico , Cucurbitaceae , Medicina Tradicional Tibetana , Extratos Vegetais/uso terapêutico , Doença Aguda , Animais , Colestase/induzido quimicamente , Colestase/metabolismo , Cucurbitaceae/química , Relação Dose-Resposta a Droga , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/análise , Ratos , Ratos Sprague-Dawley
11.
Yao Xue Xue Bao ; 52(1): 172-9, 2017 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-29911833

RESUMO

Tropane alkaloids are anticholinergic drugs widely used clinically. Biosynthesis of tropane alkaloids in planta involves a step of transamination of phenylalanine. Based on the sequenced transcriptomes of lateral roots and leaves of Hyoscyamus niger, we found three annotated aromatic amino acid aminotransferases, which were respectively named HnArAT1, HnArAT2 and HnArAT3. Sequence analysis showed that HnArAT3 had highest similarity with the reported Atropa belladonna Ab Ar AT4, which was involved in tropane alkaloid(TA) to provide the precursor of the phenyllactic acid moiety. Tissue expression pattern analysis indicated that HnArAT3 was specifically expressed in lateral roots, where is the organ synthesizing tropane alkaloids. Then, method of virus induced gene silencing (VIGS) was used to characterize the function of HnArAT3 in H. niger. Gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had lower expression levels of HnArAT3 than the non-transgenic control, and HPLC analysis of alkaloids demonstrated significant decrease in the contents of hyoscyamine, anisodamine and scopolamine in planta. These results suggested that HnArAT3 was involved in the phenyllactic acid branch of TA biosynthetic pathway. Molecular cloning and functional identification of HnArAT3 laid the foundation for further understanding of TA biosynthesis and metabolic regulation, and also provided a new candidate gene for engineering biosynthetic pathway of tropane alkaloids.


Assuntos
Alcaloides/biossíntese , Hyoscyamus/genética , Proteínas de Plantas/genética , Transaminases/genética , Tropanos/metabolismo , Atropa belladonna , Vias Biossintéticas , Antagonistas Colinérgicos , Clonagem Molecular , Hiosciamina , Hyoscyamus/enzimologia , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Escopolamina , Alcaloides de Solanáceas
12.
J Asian Nat Prod Res ; 19(2): 157-163, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27285511

RESUMO

Phytochemical investigation on the stems of Kadsura renchangiana led to the isolation of two new sesquiterpenoids, renchangianins F and G (1 and 2). Their structures were elucidated by spectroscopic methods, including 2D NMR techniques. The in vitro cytotoxic activities of the isolates were studied against HepG2, A549, and LN229 cancer cell lines.


Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Kadsura/química , Sesquiterpenos/isolamento & purificação , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Células Hep G2 , Humanos , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Caules de Planta/química , Sesquiterpenos/química , Sesquiterpenos/farmacologia
13.
Nat Prod Res ; 31(16): 1855-1860, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27871187

RESUMO

One new triterpenoid (1) and 13 known compounds (2-14) were isolated from Schisandra pubescens stems. The structure of the new compound was established on the basis of 1D/2D NMR and HRESIMS spectroscopic analyses. The isolated compounds were evaluated for their hepatoprotective activities against D-GalN-induced cell injury in QSG7701 cells. Compounds 1, 13 and 14 at 10 µM showed hepatoprotective activities, with survival rates of 60.5, 50.4 and 48.9%, respectively.


Assuntos
Lignanas/farmacologia , Substâncias Protetoras/química , Substâncias Protetoras/farmacologia , Schisandra/química , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos/métodos , Hepatócitos/efeitos dos fármacos , Humanos , Lignanas/química , Lignanas/isolamento & purificação , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Caules de Planta/química , Triterpenos/química , Triterpenos/farmacologia
14.
Nat Prod Res ; 31(9): 1034-1041, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27931116

RESUMO

One new compound, himalain A (1), together with 12 known compounds were isolated from Mirabilis himalaica. Their structures were determined by spectroscopic methods, including 2D NMR techniques, and the absolute configuration of the 1,2-diol moiety in 1 was defined through Riguera's method. All isolated compounds were tested for their cytotoxic and antibacterial activities.


Assuntos
Mirabilis/química , Éteres Fenílicos/isolamento & purificação , Antibacterianos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Humanos , Éteres Fenílicos/química , Éteres Fenílicos/farmacologia
15.
J Asian Nat Prod Res ; 18(11): 1108-14, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27248222

RESUMO

One new cyanoside, rhobupcyanoside B (1), together with 7 known ones, was isolated from the 70% ethanol extract of the roots and rhizomes of Rhodiola bupleuroides. Their structures were determined by spectroscopic methods, including 2D NMR techniques. Compound 1 was evaluated for its inhibitory activity against α-glucosidase with IC50 value of 278.28 ± 0.55 µM by comparing with the positive control (acarbose) at 210.40 ± 0.32 µM.


Assuntos
Glicosídeos/isolamento & purificação , Rhodiola/química , alfa-Glucosidases/efeitos dos fármacos , Inibidores de Glicosídeo Hidrolases/química , Glicosídeos/química , Glicosídeos/farmacologia , Concentração Inibidora 50 , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Raízes de Plantas/química
16.
Yao Xue Xue Bao ; 51(5): 770-4, 2016 05.
Artigo em Chinês | MEDLINE | ID: mdl-29874024

RESUMO

The chemical constituents of Herpetospermum caudigerum were investigated using chromatographic methods, including silica gel column chromatography, Sephadex LH-20 and semi-preparative HPLC. Four compounds were isolated and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2,11-dimethoxy-3,9-dihydroxy-7 H-dibenzo[c,e]oxepin-5-one (1),7,8'-didehydroherpetotriol(2), herpetotrio l(3) and kaempferitrin (4). Among those, compound 1 is one new 7H-dibenzo[c,e]oxepin-5-one, named as herpetolide C.


Assuntos
Cucurbitaceae/química , Oxepinas/isolamento & purificação , Cromatografia Líquida de Alta Pressão
17.
Yao Xue Xue Bao ; 51(8): 1334-9, 2016 08.
Artigo em Chinês | MEDLINE | ID: mdl-29906047

RESUMO

The plastidial methylerythritol phosphate(MEP) pathway provides 5-carbon precursors to the biosynthesis of isoprenoid (including artemisinin). 2-C-Methyl-D-erythritol-4-phosphate cytidylyltransferase (MCT) is the third enzyme of the MEP pathway, which catalyzes 2-C-methyl-D-erythritol-4-phosphate to form 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol. The full-length MCT cDNA sequence (AaMCT) was cloned and characterized for the first time from Artemisia annua L. Analysis of tissue expression pattern revealed that AaMCT was highly expressed in glandular secretory trichome and poorly expressed in leaf, flower, root and stem. AaMCT was found to be a methyl jasmonate (Me JA)-induced genes, the expression of AaMCT was significantly increased after MeJA treatment. Subcellular localization indicated that the GFP protein fused with AaMCT was targeted specifically in chloroplasts. The transgenic plants of Arabidopsis thaliana with AaMCT overexpression exhibited a significantly increase in the content of chlorophyll a, chlorophyll b and carotenoids, demonstrating that AaMCT kinase plays an influential role in isoprenoid biosynthesis.


Assuntos
Artemisia annua/enzimologia , Nucleotidiltransferases/genética , Proteínas de Plantas/genética , Acetatos , Arabidopsis , Artemisia annua/genética , Artemisininas , Carotenoides/análise , Clorofila/análise , Clorofila A , Clonagem Molecular , Ciclopentanos , DNA Complementar , Regulação da Expressão Gênica de Plantas , Oxilipinas , Plantas Geneticamente Modificadas
18.
Yao Xue Xue Bao ; 51(11): 1791-8, 2016 11.
Artigo em Chinês | MEDLINE | ID: mdl-29908529

RESUMO

Artemisinin is the first choice for malaria treatment. The plastidial MEP pathway provides 5-carbon precursors (IPP and its isomer DMAPP) for the biosynthesis of isoprenoid (including artemisinin). Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) is the last enzyme involved in the MEP pathway, which catalyzes HMBPP to form IPP and DMAPP. In this study, we isolated the full-length cDNA of HDR from Artemisia annua L. (AaHDR2) and performed functional analysis. According to gene expression analysis of AaHDR2 (GenBank: KX058541) and AaHDR1 reported ever (GenBank: ADC84348.1) by qPCR, we found that AaHDR1 and AaHDR2 had much higher expression level in trichomes than that in roots, stems, leaves and flowers. AaHDR2 had much higher expression level in flowers than that in leaves. Further, the plant hormones such as Me JA and ABA respectively up-regulated the expression level of AaHDR1 and AaHDR2 significantly, but GA3 up-regulated the expression level of AaHDR2 only. The gene expression analysis of AaHDR1 and AaHDR2 showed that AaHDR2 had a greater contribution than AaHDR1 to isoprenoid biosynthesis(including artemisinin). We used AaHDR2 for the following experiments. Bioinformatic analysis indicated that AaHDR2 belonged to the HDR family and the functional complementation assay showed that AaHDR2 did have the enzymatic function of HDR, using E. coli mutant MG1655(ara)<>HDR as host cell. The subcellular localization assay showed that AaHDR2 fused with GFP at its N-terminal specifically targeted in chloroplasts. Finally, AaHDR2 was overexpressed in Arabidopsis thaliana. The AaHDR2-overexpressing plants produced the isoprenoids including chlorophyll a, chlorophyll b and carotenoids at significantly higher levels than the wild-type Arabidopsis plants. In summary, AaHDR2 might be a candidate gene for genetic improvement of the isoprenoid biosynthesis.


Assuntos
Artemisia annua/genética , Oxirredutases/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Arabidopsis , Artemisia annua/enzimologia , Carotenoides , Clorofila , Clorofila A , Cloroplastos , Clonagem Molecular , DNA Complementar , Escherichia coli , Reguladores de Crescimento de Planta , Terpenos/metabolismo
19.
Yao Xue Xue Bao ; 51(12): 1913-9, 2016 12.
Artigo em Chinês | MEDLINE | ID: mdl-29923697

RESUMO

Atropa belladonna L. is the commercial plant material for production of tropane alkaloids, including hyoscyamine and scopolamine. The wild-type Atropa belladonna is characterized by the hyoscyamine-rich chemotype, in which the hyoscyamine content is much higher than the scopolamine content. It is the common goal for the pharmaceutical industry to increase the content of scopolamine in A. belladonna. Based on the T0 progeny of transgenic A. belladonna with NtPMT and HnH6H overexpression, T1 progeny of transgenic A. belladonna were obtained through self-pollination and used in a field trial. The 461 bp fragment of NtPMT and the 1 077 bpHnH6 H were simultaneously expressed from T1 progeny of transgenic A. belladonna, but were not obtained from the wild-type A. belladonna. At the transcription level, the expression of NtPMT and HnH6H were detected in T1 progeny of transgenic A. belladonna, but were not detected in the wild-type plants. Further, the alkaloids were analyzed by HPLC. In the stems and leaves of T1 progeny of transgenic A. belladonna, hyoscyamine was not detected and scopolamine was detected at very high levels; in the stems and leaves of wild-type A. belladonna, hyoscyamine was detected at much higher levels. In the leaves of T1 progeny of transgenic A. belladonna, the content of scopolamine was 15-36 folds higher than that of wild- type leaves; in the stems of T1 progeny of transgenic A. belladonna, the scopolamine content was 37-108 folds higher than that of wild-type stems. In conclusion, overexpression of NtPMT and HnH6H greatly enhanced conversion of hyoscyamine into high-value scopolamine and improved the commercial value of A. belladonna.


Assuntos
Atropa belladonna/química , Atropa belladonna/genética , Hiosciamina/análise , Plantas Geneticamente Modificadas/química , Escopolamina/análise , Alcaloides , Antagonistas Colinérgicos , Regulação da Expressão Gênica de Plantas , Oxigenases de Função Mista , Tropanos
20.
Yao Xue Xue Bao ; 51(3): 408-10, 2016 03.
Artigo em Chinês | MEDLINE | ID: mdl-29859021

RESUMO

A new isocoumarin, chamerilactone A(1) was isolated from the ethanol extract of Chamerion angustifolium with normal phase silica column chromatography, Sephadex LH-20, MCI CHP-20 and semi- preparative HPLC methods. Its structure and stereochemistry were elucidated by spectroscopic methods, including 2D-NMR and optical rotation techniques.


Assuntos
Isocumarinas/isolamento & purificação , Onagraceae/química , Cromatografia Líquida de Alta Pressão , Estrutura Molecular
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