RESUMEN
Isopentenyl diphosphate isomerase (IDI), a key enzyme in the biosynthetic pathway of diterpenoid alkaloids (DAs), plays an essential regulatory role in the synthesis and accumulation of DAs. In this study, the coding sequence (CDS) of AcIDI1 was isolated from the mother roots of Aconitum carmichaelii Debx. (GeneBank accession number OR915879). Bioinformatics analysis showed that the CDS of AcIDI1 was 894 bp, encoding a protein with 297 amino acids and the putative protein localized in the chloroplast. AcIDI1 exhibited significant homology with sequences encoding IDI in other species, and was most closely related to Aconitum vilmorinianum. Furthermore, the fusion protein has been successfully expressed in Escherichia coli (E. coli), providing a basis for future functional studies of AcIDI1. The expression pattern of AcIDI1 was analyzed by real-time quantitative PCR (qPCR), which demonstrates that AcIDI1 is a tissue-specific gene in the roots of A. carmichaelii and exhibits high expression in both daughter and mother roots. By comparing the expression levels of AcIDI1 in three tissues of the roots of A. carmichaelii at different growth stages, we propose that the mother roots (MRs) are the centers of resources allocation. The roots of A. carmichaelii continuously absorb the energy from external environment, while resources transfer behavior from MRs to both daughter roots (DRs) and axillary buds (ABs) occurs as the plant grows. This study establishes a foundation for applying the IDI gene to regulate the biosynthesis and accumulation of DAs in A. carmichaelii.
Asunto(s)
Aconitum , Alcaloides , Diterpenos , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Raíces de Plantas , Aconitum/genética , Aconitum/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/genética , Diterpenos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Alcaloides/metabolismo , Alcaloides/biosíntesis , Filogenia , Escherichia coli/genética , Escherichia coli/metabolismoRESUMEN
In this study, we conducted an assembly and analysis of the organelle genomes of Aconitum carmichaelii. Our investigation encompassed the examination of organelle genome structures, gene transfer events, and the environmental selection pressures affecting A. carmichaelii. The results revealed distinct evolutionary patterns in the organelle genomes of A. carmichaelii. Especially, the plastome exhibited a more conserved structure but a higher nucleotide substitution rate (NSR), while the mitogenome displayed a more complex structure with a slower NSR. Through homology analysis, we identified several instances of unidirectional protein-coding genes (PCGs) transferring from the plastome to the mitogenome. However, we did not observe any events which genes moved from the mitogenome to the plastome. Additionally, we observed multiple transposable element (TE) fragments in the organelle genomes, with both organelles showing different preferences for the type of nuclear TE insertion. Divergence time estimation suggested that rapid differentiation occurred in Aconitum species approximately 7.96 million years ago (Mya). This divergence might be associated with the reduction in CO2 levels and the significant uplift of the Qinghai-Tibet Plateau (QTP) during the late Miocene. Selection pressure analysis indicated that the dN/dS values of both organelles were less than 1, suggested that organelle PCGs were subject to purification selection. However, we did not detect any positively selected genes (PSGs) in Subg. Aconitum and Subg. Lycoctonum. This observation further supports the idea that stronger negative selection pressure on organelle genes in Aconitum results in a more conserved amino acid sequence. In conclusion, this study contributes to a deeper understanding of organelle evolution in Aconitum species and provides a foundation for future research on the genetic mechanisms underlying the structure and function of the Aconitum plastome and mitogenome.
Asunto(s)
Aconitum , Filogenia , Aconitum/genética , Aconitum/química , Aconitum/metabolismo , Orgánulos/genética , TibetRESUMEN
Diterpenoid alkaloids (DAs) have been often utilized in clinical practice due to their analgesic and anti-inflammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown. In this study, we successfully assembled a high-quality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum (A. vilmorinianum) (5.76 Gb). An A. vilmorinianum-specific whole-genome duplication event was discovered using comparative genomic analysis, which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A. vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.
Asunto(s)
Aconitum , Alcaloides , Diterpenos , Aconitum/genética , Aconitum/metabolismo , Multiómica , Diterpenos/metabolismo , Alcaloides/metabolismo , Transcriptoma/genética , Raíces de PlantasRESUMEN
Diterpene alkaloids (DAs) are characteristic compounds in Aconitum, which are classified into four skeletal types: C18, C19, C20, and bisditerpenoid alkaloids. C20-DAs are thought to be the precursor of the other types. Their biosynthetic pathway, however, is largely unclear. Herein, we combine metabolomics and transcriptomics to unveil the methyl jasmonate (MJ) inducible biosynthesis of DAs in the sterile seedling of A. gymnandrum, the only species in the Subgenus Gymnaconitum (Stapf) Rapaics. Target metabolomics based on root and aerial portions identified 51 C19-DAs and 15 C20-DAs, with 40 inducible compounds. The highest content of C20-DA atisine was selected for further network analysis. PacBio Isoform sequencing integrated with RNA sequencing not only provided the full-length transcriptome but also their response to induction, revealing 1994 genes that exhibited up-regulated expression. Further, 38 genes involved in terpenoid biosynthesis were identified, including 7 diterpene synthases. In addition to the expected function of the four diterpene synthases, AgCPS5 was identified to be a new ent-8,13-CPP synthase in Aconitum and could also combine with AgKSL1 to form the C20-DAs precursor ent-atiserene. Combined with multiple network analyses, six CYP450 and seven 2-ODD genes predicted to be involved in the biosynthesis of atisine were also identified. This study not only sheds light on diterpene synthase evolution in Aconitum but also provides a rich dataset of full-length transcriptomes, systemic metabolomes, and gene expression profiles, setting the groundwork for further investigation of the C20-DAs biosynthesis pathway.
Asunto(s)
Aconitum , Alcaloides , Diterpenos , Aconitum/genética , Aconitum/metabolismo , Transcriptoma , Alcaloides/metabolismo , Diterpenos/metabolismo , Vías Biosintéticas/genéticaRESUMEN
Aconitum, as "the first drug of choice for invigorating Yang and saving lives", has been widely used for the treatment of heart failure. However, toxic components of Aconitum can easily lead to serious arrhythmia, even death (Y. CT., 2009; Zhang XM., 2018). In this study, a High Performance Liquid Chromatography (HPLC) method for the determination of aconitine (AC), mesaconitine (MA) and hypaconitine (HA) was established; The effect of Glycyrrhiza on CYP3A1 / 2 mRNA expression was detected by RT-PCR; SD rats were given Aconitum and compatibility of Glycyrrhizae and Aconitum by gavage respectively, the blood concentration of toxic components were determined by LC-MS / MS; The CHF rat model was established by intraperitoneal injection of adriamycin (2.5 mg / kg), and were randomly divided into model, Aconitum, the compatibility of Glycyrrhizae and Aconitum and Captopril group, 5 mice/group. After 4 weeks of gavage, the corresponding indexes were detected by ELISA and HPLC. The results showed that Ketoconazole significantly inhibited the metabolites of AC, MA and HA; Glycyrrhiza induced CYP3A gene expression; The level of ALD in the compatibility of Glycyrrhizae and Aconitum group was significantly lower than that in Aconitum group. After intervention with the compatibility of Glycyrrhizae and Aconitum, ATP increased, ADP decreased significantly. In conclusion, we found Glycyrrhiza promoted the metabolism of toxic components of Aconitum by up regulating the expression of CYP3A, and reduced the content of BNP, Ang II and ALD, improved the energy metabolism disorder of myocardium, alleviated the development of CHF.
Asunto(s)
Aconitum , Medicamentos Herbarios Chinos , Glycyrrhiza uralensis , Insuficiencia Cardíaca , Aconitina/farmacología , Aconitum/metabolismo , Aconitum/toxicidad , Animales , Cromatografía Líquida de Alta Presión/métodos , Citocromo P-450 CYP3A/genética , Medicamentos Herbarios Chinos/farmacología , Glycyrrhiza uralensis/metabolismo , Insuficiencia Cardíaca/prevención & control , Ratones , Extractos Vegetales/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
In recent decades, herbal medicines have played more and more important roles in the healthcare system in the world because of the good efficacy. However, with the increasing use of herbal medicines, the toxicity induced by herbal medicines has become a global issue. Therefore, it is needed to investigate the mechanism behind the efficacy and toxicity of herbal medicines. In this study, using Aconiti Lateralis Radix Praeparata (Fuzi) as an example, we adopted a systems pharmacology approach to investigate the mechanism of Fuzi in treating rheumatoid arthritis and in inducing cardiac toxicity and neurotoxicity. The results showed that Fuzi has 25 bioactive compounds that act holistically on 61 targets and 27 pathways to treat rheumatoid arthritis, and modulation of inflammation state is one of the main mechanisms of Fuzi. In addition, the toxicity of Fuzi is linked to 32 compounds that act on 187 targets and 4 pathways, and the targets and pathways can directly modulate the flow of Na+, Ca2+, and K+. We also found out that non-toxic compounds such as myristic acid can act on targets of toxic compounds and therefore may influence the toxicity. The results not only reveal the efficacy and toxicity mechanism of Fuzi, but also add new concept for understanding the toxicity of herbal medicines, i.e., the compounds that are not directly toxic may influence the toxicity as well.
Asunto(s)
Aconitum/metabolismo , Artritis Reumatoide/tratamiento farmacológico , Diterpenos/farmacología , Diterpenos/toxicidad , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/toxicidad , Algoritmos , Química Farmacéutica/métodos , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Humanos , Medicina Tradicional China/métodos , Farmacología en Red/métodos , Extractos Vegetales/farmacología , Plantas Medicinales/metabolismo , Mapeo de Interacción de ProteínasRESUMEN
The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).
Asunto(s)
Aconitum , Alcaloides , Diterpenos , Acetilcolinesterasa , Aconitum/metabolismo , Espectroscopía de Resonancia Magnética , Estructura Molecular , Raíces de Plantas/metabolismoRESUMEN
Natural plants must actively allocate their limited resources for survival and reproduction. Although vegetative growth, sexual reproduction, asexual reproduction and defense are all basic processes in the life cycle of plants, the strategies used to allocate resources between these processes are poorly understood. These processes are conspicuous in naturally grown Aconitum kusnezoffii Reichb., which makes it a suitable study subject. Here, the morphology, dry matter, total organic carbon, total nitrogen and aconitum alkaloid levels of shoot, principal root (PR) and lateral roots were measured throughout the growing season. Then, transcriptome and metabolite content analyses were performed. We found that vegetative growth began first. After vegetative growth ceased, sexual development began. Flower organ development was accompanied by increased photosynthesis and the PR consumed temporarily stored resources after flower formation. Asexual propagule development initiated earlier than sexual reproduction and kept accumulating resources after that. Development was slow before flower formation, mainly manifesting as increasing length; then, after flower formation it accelerated via enhanced material transport and accumulation. Defense compounds were maintained at low levels before flowering. In particular, the turnover of defense compounds was enhanced before and after flower bud emergence, providing resources for other processes. After flower formation, defense compounds were accumulated. The pattern found herein provides a vivid example for further studies on resource allocation strategies. The exciting finding that the PR, as a more direct storage site for photosynthate, is a buffer unit for resources, and that defense compounds can be reused for other processes, suggests a need to explore potential mechanisms.
Asunto(s)
Aconitum/fisiología , Reproducción Asexuada , Aconitum/crecimiento & desarrollo , Aconitum/metabolismo , Carbono/metabolismo , Flores/crecimiento & desarrollo , Flores/metabolismo , Flores/fisiología , Nitrógeno/metabolismo , Fotosíntesis , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Raíces de Plantas/fisiología , Brotes de la Planta/crecimiento & desarrollo , Brotes de la Planta/metabolismo , Brotes de la Planta/fisiología , Reproducción/fisiología , Reproducción Asexuada/fisiología , Estaciones del AñoRESUMEN
The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).
Asunto(s)
Acetilcolinesterasa , Aconitum/metabolismo , Alcaloides , Diterpenos , Espectroscopía de Resonancia Magnética , Estructura Molecular , Raíces de Plantas/metabolismoAsunto(s)
Aconitina/análogos & derivados , Tolerancia a Medicamentos , Morfina/uso terapéutico , Fibras Nerviosas Amielínicas/metabolismo , Neuroglía/metabolismo , Proteína Quinasa C/metabolismo , Asta Dorsal de la Médula Espinal/metabolismo , Sinapsis/metabolismo , Aconitina/administración & dosificación , Aconitina/uso terapéutico , Aconitum/metabolismo , Administración Oral , Animales , Astrocitos/metabolismo , Conducta Animal , Dolor Crónico , Potenciación a Largo Plazo , Vértebras Lumbares/efectos de los fármacos , Masculino , Microglía/metabolismo , Ratas , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Regulación hacia ArribaRESUMEN
Endophytes have been recognized as a source for structurally novel and biologically active secondary metabolites. Among the host plants for endophytes, some medicinal plants that produce pharmaceuticals have been reported to carry endophytes, which could also produce bioactive secondary metabolites. In this study, the medicinal plant Aconitum carmichaeli was selected as a potential source for endophytes. An endophytic microorganism, Aureobasidium pullulans AJF1, harbored in the flower of Aconitum carmichaeli, was cultured on a large scale and extracted with an organic solvent. Extensive chemical investigation of the extracts resulted in isolation of three lipid type compounds (1-3), which were identified to be (3R,5R)-3,5-dihydroxydecanoic acid (1), (3R,5R)-3-(((3R,5R)-3,5-dihydroxydecanoyl)oxy)-5-hydroxydecanoic acid (2), and (3R,5R)-3-(((3R,5R)-5-(((3R,5R)-3,5-dihydroxydecanoyl)oxy)-3-hydroxydecanoyl)oxy)-5-hydroxydecanoic acid (3) by chemical methods in combination with spectral analysis. Compounds 2 and 3 had new structures. Absolute configurations of the isolated compounds (1-3) were established using modified Mosher's method together with analysis of NMR data for their acetonide derivatives. All the isolates (1-3) were evaluated for antibiotic activities against Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and their cytotoxicities against MCF-7 cancer cells. Unfortunately, they showed low antibiotic activities and cytotoxic activities.
Asunto(s)
Ascomicetos/metabolismo , Ácidos Decanoicos/química , Ácidos Decanoicos/metabolismo , Hidroxiácidos/química , Hidroxiácidos/metabolismo , Aconitum/genética , Aconitum/metabolismo , Antibacterianos/síntesis química , Antibacterianos/química , Antibacterianos/farmacología , Ascomicetos/genética , Bacterias/efectos de los fármacos , Ácidos Decanoicos/síntesis química , Ácidos Decanoicos/farmacología , Humanos , Hidroxiácidos/síntesis química , Hidroxiácidos/farmacología , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Estructura MolecularRESUMEN
Songorine isolated from Aconitum brachypodum Diels possesses prominent activity of inhibiting G protein-coupled receptors (GPCRs) in the early screening process. In this paper, a series of Songorine derivatives were synthesized and their inhibitory activities on GPCRs were also evaluated by using the Double Antibody Sandwich ELISA (DAS-ELISA) in vitro. Among them, three derivatives (3a, 4, 7) exhibited significant inhibitory activity against GPCRs with IC50 values of 0.08-0.29â¯nM. Moreover, the structure-activity relationships (SARs) of songorine derivatives were discussed in detail. They have great potentials as novel GPCRs antagonists in the future.
Asunto(s)
Alcaloides/química , Receptores Acoplados a Proteínas G/antagonistas & inhibidores , Aconitum/química , Aconitum/metabolismo , Alcaloides/metabolismo , Concentración 50 Inhibidora , Receptores Acoplados a Proteínas G/metabolismo , Relación Estructura-ActividadRESUMEN
Trans-ferulic acid-4-ß-glucoside (C16H20O9, TFA-4ß-G) is a monomer extracted from the Chinese medicine called radix aconiti carmichaeli (Fuzi). To date, research on this substance is lacking. Here, we found that trans-ferulic acid-4-ß-glucoside effectively promoted cold acclimatization in mice via increased heat production and alleviation of oxidative stress in a cold environment. Thus, our work indicates that ferulic acid-4-ß-glucoside is a potential therapeutic candidate for prevention and treatment of cold stress injury.
Asunto(s)
Respuesta al Choque por Frío/efectos de los fármacos , Glucósidos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Termogénesis/genética , Aclimatación/efectos de los fármacos , Aconitum/metabolismo , Tejido Adiposo Pardo/efectos de los fármacos , Tejido Adiposo Pardo/fisiología , Animales , Frío/efectos adversos , Respuesta al Choque por Frío/fisiología , Ácidos Cumáricos/metabolismo , Ácidos Cumáricos/uso terapéutico , Medicamentos Herbarios Chinos/metabolismo , Medicamentos Herbarios Chinos/uso terapéutico , Glucósidos/uso terapéutico , Humanos , Ratones , Termogénesis/efectos de los fármacosRESUMEN
Aconitum carmichaelii has long been used as a traditional Chinese medicine, and its processed lateral roots are known commonly as fuzi. Aconitine-type C19-diterpenoid alkaloids accumulating in the lateral roots are some of the main toxicants of this species, yet their biosynthesis remains largely unresolved. As a first step towards understanding the biosynthesis of aconitine-type C19-diterpenoid alkaloids, we performed de novo transcriptome assembly and analysis of rootstocks and leaf tissues of Aconitum carmichaelii by next-generation sequencing. A total of 525 unigene candidates were identified as involved in the formation of C19-diterpenoid alkaloids, including those encoding enzymes in the early steps of diterpenoid alkaloids scaffold biosynthetic pathway, such as ent-copalyl diphosphate synthases, ent-kaurene synthases, kaurene oxidases, cyclases, and key aminotransferases. Furthermore, candidates responsible for decorating of diterpenoid alkaloid skeletons were discovered from transcriptome sequencing of fuzi, such as monooxygenases, methyltransferase, and BAHD acyltransferases. In addition, 645 differentially expressed genes encoding transcription factors potentially related to diterpenoid alkaloids accumulation underground were documented. Subsequent modular domain structure phylogenetics and differential expression analysis led to the identification of BAHD acyltransferases possibly involved in the formation of acetyl and benzoyl esters of diterpenoid alkaloids, associated with the acute toxicity of fuzi. The transcriptome data provide the foundation for future research into the molecular basis for aconitine-type C19-diterpenoid alkaloids biosynthesis in A. carmichaelii.
Asunto(s)
Aconitina/metabolismo , Aconitum/genética , Aconitum/metabolismo , Alcaloides/biosíntesis , Aconitina/análogos & derivados , Aconitina/química , Aconitum/química , Alcaloides/química , Conformación Molecular , TranscriptomaRESUMEN
Aconitum carmichaelii is an important medicinal herb used widely in China, Japan, India, Korea, and other Asian countries. While extensive research on the characterization of metabolic extracts of A. carmichaelii has shown accumulation of numerous bioactive metabolites including aconitine and aconitine-type diterpene alkaloids, its biosynthetic pathway remains largely unknown. Biosynthesis of these secondary metabolites is tightly controlled and mostly occurs in a tissue-specific manner; therefore, transcriptome analysis across multiple tissues is an attractive method to identify the molecular components involved for further functional characterization. In order to understand the biosynthesis of secondary metabolites, Illumina-based deep transcriptome profiling and analysis was performed for four tissues (flower, bud, leaf, and root) of A. carmichaelii, resulting in 5.5 Gbps clean RNA-seq reads assembled into 128,183 unigenes. Unigenes annotated as possible rate-determining steps of an aconitine-type biosynthetic pathway were highly expressed in the root, in accordance with previous reports describing the root as the accumulation site for these metabolites. We also identified 21 unigenes annotated as cytochrome P450s and highly expressed in roots, which represent candidate unigenes involved in the diversification of secondary metabolites. Comparative transcriptome analysis of A. carmichaelii with A. heterophyllum identified 20,232 orthogroups, representing 30,633 unigenes of A. carmichaelii, gene ontology enrichment analysis of which revealed essential biological process together with a secondary metabolic process to be highly enriched. Unigenes identified in this study are strong candidates for aconitine-type diterpene alkaloid biosynthesis, and will serve as useful resources for further validation studies.
Asunto(s)
Aconitum/genética , Alcaloides/biosíntesis , Diterpenos/metabolismo , Proteínas de Plantas/genética , Metabolismo Secundario/genética , Transcriptoma , Aconitina/química , Aconitina/aislamiento & purificación , Aconitina/metabolismo , Aconitum/clasificación , Aconitum/metabolismo , Alcaloides/química , Alcaloides/aislamiento & purificación , Diterpenos/química , Diterpenos/aislamiento & purificación , Flores/genética , Flores/metabolismo , Regulación de la Expresión Génica de las Plantas , Ontología de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Anotación de Secuencia Molecular , Filogenia , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/clasificación , Proteínas de Plantas/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Plantas MedicinalesRESUMEN
A simple and sensitive method for determination of three aconitum alkaloids and their metabolites in human plasma was developed using matrix solid-phase dispersion combined with vortex-assisted dispersive liquid-liquid microextraction and high-performance liquid chromatography with diode array detection. The plasma sample was directly purified by matrix solid-phase dispersion and the eluate obtained was concentrated and further clarified by vortex-assisted dispersive liquid-liquid microextraction. Some important parameters affecting the extraction efficiency, such as type and amount of dispersing sorbent, type and volume of elution solvent, type and volume of extraction solvent, salt concentration as well as sample solution pH, were investigated in detail. Under optimal conditions, the proposed method has good repeatability and reproducibility with intraday and interday relative standard deviations lower than 5.44 and 5.75%, respectively. The recoveries of the aconitum alkaloids ranged from 73.81 to 101.82%, and the detection limits were achieved within the range of 1.6-2.1 ng/mL. The proposed method offered the advantages of good applicability, sensitivity, simplicity, and feasibility, which makes it suitable for the determination of trace amounts of aconitum alkaloids in human plasma samples.
Asunto(s)
Aconitum/química , Alcaloides/sangre , Microextracción en Fase Líquida , Extracción en Fase Sólida , Aconitum/metabolismo , Alcaloides/metabolismo , Cromatografía Líquida de Alta Presión , HumanosRESUMEN
The tuberous roots of Aconitum carmichaeli are largely used in traditional Chinese medicine and widely grown in Jiangyou, Sichuan, China. During the growth process, this medicinal plant releases a large amount of allelochemicals into soil, which retard the growth and development of near and late crops. Therefore, a pure culture experiment was thus carried out by seed soaking to study the allelopathic effects of extracts from tuberous roots of A. carmichaeli (ETR) on the seed germination and young seedling growth of Lolium perenne, Trifolium repens, and Medicago sativa, the late pasture grasses after cultivation of A. carmichaeli. The results showed that three pasture grasses varied significantly in seed germination and young seedling growth in response to ETR concentrations. Seed germination of M. sativa was stimulated by low ERT concentration (0.01 x g(-1)), while all of pasture grass seeds germinated poorly in solution with 1.00 g x L(-1). Seed soaking with 1.00 g x L(-1) also inhibited significantly the growth of pasture young seedlings, with M. sativa showing the highest seedling height reduction of 42.05% in seeding height, followed by T. repens (40.21%) and L. perenne with about 11%. Cultivation of L. perenne could thus be beneficial to increase whole land productivity in A. carmichaeli-pasture grass cropping systems. In addition, hydrolysis of protein, starch, and inositol phosphates was blocked and free amino acids, soluble sugars and phosphorus were decreased in seeds by seed soaking with ETR, which could be one of the reason for the inhibition of seed germination. There was a significant reduction in root vigor, nitrate reductase, and chlorophyll after the seed treatment with ETR, indicating the suppression of nutrient uptake, nitrate assimilation, and photosynthesis by allelopathic chemicals in ETR, which could lead to the slow growth rate of pasture grass seedlings.
Asunto(s)
Aconitum/química , Alelopatía , Feromonas/farmacología , Extractos Vegetales/farmacología , Raíces de Plantas/química , Poaceae/efectos de los fármacos , Aconitum/metabolismo , China , Feromonas/metabolismo , Extractos Vegetales/metabolismo , Raíces de Plantas/metabolismo , Poaceae/crecimiento & desarrolloRESUMEN
To authenticate Ayurvedic medicinal plants Ativisha (Aconitum heterophyllum) and Musta (Cyperus rotundus) at the raw drug source and in prepared herbal products, nrDNA ITS sequence based SCAR markers were designed and validated spp.-specific SCAR primers gave amplicon of 415 bp and 134 bp, respectively, in authentic species. The SCAR primers (Cyr-FP and Cyr-RP) could identify tissue sample containing 750 µg to 4.76 mg/100mg of Musta in complex mixtures of DNA extracted from commercial herbal drugs. Ativisha could not be identified through SCAR markers suggesting that authentic species may not been used to prepare herbal drugs despite its being labelled as one of the ingredients in formulations. Analysis of individual tubers of Ativisha and Musta assures the presence of admixtures in raw drug trade of Ativisha, indicates the need to monitor the basic raw material supply and concludes, supplying plant materials through cultivation to manufacturing industries can minimize the risks of adulteration.
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Aconitum/metabolismo , Núcleo Celular/metabolismo , Cyperus/metabolismo , ADN de Plantas/metabolismo , ADN Espaciador Ribosómico/metabolismo , Preparaciones de Plantas/química , Tubérculos de la Planta/metabolismo , Aconitum/química , Aconitum/crecimiento & desarrollo , Secuencia de Bases , Biomarcadores/metabolismo , Cyperus/química , Cyperus/crecimiento & desarrollo , ADN de Plantas/química , ADN Espaciador Ribosómico/química , Suplementos Dietéticos/análisis , Contaminación de Medicamentos/prevención & control , Contaminación de Alimentos/prevención & control , Inspección de Alimentos/métodos , India , Medicina Ayurvédica , Datos de Secuencia Molecular , Farmacognosia/métodos , Tubérculos de la Planta/química , Tubérculos de la Planta/crecimiento & desarrollo , Control de Calidad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Especificidad de la EspecieRESUMEN
Aconitum heterophyllum is a high altitude medicinal plant that has become endangered due to overexploitation for their aconitins. The most effective, conventional propagation method for any plant species is by seed. However, in Aconitum seed germination is erratic, and seedling survival is low. In the present study results have been discussed on the possible implication of ethanol treatment on removal of barriers on radical emergence in terms of protein changes. Eighty seven percent of seed germination was achieved in Aconitum with ethanol treatment. Comparative 2-DE analysis of ethanol treated and untreated seed protein profiles in Phase II of germination revealed 40 differentially expressed proteins. Twenty-seven out of 40 proteins were induced, 5 were increased and 8 were repressed. Mass spectrometry and subsequent identification confirmed that these proteins were involved in metabolism, DNA regulation, stress tolerance and plasmamembrane/cell wall biosynthesis/extension processes. These protein changes might be responsible for physiological and physical changes, respectively, resulted in increase in germination percentage. Further, characterization of these proteins will be of great help in understanding the molecular mechanism lying behind enhanced germination in response to ethanol treatment.
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Aconitum/metabolismo , Etanol/farmacología , Germinación/efectos de los fármacos , Proteínas de Plantas/análisis , Proteínas de Plantas/metabolismo , Semillas/efectos de los fármacos , Semillas/crecimiento & desarrollo , Aconitum/crecimiento & desarrollo , Especies en Peligro de Extinción , Plantas Medicinales/metabolismoRESUMEN
INTRODUCTION: Metabolomics is an 'omics' approach that aims to comprehensively analyse all metabolites in a biological sample, and has great potential for directly elucidating plant metabolic processes. Increasing evidence supports the view that plants produce a broad range of low-molecular-weight secondary metabolites responsible for variation from species to species, thus enabling the use of secondary metabolite profiling in the chemotaxonomy. OBJECTIVE: To gain deeper insights into the metabolites to increasing plant diversity, we performed systematic untargeted metabolite profiling to exploit the different parts and species of Aconitum as a case study. METHOD: Application of ultraperformance liquid chromatography-quadrupole time-of-flight-high-definition mass spectrometry (UPLC-QTOF-HDMS) equipped with electrospray ionisation and coupled with pattern recognition analyses to study constituents in the root of two kinds of Aconitum species. RESULTS: Twenty-two metabolites between the mother root of Aconitum carmichaelii Debx (CHW) and lateral root of Aconitum carmichaelii Debx (SFZ) and 13 metabolites between the CHW and root of Aconitum kusnezoffii Reichb (CW) have been identified. Of note, songorine, carmichaeline and isotalatizidine did not exist in CW, whereas they are present in the SFZ and CHW. CONCLUSION: Metabolomics based UPLC-QTOF-HDMS with multivariate statistical models was effective for analysis of constituents in the root of two kinds of Aconitum species.