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1.
Proc Natl Acad Sci U S A ; 120(52): e2311995120, 2023 Dec 26.
Artículo en Inglés | MEDLINE | ID: mdl-38113266

RESUMEN

Neurons in the brain communicate with each other at their synapses. It has long been understood that this communication occurs through biochemical processes. Here, we reveal that mechanical tension in neurons is essential for communication. Using in vitro rat hippocampal neurons, we find that 1) neurons become tout/tensed after forming synapses resulting in a contractile neural network, and 2) without this contractility, neurons fail to fire. To measure time evolution of network contractility in 3D (not 2D) extracellular matrix, we developed an ultrasensitive force sensor with 1 nN resolution. We employed Multi-Electrode Array and iGluSnFR, a glutamate sensor, to quantify neuronal firing at the network and at the single synapse scale, respectively. When neuron contractility is relaxed, both techniques show significantly reduced firing. Firing resumes when contractility is restored. This finding highlights the essential contribution of neural contractility in fundamental brain functions and has implications for our understanding of neural physiology.


Asunto(s)
Neuronas , Sinapsis , Ratas , Animales , Neuronas/fisiología , Sinapsis/fisiología , Hipocampo , Redes Neurales de la Computación , Encéfalo/fisiología , Potenciales de Acción/fisiología , Modelos Neurológicos
2.
Plant Cell Physiol ; 64(7): 771-785, 2023 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-37098222

RESUMEN

Artemisinin, a sesquiterpene lactone obtained from Artemisia annua, is an essential therapeutic against malaria. YABBY family transcription factor AaYABBY5 is an activator of AaCYP71AV1 (cytochrome P450-dependent hydroxylase) and AaDBR2 (double-bond reductase 2); however, the protein-protein interactions of AaYABBY5, as well as the mechanism of its regulation, have not yet been elucidated. AaWRKY9 protein is a positive regulator of artemisinin biosynthesis that activates AaGSW1 (glandular trichome-specific WRKY1) and AaDBR2 (double-bond reductase 2). In this study, YABBY-WRKY interactions are revealed to indirectly regulate artemisinin production. AaYABBY5 significantly increased the activity of the luciferase (LUC) gene fused to the promoter of AaGSW1. Toward the molecular basis of this regulation, AaYABBY5 interaction with AaWRKY9 protein was found. The combined effectors AaYABBY5 + AaWRKY9 showed synergistic effects toward the activities of AaGSW1 and AaDBR2 promoters, respectively. In AaYABBY5 overexpression plants, the expression of GSW1 was found to be significantly increased when compared to that of AaYABBY5 antisense or control plants. In addition, AaGSW1 was identified as an upstream activator of AaYABBY5. Further, it was found that AaJAZ8, a transcriptional repressor of jasmonate signaling, interacted with AaYABBY5 and attenuated its activity. Co-expression of AaYABBY5 and anti-AaJAZ8 in A. annua increased the activity of AaYABBY5 toward artemisinin biosynthesis. This current study provides the first indication of the molecular basis of regulation of artemisinin biosynthesis through YABBY-WRKY interactions, which are regulated through AaJAZ8. This knowledge presents AaYABBY5 overexpression plants as a powerful genetic resource for artemisinin biosynthesis.


Asunto(s)
Artemisia annua , Artemisininas , Artemisia annua/genética , Artemisia annua/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Regiones Promotoras Genéticas/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Artemisininas/metabolismo
3.
Crit Rev Biotechnol ; : 1-16, 2023 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-37380353

RESUMEN

Microalgae are the preferred species for producing astaxanthin because they pose a low toxicity risk than chemical synthesis. Astaxanthin has multiple health benefits and is being used in: medicines, nutraceuticals, cosmetics, and functional foods. Haematococcus pluvialis is a model microalga for astaxanthin biosynthesis; however, its natural astaxanthin content is low. Therefore, it is necessary to develop methods to improve the biosynthesis of astaxanthin to meet industrial demands, making its commercialization cost-effective. Several strategies related to cultivation conditions are employed to enhance the biosynthesis of astaxanthin in H. pluvialis. However, the mechanism of its regulation by transcription factors is unknown. For the first time, this study critically reviewed the studies on identifying transcription factors, progress in H. pluvialis genetic transformation, and use of phytohormones that increase the gene expression related to astaxanthin biosynthesis. In addition, we propose future approaches, including (i) Cloning and characterization of transcription factors, (ii) Transcriptional engineering through overexpression of positive regulators or downregulation/silencing of negative regulators, (iii) Gene editing for enrichment or deletion of transcription factors binding sites, (iv) Hormonal modulation of transcription factors. This review provides considerable knowledge about the molecular regulation of astaxanthin biosynthesis and the existing research gap. Besides, it provides the basis for transcription factors mediated metabolic engineering of astaxanthin biosynthesis in H. pluvialis.

4.
Molecules ; 26(22)2021 Nov 18.
Artículo en Inglés | MEDLINE | ID: mdl-34834066

RESUMEN

Angiotensin converting enzyme (ACE) plays a crucial role in regulating blood pressure in the human body. Identification of potential ACE inhibitors from medicinal plants supported the idea of repurposing these medicinal plants against hypertension. A method based on ultra-performance liquid chromatography (UPLC) coupled with a diode array detector (DAD) was used for the rapid screening of plant extracts and purified compounds to determine their ACE inhibitory activity. Hippuryl-histidiyl-leucine (HHL) was used as a substrate, which is converted into hippuric acid (HA) by the action of ACE. A calibration curve of the substrate HHL was developed with the linear regression 0.999. The limits of detection and quantification of this method were found to be 0.134 and 0.4061 mM, respectively. Different parameters of ACE inhibitory assay were optimized, including concentration, incubation time and temperature. The ACE inhibition potential of Adhatoda vasica (methanolic-aqueous extract) and its isolated pyrroquinazoline alkaloids, vasicinol (1), vasicine (2) and vasicinone (3) was evaluated. Compounds 1-3 were characterized by various spectroscopic techniques. The IC50 values of vasicinol (1), vasicine (2) and vasicinone (3) were found to be 6.45, 2.60 and 13.49 mM, respectively. Molecular docking studies of compounds 1-3 were also performed. Among these compounds, vasicinol (1) binds as effectively as captopril, a standard drug of ACE inhibition.


Asunto(s)
Alcaloides/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Género Justicia/química , Extractos Vegetales/farmacología , Quinazolinas/farmacología , Alcaloides/química , Inhibidores de la Enzima Convertidora de Angiotensina/química , Cromatografía Líquida de Alta Presión , Descubrimiento de Drogas , Evaluación Preclínica de Medicamentos , Humanos , Simulación del Acoplamiento Molecular , Extractos Vegetales/química , Quinazolinas/química
5.
Ecotoxicol Environ Saf ; 206: 111202, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-32889311

RESUMEN

Aim of the current study was to investigate the effect of exogenously inoculated root endophytic fungus, Piriformospora indica, on molecular, biochemical, morphological and physiological parameters of Artemisia annua L. treated with different concentrations (0, 50, 100 and 150 µmol/L) of arsenic (As) stress. As was significantly accumulated in the roots than shoots of P. indica-inoculated plants. As accumulation and immobilization in the roots is directly associated with the successful fungal colonization that restricts most of As as compared to the aerial parts. A total of 4.1, 11.2 and 25.6 mg/kg dry weight of As was accumulated in the roots of inoculated plants supplemented with 50, 100 and 150 µmol/L of As, respectively as shown by atomic absorption spectroscopy. P. indica showed significant tolerance in vitro to As toxicity even at high concentration. Furthermore, flavonoids, artemisinin and overall biomass were significantly increased in inoculated-stressed plants. Superoxide dismutase and peroxidase activities were increased 1.6 and 1.2 fold, respectively under 150 µmol/L stress in P. indica-colonized plants. Similar trend was followed by ascorbate peroxidase, catalase and glutathione reductase. Like that, phenolic acid and phenolic compounds showed a significant increase in colonized plants as compared to their respective control/un-colonize stressed plants. The real-time PCR revealed that transcriptional levels of artemisinin biosynthesis genes, isoprenoids, terpenes, flavonoids biosynthetic pathway genes and signal molecules were prominently enhanced in inoculated stressed plants than un-inoculated stressed plants.


Asunto(s)
Arseniatos/metabolismo , Artemisia annua/metabolismo , Basidiomycota/metabolismo , Raíces de Plantas/metabolismo , Antioxidantes/metabolismo , Arseniatos/toxicidad , Artemisia annua/efectos de los fármacos , Artemisia annua/genética , Artemisia annua/microbiología , Artemisininas/metabolismo , Ascorbato Peroxidasas/metabolismo , Basidiomycota/crecimiento & desarrollo , Biomasa , Relación Dosis-Respuesta a Droga , Modelos Teóricos , Presión Osmótica/efectos de los fármacos , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Raíces de Plantas/microbiología , Transcripción Genética/efectos de los fármacos
6.
Metabolomics ; 15(9): 116, 2019 08 22.
Artículo en Inglés | MEDLINE | ID: mdl-31440842

RESUMEN

INTRODUCTION: Polyherbal formulations are an integral part of various indigenous medicinal systems such as Traditional Chinese Medicine (TCM) and Ayurveda. The presence of a very large number of compounds makes the quality control of polyherbal formulations very difficult. OBJECTIVES: To overcome this problem, we have developed a comprehensive strategy for the dereplication of natural products in polyherbal formulations by using Adhatoda vasica as a case study. METHODS: The strategy is based on five major steps: the collection of plant samples from different locations to observe the effects of environmental variables; LC-ESI-MS/MS-based untargeted metabolite profiling of the plant samples to identify marker compounds using extensive chemometric analysis of the obtained data; the identification of marker compounds in polyherbal products; the isolation, purification and characterization of the marker compounds; and MRM-based quantitative analysis of the isolated marker compounds using LC-ESI-MS/MS. RESULTS: Using this strategy, we identified a total of 51 compounds in the methanolic extract of A. vasica plants from 14 accessions. Chemical fingerprinting of the plant led to the identification of characteristic peaks that were used to confirm the presence of A. vasica in complex polyherbal formulations. Four quinazoline alkaloids (marker compounds) were isolated, purified and quantified in various herbal formulations containing A. vasica. CONCLUSION: This method demonstrates a comprehensive strategy based on untargeted and targeted metabolite analysis that can be used for the standardization of complex polyherbal formulations.


Asunto(s)
Género Justicia/química , Metabolómica/métodos , Extractos Vegetales/normas , Espectrometría de Masa por Ionización de Electrospray/métodos , Metabolómica/normas , Extractos Vegetales/química , Plantas Medicinales/química , Espectrometría de Masa por Ionización de Electrospray/normas
7.
Acta Biochim Biophys Sin (Shanghai) ; 51(3): 229-242, 2019 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-30883651

RESUMEN

The beneficial endophytic microorganisms have received significant attention in agriculture because of their exceptional capabilities to facilitate functions like nutrient enrichment, water status, and stress tolerance (biotic and abiotic). This review signifies the molecular mechanisms to better understand the Piriformospora indica-mediated plants improvement or protection for sustainable agriculture. P. indica, an endophytic fungus, belonging to the order Sebacinales (Basidiomycota), is versatile in building mutualistic associations with a variety of plants including pteridophytes, bryophytes, gymnosperms, and angiosperms. P. indica has enormous potential to manipulate the hormonal pathway such as the production of indole-3-acetic acid which in turn increases root proliferation and subsequently improves plant nutrient acquisition. P. indica also enhances components of the antioxidant system and expression of stress-related genes which induce plant stress tolerance under adverse environmental conditions. P. indica has tremendous potential for crop improvement because of its multi-dimensional functions such as plant growth promotion, immunomodulatory effect, biofertilizer, obviates biotic (pathogens) and abiotic (metal toxicity, water stress, soil structure, salt, and pH) stresses, phytoremediator, and bio-herbicide. Considering the above points, herein, we reviewed the physiological and molecular mechanisms underlying P. indica-mediated plants improvement or protection under diverse agricultural environment. The first part of the review focuses on the symbiotic association of P. indica with special reference to biotic and abiotic stress tolerance and host plant root colonization mechanisms, respectively. Emphasis is given to the expression level of essential genes involved in the processes that induce changes at the cellular level. The last half emphasizes critical aspects related to the seed germination, plant yield, and nutrients acquisition.


Asunto(s)
Agricultura , Basidiomycota/fisiología , Endófitos/fisiología , Desarrollo de la Planta , Crecimiento Sostenible , Germinación , Raíces de Plantas/microbiología , Estrés Fisiológico
8.
Adv Sci (Weinh) ; 11(11): e2306826, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38161217

RESUMEN

Motivated by the unexplored potential of in vitro neural systems for computing and by the corresponding need of versatile, scalable interfaces for multimodal interaction, an accurate, modular, fully customizable, and portable recording/stimulation solution that can be easily fabricated, robustly operated, and broadly disseminated is presented. This approach entails a reconfigurable platform that works across multiple industry standards and that enables a complete signal chain, from neural substrates sampled through micro-electrode arrays (MEAs) to data acquisition, downstream analysis, and cloud storage. Built-in modularity supports the seamless integration of electrical/optical stimulation and fluidic interfaces. Custom MEA fabrication leverages maskless photolithography, favoring the rapid prototyping of a variety of configurations, spatial topologies, and constitutive materials. Through a dedicated analysis and management software suite, the utility and robustness of this system are demonstrated across neural cultures and applications, including embryonic stem cell-derived and primary neurons, organotypic brain slices, 3D engineered tissue mimics, concurrent calcium imaging, and long-term recording. Overall, this technology, termed "mind in vitro" to underscore the computing inspiration, provides an end-to-end solution that can be widely deployed due to its affordable (>10× cost reduction) and open-source nature, catering to the expanding needs of both conventional and unconventional electrophysiology.


Asunto(s)
Encéfalo , Neuronas , Electrodos , Encéfalo/fisiología , Neuronas/fisiología , Estimulación Eléctrica , Fenómenos Electrofisiológicos/fisiología
9.
Front Pharmacol ; 13: 797853, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35422700

RESUMEN

Fetal hemoglobin (HbF) is a potent genetic modifier, and the γ-globin gene induction has proven to be a sustainable therapeutic approach for the management of ß-thalassemia. In this study, we have evaluated the HbF induction ability of A. vasica in vitro and in vivo, and the identification of potential therapeutic compounds through a bioassay-guided approach. In vitro benzidine-Hb assay demonstrated strong erythroid differentiation of K562 cells by A. vasica extracts. Subsequently, an in vivo study with an aqueous extract of A. vasica (100 mg/kg) showed significant induction of the γ-globin gene and HbF production. While in the acute study, the hematological and biochemical indices were found to be unaltered at the lower dose of A. vasica. Following the bioassay-guided approach, two isolated compounds, vasicinol (1) and vasicine (2) strongly enhanced HbF levels and showed prominent cellular growth kinetics with ample accumulation of total hemoglobin in K562 cultures. High HbF levels were examined by immunofluorescence and flow cytometry analysis, concomitant with the overexpression in the γ-globin gene level. Compound 1 (0.1 µM) and compound 2 (1 µM) resulted in a greater increase in F-cells (90 and 83%) with marked up (8-fold and 5.1-fold) expression of the γ-globin gene, respectively. Molecular docking studies indicated strong binding affinities of (1) and (2) with HDAC2 and KDM1 protein that predict the possible mechanism of compounds in inhibition of these epigenetic regulators in the γ-globin gene reactivation. Altogether, these observations demonstrated the therapeutic usefulness of A. vasica for fostering HbF production in clinical implications for blood disorders.

10.
Hortic Res ; 8(1): 257, 2021 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-34848710

RESUMEN

Artemisia annua is a medicinal plant rich in terpenes and flavonoids with useful biological activities such as antioxidant, anticancer, and antimalarial activities. The transcriptional regulation of flavonoid biosynthesis in A. annua has not been well-studied. In this study, we identified a YABBY family transcription factor, AaYABBY5, as a positive regulator of anthocyanin and total flavonoid contents in A. annua. AaYABBY5 was selected based on its similar expression pattern to the phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), and flavonol synthase (FLS) genes. A transient dual-luciferase assay in Nicotiana bethamiana with the AaYABBY5 effector showed a significant increase in the activity of the downstream LUC gene, with reporters AaPAL, AaCHS, AaCHI, and AaUFGT. The yeast one-hybrid system further confirmed the direct activation of these promoters by AaYABBY5. Gene expression analysis of stably transformed AaYABBY5 overexpression, AaYABBY5 antisense, and control plants revealed a significant increase in the expression of AaPAL, AaCHS, AaCHI, AaFLS, AaFSII, AaLDOX, and AaUFGT in AaYABBY5 overexpression plants. Moreover, their total flavonoid content and anthocyanin content were also found to increase. AaYABBY5 antisense plants showed a significant decrease in the expression of flavonoid biosynthetic genes, as well as a decrease in anthocyanin and total flavonoid contents. In addition, phenotypic analysis revealed deep purple-pigmented stems, an increase in the leaf lamina size, and higher trichome densities in AaYABBY5 overexpression plants. Together, these data proved that AaYABBY5 is a positive regulator of flavonoid biosynthesis in A. annua. Our study provides candidate transcription factors for the improvement of flavonoid concentrations in A. annua and can be further extended to elucidate its mechanism of regulating trichome development.

11.
Metabolites ; 11(8)2021 Jul 28.
Artículo en Inglés | MEDLINE | ID: mdl-34436430

RESUMEN

The discovery and identification of novel natural products of medicinal importance in the herbal medicine industry becomes a challenge. The complexity of this process can be reduced by dereplication strategies. The current study includes a method based on high-performance liquid chromatography (HPLC), using the evaporative light scattering detector (ELSD) to identify the 12 most common secondary metabolites in plant extracts. Twelve compounds including rutin, taxifolin, quercetin, apigenin, kaempferol, betulinic acid, oleanolic acid, betulin, lupeol, stigmasterol, and ß-sitosterol were analyzed simultaneously. The polarity of the compounds varied greatly from highly polar (flavonoids) to non-polar (triterpenes and sterols). This method was also tested for HPLC-DAD and HPLC-ESI-MS/MS analysis. Oleanolic acid and ursolic acid could not be separated in HPLC-ELSD analysis but were differentiated using LC-ESI-MS/MS analysis due to different fragment ions. The regression values (R2 > 0.996) showed good linearity in the range of 50-1000 µg/mL for all compounds. The range of LOD and LOQ values were 7.76-38.30 µg/mL and 23.52-116.06 µg/mL, respectively. %RSD and % trueness values of inter and intraday studies were mostly <10%. This method was applied on 10 species of medicinal plants. The dereplication strategy has the potential to facilitate and shorten the identification process of common secondary metabolites in complex plant extracts.

12.
J Pharm Biomed Anal ; 178: 112918, 2020 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-31629227

RESUMEN

Ziziphus jujuba and Ziziphus nummularia are two important species of the genus Ziziphus. Both plants offer great commercial value and are consumed as food around the world and used for their medicinal values such as anti-inflammatory, antioxidant and hepatoprotective activities. Comprehensive chemical profiling of Z. jujuba and Z. nummularia was done through identification of major metabolites using HPLC-QTOF-MS/MS and quantification of eight analytes using HPLC-IonTrap-MS/MS analysis. A total of 53 compounds were identified from their high-resolution mass spectra in both positive and negative ionization modes. Among these, 52 compounds were found to be present in Z. jujuba, and 45 in Z. nummularia. Chemometric analysis was also performed to assess the distribution of identified compounds and to determine how the obtained data can be used to discriminate between the two species. Moreover, a method for the quantification of eight analytes including, 6″'-feruloylspinosin (1), apigenin (2), apigenin-7-O-glucoside (3), catechin (4), jujuboside A (5), jujuboside B (6), luteolin (7) and quercetin (8) was developed. The method expressed excellent accuracy with less than 3% error and good reproducibility with less than 4% RSD. The limit-of-detection (LOD) was also found to be very low ranging between 0.06 ng/mL to 4.10 ng/mL while limit-of-quantitation (LOQ) values were in the range of 0.17 ng/mL to 12.42 ng/mL. The analyte concentrations were found to be varying from 1.32 mg/kg to 645.76 mg/kg in both species. The developed method was used to identify and quantify marker compounds in fruit and whole plant samples of both species and in their herbal products as well. The present work is unprecedented, as there is no such extensive study targeting fruits, leaves and herbal formulations together using high-resolution techniques. The study will provide great utility in drug discovery, in taxonomical identification of these plants and to develop quality control protocols for their herbal formulations.


Asunto(s)
Ziziphus/química , Ziziphus/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Correlación de Datos , Frutas/química , Frutas/metabolismo , Extractos Vegetales/química , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Quercetina/química , Reproducibilidad de los Resultados , Saponinas/química , Espectrometría de Masas en Tándem/métodos
13.
J Ayub Med Coll Abbottabad ; 21(3): 72-5, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-20929018

RESUMEN

BACKGROUND: Old World Cutaneous Leishmaniasis (OWCL) is a preventable skin infection that leads to morbidity and social isolation. It is spreading rapidly. The sore of OWCL may be a non-ulcerative red papule, nodule or a large mutilating ulcer. The ulcer is typically painless and can leave a disfiguring scar. METHODS: This was a descriptive study. The diagnosis of OWCL was established by finding LD bodies in skin smear preparation. RESULTS: This study identified 1680 cutaneous leishmaniasis in 1767 skin ulcers. Children (n = 924) were infected more than other age groups (n = 756). There were typical skin sore of OWCL in 1512 cases while 168 patients had atypical presentation. The ulcers were painless in 1603 patients. History of insect bite was present in 1366 cases, thorn prick in 156 patients, religious visit to endemic areas in 256 patients, and 4 patients had post surgical non healing wound. Lesions with 4 to 6 months of age had a maximum yield of LD bodies. There were 498 patients from different areas of Peshawar; 688 cases from leishmania endemic belt of FATA while 89 patients came from other urban and rural areas of NWFP. CONCLUSIONS: There is a tremendous increase in cases of OWCL and the disease became endemic in many regions of Pakistan. The bordering areas along Afghanistan have constituted an endemic belt that had invaded the neighboring urban and rural areas. Several chronic non healing ulcers had been diagnosed as OWCL. Many cases have been detected in Peshawar. People need education about the nature of the diseases and the efficacy of personal protective measures. Spray with suitable insecticides is required in all residential areas.


Asunto(s)
Leishmaniasis Cutánea/epidemiología , Úlcera Cutánea/parasitología , Adolescente , Adulto , Niño , Diagnóstico Diferencial , Femenino , Humanos , Leishmaniasis Cutánea/diagnóstico , Masculino , Persona de Mediana Edad , Pakistán/epidemiología , Úlcera Cutánea/diagnóstico , Úlcera Cutánea/epidemiología
14.
Front Plant Sci ; 10: 1084, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31552076

RESUMEN

Artemisinin is an effective antimalarial sesquiterpene lactone synthesized in Artemisia annua. Various transcription factors have been previously reported that can influence the biosynthesis of artemisinin; however, the effect of YABBY family transcription factors on artemisinin biosynthesis was unknown. In the present study, we cloned and characterized AaYABBY5: a homolog of MsYABBY5 in Mentha spicata which is involved in modulating the monoterpenes, as a positive regulator of artemisinin biosynthesis in A. annua. AaYABBY5 was found localized to the nucleus, and its expression was found to be induced by exogenous methyl jasmonic acid (MeJA) treatment. In the dual-luciferase reporter assay, it was found that AaYABBY5 significantly increased the activities of promoters of amorpha-4,11-diene synthase (ADS), cytochrome P450 monooxygenase (CYP71AV1), double-bond reductase 2 (DBR2), and aldehyde dehydrogenase 1 (ALDH1) genes. Yeast one hybrid assay showed that AaYABBY5 directly bonds to the promoters of CYP71AV1 and DBR2 genes. Quantitative real-time polymerase chain reaction (qPCR) of AaYABBY5 overexpression and AaYABBY5 antisense plants revealed a significant increase in the expression of ADS, CYP71AV1, DBR2, and ALDH1 in AaYABBY5 overexpression plants and a significant decrease in the expression of these genes in AaYABBY5 antisense A. annua, respectively. Furthermore, the results of high-performance liquid chromatography (HPLC) showed that the artemisinin and its precursor dihydroartemisinic acid were significantly increased in the AaYABBY5 overexpression plants while AaYABBY5 downregulation resulted in a significant decrease in the concentration of artemisinin. Taken together, these results explicitly represent that AaYABBY5 is a positive regulator of artemisinin biosynthesis in A. annua.

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