Resistance of Microbial Community of Artemisia annua L.to Pathogenic Fungi

[Objectives]This paper was to figure out whether the dominant bacterial community has the role and effect of bacterial community and its defense mechanism against potential pathogenic fungi in Artemisia annua,and thus establish a systematic model of bacteria-fungus-plant.[Methods]Fifty-eight strains of bacteria and one strain of pathogenic fungi,Globisporangium ultimatum,were used for the experiments.These 58 bacterial strains were assembled into a bacterial community,and the bacteria with abundance in the top 1%were reassembled into a dominant bacterial community as measured by 16S rDNA.[Results]The growth of A.annua seedlings inoculated with bacterial communities and pathogenic fungi or dominant bacterial communities and pathogenic fungi was significantly better than that of A.annua seedlings inoculated with pathogenic fungi during in vitro confrontation,which was evident in both enzymatic and non-enzymatic antioxidant assays.[Conclusions]The results suggest that the dominant bacterial community has a crucial role as a representative core microbial community of synthetic bacterial community,which can protect plants by interfering with the growth of phytopathogenic fungi mediated by chemical signals,and can be used as the main synthetic community of biocides to achieve the effect of biocontrol.

Effect of Artemisia annua (Asteraceae) Extracts on Hemolysis in Individuals with G6PD-Deficiency

Individuals with Glucose-6-phosphate dehydrogenase (G6PD) deficiency are susceptible to hemolytic anemia when exposed to pro-oxidant substances. This study investigates the hemolytic impact of Artemisia annua (A. annua) extracts in G6PD-deficient subjects through a mixed experimental approach. In the in vitro phase, red blood cells from G6PD-deficient individuals and rats induced with Dehydroepiandrosterone (DHEA) were exposed to various concentrations of A. annua infusion, with distilled water and physiological saline as positive and negative controls respectively. The in vivo study involved G6PD-deficient Wistar rats divided into three groups receiving A. annua infusion, quinine (positive control), and distilled water (negative control) via gavage. Blood samples were collected for biochemical and hematological analyses. Notably, at a 40% concentration of A. annua infusion, there was a significant increase in the hemolysis rate of G6PD-deficient red blood cells compared to controls (p A. annua exhibited elevated aspartate aminotransferase (129.25 ± 4.55 U/L vs. 80.09 ± 4.03 U/L;p A. annua infusion tested positive for saponins. These findings underscore the risk of hemolysis in G6PD-deficient individuals upon ingesting A. annua.

Chemotaxonomic Study of the Covid-Organics of Madagascar Based on the Chemical Composition of Their Essential Oils

The aim of this study was to investigate the chemical composition of the essential oils of “Covid-Organics” of Madagascar (62% Artemisia annua and two other undisclosed medicinal plants) used as curative and preventive treatments of Covid-19, to identify its constituent species. The essential oils isolated by hydro-distillation from two samples (curative and preventive) were analyzed by GC-FID and GC-MS. These essential oils (curative and preventive) were mainly dominated by camphor (17.9% and 11.9%, respectively), spathulenol (4.8% and 11.8%, respectively), α-acorenol (4.3% and 3.7%, respectively), (E)-β-caryophyllene (3.4% and 4.2%, respectively), 1,8-cineole (3.1% and 3.6%, respectively), hexadecanoïc acid (3.8% and 3.2%, respectively) and caryophyllene oxide (3.4% and 2.4%, respectively). From the chemical composition, two species were identified, A. annua characterised by camphor and Cinnamomumcamphora (Ravintsara) characterised by 1,8-cineole and sabinene. However, we were unable to identify the third species.

Effect of Organic Amendment on the Growth of Artemisia annua in the North of Côte d’Ivoire

Malaria causes many deaths around the world, particularly in Africa, which ultimately affects the socio-economic development of African countries. The resistance of Plasmodium falciparum to quinine-based drugs led to new studies showing the efficiency of new artemisin-based drugs. The molecule artemisin is extracted from Artemisia annua a plant from China that has been used for decades in traditional Chinese medicine. The purpose of this study is to improve the production of sweet wormwood (Artemisia annua) using organic fertilizers in the north of Cote d’Ivoire. To do so, a morpho-pedological characterization of the study site was firstly performed to determine the soil type and their fertility level. Then, a randomized complete block system including two factors (the quantity of compost and the plant density) was implemented to test the effect of organic amendment and plant arrangement on the growth of Artemisia annua. Six treatments were set up: a control plot (no compost) where the plants are arranged in square (T0D1) and the plants are arranged in staggered (T0D2). Then, a treatment with compost addition of 25 t/ha where the plants are arranged in square (T1D1) and in staggered (T1D2). A treatment with compost addition of 50 t/ha where plants are arranged in square (T2D1) and in staggered (T2D2). Our results showed that the soils hosting our experimentation are Arenithic Plinthic Ferrasols with a very low level of fertility, prone to leaching and erosion. T1D2 and T2D2 treatments obtained the highest yields of 2.82 t/ha and 3.91 t/ha, respectively. Our findings indicate that a high dose of organic amendment combined with a staggered plant arrangement strongly improves the biomass production of sweet wormwood. This is in agreement with previous studies showing that the addition of organic matter can restore the level of soil fertility by increasing soil porosity and the activity of micro and macroorganisms.

Effects of Fungal Elicitors on Cell Growth and Artemisinin Accumulation in Hairy Root Cultures of Artemisia annua

The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr.) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae , the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.

Elicitation on Artemisinin Biosynthesis in Artemisia annua Hairy Roots by the Oligosaccharide Extract from the Endophytic Colletotrichum sp. B501

The oligosaccharide elicitor from the mycelial wall of an endophytic Colletotrichum sp. B501 promoted the production of artemisinin in Artemisia annua L. hairy root culture. When hairy roots of 22-day-old cultures (later growth phase) were exposed to the elicitor (20 mg/L) for 4 d, the maximum content of artemisinin reached 1.15 mg/g, a 64.29% increment over the control. The electron X-ray microanalysis disclosed the rapid accumulation of Ca 2+ in the elicited cortical cells of hairy root. The electronic microscope observation revealed the high electron density area in vacuole of elicited cells. During the first day of elicitation the peroxidase activity of hairy roots was improved sharply. Some cellular morphological changes including cell shrinkage, condensation of cytoplasm and nuclear fragmentation, coincident with the appearance of DNA ladders, were observed after the third day of elicitation. It was suggested that the oligosaccharide elicitor triggered the programmed cell death, which may provide the substance or chemical signal for artemisinin biosynthesis.

Molecular Cloning, Escherichia coli Expression and Genomic Organization of Squalene Synthase Gene from Artemisia annua

A 1 539 by squalene synthase (AaSQS) cDNA was cloned from a high-yield Artemisia annua L. strain 001 by reverse transcription-polymerise chain reaction (RT-PCR). The amino acid sequence of AaSQS is 70%, 77%, 44% and 39%a identical to that of squalene synthases from Arabidopsis thaliana, tobacco, human and yeast, respectively. The AaSQS genomic DNA has a complex organization containing 14 exons and 13 introns. Full-length or C-terminal truncated cDNA was subcloned into prokaryotic expression vector pET30a and the constructed plasmid was introduced to Escherichia coli strain BL21 (DE3) for induced overexpression. No squalene synthase protein with expected molecular mass was observed in E. cola containing the putative full-length squalene synthase cDNA, however, overexpression in E. coli was achieved by truncating 30 amino acids of hydrophobic region at the carboxy terminus.

Cloning, E. coli Expression and Molecular Analysis of a Novel Sesquiterpene Synthase Gene from Artemisia annua

A 1 886 bp full-length sesquiterpene synthase (AaSES) cDNA was cloned front a high-yield Artemisia annua L. strain 001 by a rapid amplification of cDNA end (RACE) strategy. AaSES is 59% identical to Artemisia cyclase cDNA clone cASC125, 50% identical to epi-cedrol synthase from A. annua , 48% identical to amorpha-4, 11-diene synthase from A. annua, 39% identical to the 5-epi-aristolechene synthase from tabacco, 38 % identical to vetispiradiene synthase front H. muticus, 41 % identical to the, delta-cadinene synthase from cotton. The coding region of the cDNA was inserted into a procaryotic expression vector pET-30a and overexpressed in E. coli BL21 ( DE3). The cyclase proteins extracted front bacterial culture were found largely in an insoluble protein fraction. AaSES expresses in leaves, stems a-rid flowers, not in roots as indicated by Northern blotting analysis.

Can Yin-Chai-Xiao-Du decoction be useful of COVID-19?the mechanism research based on network pharmacology

Background:In this study,we preliminarily investigated the mechanism of Yin-Chai-Xiao-Du decoction for the treatment of COVID-19 by the method of network pharmacology.Methods:The potential targets and pathways of Yin-Chai-Xiao-Du decoction for the treatment of COVID-19 were examined using network pharmacology;the ingredient and active targets of Yin-Chai-Xiao-Du decoction were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and PharmMapper databases;the COVID-19-related targets were obtained from the online Mendelian inheritance in man,GeneCards,and GeneMANIA databases;the STRING database and Cytoscape were used to build a protein-protein interaction network,and a Network Analyzer tool was used to perform topology analysis to screen for the key ingredients and targets;the ClueGO and KOBAS 3.0 databases were for the enrichment analysis of gene function(Gene Oncology)and gene pathway(Kyoto Encyclopedia of Genes and Genomes);the herb-ingredient-target-pathway network diagram was constructed by Cytoscape.Results:The core herbs screened by the network pharmacological analysis were Jinyinhua(Lonicerae japonicae flos),Lianqiao(Forsythia suspensa),Chaihu(Bupleuri radix),Huangqin(Scutellariae radix),Yinchen(Herba Artemisiae Scopariae),Guanghuoxiang(Pogostemonis herba),Roudoukou(Semen myristicae)and Qinghao(Artemisiae annuae herba).A total of 293 active ingredients were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the key ingredients were quercetin,kaempferol,isorhamnetin,stigmasterol,beta-sitosterol,and luteolin.Yin-Chai-Xiao-Du decoction has 138 COVID-19-related targets,and the key targets were mitogen-activated protein kinase 3,interleukin-6,tumor necrosis factor,vascular endothelial growth factor A,and CC motif ligand 2.Kyoto Encyclopedia of Genes and Genomes analysis revealed 120 enriched gene pathways,and the key pathways were signaling by interleukins,immune system,cytokine signaling in the immune system,and the signaling pathways of interleukin-17,tumor necrosis factor,and relaxin.Conclusion:The core herbs of Yin-Chai-Xiao-Du decoction are Jinyinhua(Lonicerae japonicae flos),Lianqiao(Forsythia suspensa),Chaihu(Bupleuri radix),Huangqin(Scutellariae radix),Yinchen(Herba Artemisiae Scopariae),Guanghuoxiang(Pogostemonis herba),Roudoukou(Semen myristicae)and Qinghao(Artemisiae annuae herba).The key ingredients are quercetin,kaempferol,isorhamnetin,stigmasterol,and beta-sitosterol;the critical targets are luteolin,interleukin-6,mitogen-activated protein kinase 3,tumor necrosis factor,and CC motif ligand 2;and the core signaling pathways are those mediated by interleukin-17,tumor necrosis factor,and relaxin.

Effects of Ag-carrying Zirconium Phosphate on the Kinetics of Growth of the Roots of Culture Artemisia annua

在植物组织和细胞培养过程中 ,尤其是在生物反应器培养中的染菌问题 ,一直是制约植物细胞培养工业化的难题。通过比较各种防腐剂的抑菌效果 ,确定银型磷酸锆盐抗菌粉为青蒿根培养的最佳防腐剂。银型磷酸锆盐抗菌粉在浓度为 30mg/L时 ,既能降低培养液的染菌几率 ,又不明显抑制青蒿根的生长及青蒿素的生物合成。在添加 30mg/L抗菌粉的培养液中进行的青蒿根生长、pH值变化以及残糖、铵离子和硝酸根离子消耗的动力学研究表明 ,在 4 0d内青蒿根在培养液中生长良好 ,营养成分的消耗和对照呈相似的趋势。

Studies on Acaricidal Bioactivities of Artemisia annua L.Extracts Against Tetranychus cinnabarinus Bois.(Acari:Tetranychidae)

The aim of this study was to determine the best extraction technique, the most suitable solvent, the optimal plant parts, and the acaricidal activities of Artemisia annua L. The petroleum ether （30-60℃）, petroleum ether （60-90℃）, ethanol, acetone, and water parallel and sequenced extracts were obtained from the leaves, stems and roots of different period of A. annua L. in April, May, June, July and September respectively. And then the acaricidal bioactivities against Tetranychus cinnabarinus of all extracts were determined by the slide-capillary method in the laboratory. The results indicated that the acaricidal bioactivities elevated as the development of A. annua plant at the concentration of 5 mg mL-L The general tendency exhibited the sequence of July 〉 June 〉 May 〉 April, but September decreased comparing to July. However, the most effective extracts in five months were all acetone parallel extract of A. annua leaf, and the corrected mortalities treated after 48 h ranged from 74 to 100%. The median lethal concentrations （LC50） against T. cinnabarinus of acetone parallel extracts ofA. annua leaves in September, July, June, May and April were 0.5986, 0.4341, 0.8376, 0.9443 and 1.3817 mg mL^-1, respectively, treated after 48 h. The 13 groups were isolated from acetone extracts ofA. annua leaves in July by column chromatography, both the 1 lth and 12th groups exhibited strong bioactivities. The median lethal concentrations of the 1 lth and 12th groups against T. cinnabarinus were 0.3683 and 0.1586 mg mL^-1, respectively. The acetone parallel extract ofA. annua leaf in July was the most toxic to T. cinnabarinus and the corrected mortality was 100% after 48 h. The acetone parallel extract of the 1 lth and 12th groupswere the most active components, acted as the emphases in further study.

Dried-leaf Artemisia annua: A practical malaria therapeutic for developing countries?

Artemisinin from the plant Artemisia annua （A. annua） L., and used as artemisinin combination therapy （ACT）, is the current best therapeutic for treating malaria, a disease that hits children and adults especially in developing countries. Traditionally, A. annua was used by the Chinese as a tea to treat “fever”. More recently, investiga-tors have shown that tea infusions and oral consumption of the dried leaves of the plant have prophylactic and therapeutic effcacy. The presence of a complex matrix of chemicals within the leaves seems to enhance both the bioavailability and effcacy of artemisinin. Although about 1000-fold less potent than artemisinin in their antiplasmodial activity, these plant chemicals are mainly small molecules that include other artemisinic compounds, terpenes （mainly mono and sesqui）, favonoids, and polyphenolic acids. In addition, polysaccharide constituents of A. an-nua may enhance bioavailability of artemisinin. Rodent pharmacokinetics showed longer T？ and Tmax and greater Cmax and AUC in Plasmodium chabaudi -infected mice treated with A. annua dried leaves than in healthy mice. Pharmacokinetics of deoxyartemisinin, a liver metabolite of artemisinin, was more inhibited in infected than in healthy mice. In healthy mice, artemisinin serum levels were 〉 40-fold greater in dried leaf fed mice than those fed with pure artemisinin. Human trial data showed that when delivered as dried leaves, 40-fold less artemisinin was required to obtain a therapeutic response compared to pure artemisinin. ACTs are still unaffordable for many malaria patients, and cost estimates for A. annua dried leaf tablet production are orders of magnitude less than for ACT, despite improvements in the production capacity. Considering that for 〉 2000 years this plant was used in traditional Chinese medicine for treatment of fever with no apparent appearance of artemisinin drug resistance, the evidence argues for inclusion of affordable A. annua dried leaf tablets into the arsenal of drugs to combat malaria and other artemisinin-susceptible diseases.

Maintaining the artemisinin content through direct and indirect in vitro regeneration and their assessment of variations with the field grown mother plants of Artemisia annua L.

An efficient protocol for maintaining the artemisinin content in tissue culture and high frequency of in vitro direct and indirect regenerations of multiple shoots of high artemisinin yielding genotypes of Artemisia annua has been developed and their comparison with field grown mother plant has been carried out.The eleven elite genotypes(containing more than 1%artemisinin)were tested on seven different modified media formulations.Modified half MS(Murashige and Skoog’s)media containing 100 mg L^(-1) myo-inositol,0.5 g L^(-1) casine hydrolysate,5 mg L^(-1) biotin,2 mL L^(-1) RT(Revised Tobacco)vitamin stock,0.5 mg L^(-1) BAP and 0.01 mg L^(-1) NAA showed best regeneration while,above modified MS medium containing 0.2 mg L^(-1) BAP and 0.2 mg L^(-1) NAA showed maximum shoot multiplication with maintained artemisinin content.Based on the chemical profiles of both the systems,minor difference was observed in their artemisinin content.A large scale culture of these plants maintained the normal growth index along with the artemisinin content and could be a better alternative to maintain the high artemisinin yielding genotypes with their genetic constraint in specific media combinations and also used as base material for further genetic improvement.

A new sesquiterpene from hairy root culture of Artemisia annua

A new sesquiterpene（Z）-7-acetoxy-methyl-11-methyl-3-methylenedodeca-1,6,10-triene（AMDT） was isolnted and identified from the methanol extract of the hairy root culture of Artemisia annua.The structure of AMDT was determined based on the analysis of spectroscopic data,notably of the 2D NMR spectra.This new compound showed cytotoxicity against human tumor cell lines 95-D and HeLa with IC50 values of 27.08 and 20.12μmol/L,respectively.

Growth Response of Artemisia annua by Effect of Types and Composition of Organic Fertilizer in Lowland

Artemisia annua is a plant used to cure malaria diseases. Artemisia plant contains artemisinin as secondary metabolite that used to eliminate parasite that caused malaria, such as Plasmodium falciparum. Artemisia growth affects production of artemisinin content in plant. Therefore, necessary environment conditions and appropriate organic manure application are needed to support the growth of Artemisia. This research aimed to determine the effect of fertilizer type and proportion in the medium on the Artemisia growth. This research was conducted at greenhouse of Faculty of Agriculture, Universitas Sebelas Maret, Surakarta, in October 2015 to January 2016. This research used a completely randomized design （CRD）, consisting of two factors of treatment with three replications. The first factor was type of fertilizer that consists of three types： horse manure fertilizer, compost filter press mud and cow manure fertilizer. The second factor was proportion of fertilizer with media consisted of five levels： fertilizer as media, proportion of fertilizer with media 4：1, 3：2, 2：3 and 1：4. Data were analyzed using analysis of variance and Duncan＇s multiple range test with level of 5%. It can be concluded that treatment with compost filter press mud provided the highest of plant height, root length, days to flowering, root volume, fresh weight and dry weight of crop.

Effect of Artemisia annua L. as Substitute to Sulfonamides (Sodium Sulfadimerzine) on Coccidiosis and Growth Performance in Rabbits

Coccidiosis is a disease caused by protozoa of the genus Eimeria which seriously affects young rabbits. Treatment based on the use of anticoccidial drugs is increasingly ineffective due to the rapid emergence of resistant strains of coccidia and the high cost of drugs. Consumer demand for rabbit products without chemical residues led to a growing interest in the use of medicinal plants as an alternative treatment for coccidiosis. The present study was carried out during the period of August to December 2020 to assess the anticoccidial effect of hydro-ethanolic extract of leaves of Artemisia annua L., in young rabbits. The antiparasitic efficacy of Artemisia extract was tested on 15 young rabbits (whose age varied between 7 and 9 weeks) divided into 5 lots of 3 animals. The average weight of these animals was 790 g. The results of this study show that the feces samples and the weight of young rabbits before administration of the treatment and the coprological examination (every 7 days for 4 weeks) show a fecal excretion reduction rate (FECRT) of 55.13% in the lot treated by sulfonamide. On the other hand, in animals received treatments extract of the leaves of Artemisia annua L., the average FECRT is evaluated at 69.64%, 79.22%, and 96.36% for respective doses of 400, 800 and 1200 mg/kg bodyweight and proves their anticoccidial effect. Furthermore, the variation in mean Eggs Per Gram (EPG) of coccidia and the average weekly weight gain (AWWG) of each lot were significant in the lots treated with hydro-ethanolic extract (P 0.05). The greatest reductions in oocystal excretion and weight gain obtained were those of lot 5, treated at 1200 mg/kg of hydro-ethanolic leaves extract of Artemisia annua L.

Artemisia annua: A New Version of a Traditional Tea under Randomized, Controlled Clinical Trial for the Treatment of Malaria

Introduction: The traditional antimalarial tea Artemisia annua, indicated for centuries in China to treat fevers, is again arousing interest for the treatment of malaria due to improvements attained in the plant composition by a few Institutions throughout the world, including the State University of Campinas (UNICAMP), Brazil, increasing its principal component by more than 100 times as from standard varieties, giving 1% in artemisinin and an expressive biomass yield such as 2 tons of dried leaves/hectare. Clinical trials carried out with this material in African countries have proven its therapeutic potential for a new generation of Artemisia tea in the treatment of falciparum malaria. In addition to artemisinin, recent studies have identified and quantified other compounds present in the crude extract and characterized their contributions to the anti-malarial efficacy, including their action against chloroquine-resistant strains. The majority of the clinical trials carried out with Artemisia tea in African countries have shown that the control of the parasitaemia is efficient in the initial treatment period, but few trials have followed the patients up to the 28th day. This first clinical trial carried out in Brazil with the A. annua infusion, after toxicological trials that defined the safety of this form of medication. Methods: The therapeutic efficacy of the tea was measured in patients with falciparum malaria over 28 days, comparing it with the current first-line treatment namely artemether-lumefantrine (Coartem?). The trial was carried out in controlled groups according to official protocol approved by the National Ethics in Research Committee (CONEP: 77/ 2011) and a rigorous control of the 17 patients with non-serious cases of falciparum malaria, recruited in the following three municipalities of the State of Pará, Brazil: Tucuruí, Goianésia do Pará and Anajás. The tea group received the infusion prepared in the proportion of 1.25 g of dry leaves of the variety CPQBA with 1% artemisinin for 250 mL of just boiled water, taken every 6 hours for 7 days, giving a total of approximately 175 mg of artemisinin, whilst the artemether-lumefantrine (Coartem?) group received a total of 525 mg of artemisinin equivalent to artemether. Results: The parasitaemia by the tea treatment became negative in the first days, even though it was administered with a dose that was one third of the recommended dose of artemisinin. However, as in the case artemisinin or artesunate monotherapies, 57.1% of the patients treated with the A. annua tea presented type I resistance, with a return of the parasitaemia around the 14th or 21st day. The other patients in the tea group showed type II/III resistance without manifestation of any serious signs or symptoms. In these cases, according to the protocol, the patients were redirected for treatment with artemether-lumefantrin (Coartem?) with subsequent negativity of the parisitaemia. Discussion: The fact that the efficacy of the tea with 1/3 of the dose of artemisinin was similar to that of the full dose of this medication infers that other compounds present in the crude extract, probably flavonoids, had contributed to the negativity of the parasitaemia at the start of the treatment. Considering that the positive control group, where the compound derived from artemisinin (artemether) was associated with another antimalarial agent (lumefantrine), presented excellent efficacy throughout the entire control of the cure, future trials with the A. annua tea should use the same strategy of association with another antimalarial agent, preferably from A. annua itself, in order to extend its therapeutic action during the whole control period. The Artemisia annua tea in the form standardized and used in this research, should not substitute the most efficient treatment, but could be considered as an emergency therapeutic resource in the first hours of symptoms as a function of its availability, anti-inflammatory action and lack of side effects. Other regimes and standardizations deserve investigation, mainly those with a high content of arteannuin B, as occurs in the initial cultivation phase.

The AaBBX21–AaHY5 module mediates light-regulated artemisinin biosynthesis in Artemisia annua L.

The sesquiterpene lactone artemisinin is an important anti-malarial component produced by the glandular secretory trichomes of sweet wormwood(Artemisia annua L.).Light was previously shown to promote artemisinin production,but the underlying regulatory mechanism remains elusive.In this study,we demonstrate that ELONGATED HYPOCOTYL5(HY5),a central transcription factor in the light signaling pathway,cannot promote artemisinin biosynthesis on its own,as the binding of AaHY5 to the promoters of artemisinin biosynthetic genes failed to activate their transcription.Transcriptome analysis and yeast two-hybrid screening revealed the B-box transcription factor AaBBX21 as a potential interactor with AaHY5.AaBBX21 showed a trichome-specific expression pattern.Additionally,the AaBBX21–AaHY5 complex cooperatively activated transcription from the promoters of the downstream genes AaGSW1,AaMYB108,and AaORA,encoding positive regulators of artemisinin biosynthesis.Moreover,AaHY5 and AaBBX21 physically interacted with the A.annua E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1(COP1).In the dark,AaCOP1 decreased the accumulation of AaHY5 and AaBBX21 and repressed the activation of genes downstream of the AaHY5–AaBBX21 complex,explaining the enhanced production of artemisinin upon light exposure.Our study provides insights into the central regulatory mechanism by which light governs terpenoid biosynthesis in the plant kingdom.

Graphene enhances artemisinin production in the traditional medicinal plant Artemisia annua via dynamic physiological processes and miRNA regulation

We investigated the effects of graphene on the model herb Artemisia annua,which is renowned for produc-ing artemisinin,a widely used pharmacological compound.Seedling growth and biomass were promoted when A.annua was cultivated with low concentrations of graphene,an effect which was attributed to a 1.4-fold increase in nitrogen uptake,a 15%–22%increase in chlorophyllfluorescence,and greater abun-dance of carbon cycling–related bacteria.Exposure to 10 or 20 mg/L graphene resulted in a�60%increase in H2O2,and graphene could act as a catalyst accelerator,leading to a 9-fold increase in catalase(CAT)ac-tivity in vitro and thereby maintaining reactive oxygen species(ROS)homeostasis.Importantly,graphene exposure led to an 80%increase in the density of glandular secreting trichomes(GSTs),in which artemisinin is biosynthesized and stored.This contributed to a 5%increase in artemisinin content inmature leaves.Inter-estingly,expression of miR828 was reduced by both graphene and H2O2 treatments,resulting in induction of its target gene AaMYB17,a positive regulator of GST initiation.Subsequent molecular and genetic assays showed that graphene-induced H2O2 inhibits micro-RNA(miRNA)biogenesis through Dicers and regulates the miR828–AaMYB17 module,thus affecting GST density.Our results suggest that graphene may contribute to yield improvement in A.annua via dynamic physiological processes together with miRNA regulation,and it may thus represent a new cultivation strategy for increasing yield capacity through nanobiotechnology.

青蒿苯丙氨酸解氨酶(AaPAL)基因家族鉴定及光调控分析

苯丙氨酸解氨酶是催化L-苯丙氨酸脱氨以进入苯丙烷途径第一步反应的酶,其对植物生长调节和抗逆反应都有着重要作用。从全基因组层次分析青蒿苯丙氨酸解氨酶基因家族的进化关系,可为相关基因功能研究提供线索。本研究利用HMMER和blastp从青蒿基因组中筛选苯丙氨酸解氨酶基因家族成员,并对其序列特征、进化关系以及表达模式等进行分析。最终从青蒿基因组中筛选到了6个苯丙氨酸解氨酶基因家族成员,其编码蛋白的氨基酸残基数量范围为446~716,蛋白分子量范围为49 559.81~77 621.57 kD,等电点(pI)范围为5.41~5.92,蛋白结构预测显示这些蛋白皆为亲水性蛋白;进化分析结果表明,6个AaPAL蛋白均与双子叶植物PAL蛋白的亲缘关系较近,同时这些蛋白序列具有保守的基序;通过对其顺式作用元件分析得知,苯丙氨酸解氨酶基因成员的启动子区域内含有较多的光响应、逆境胁迫响应、激素响应和生长发育相关的元件;青蒿的6个苯丙氨酸解氨酶基因在不同光处理下有着不同的表达方式;通过分析6个苯丙氨酸解氨酶基因的序列特征和表达模式,为深入研究该基因家族成员的功能及其对光调控下黄酮化合物生物合成调控机制提供参考。