Erigeron annuus Extract Improves DNCB-Induced Atopic Dermatitis in a Mouse Model via the Nrf2/HO-1 Pathway.

Atopic dermatitis (AD) is a persistent inflammatory skin condition resulting from an intricate interplay among genetic, immunological, and environmental factors. Erigeron annuus (EA), an annual winter plant belonging to the family Asteraceae, possesses anti-inflammatory, cytoprotective, and antioxidant activities. In this study, we hypothesized that Erigeron annuus extract (EAE) could be an effective agent for ameliorating AD-like symptoms. To confirm this hypothesis in vitro, we used H2O2-stimulated human keratinocytes (HaCaT cells) to demonstrate that pre-treatment with EAE protected against oxidative stress. HaCaT cells pretreated with EAE and stimulated with H2O2 showed decreased intracellular malondialdehyde content, increased superoxide dismutase activity, and reduced intracellular reactive oxygen species accumulation. To verify the in vivo hypothesis based on the intracellular results, an AD disease mouse model was induced with 1-chloro-2,4-dinitrobenzene (DNCB), and EAE was orally administered at a non-toxic concentration according to the toxicity evaluation results. The results showed that AD disease models in BALB/c mice exhibited reduced ear epidermal thickness, scratching behavior, and mast cell infiltration. In conclusion, our results indicate that EAE has the potential to improve AD by upregulating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway.

Genome-wide identification and characterization of SRLK gene family reveal their roles in self-incompatibility of Erigeron breviscapus.

Self-incompatibility (SI) is a reproductive protection mechanism that plants acquired during evolution to prevent self-recession. As the female determinant of SI specificity, SRK has been shown to be the only recognized gene on the stigma and plays important roles in SI response. Asteraceae is the largest family of dicotyledonous plants, many of which exhibit self-incompatibility. However, systematic studies on SRK gene family in Asteraceae are still limited due to lack of high-quality genomic data. In this study, we performed the first systematic genome-wide identification of S-locus receptor like kinases (SRLKs) in the self-incompatible Asteraceae species, Erigeron breviscapus, which is also a widely used perennial medicinal plant endemic to China.52 SRLK genes were identified in the E. breviscapus genome. Structural analysis revealed that the EbSRLK proteins in E. breviscapus are conserved. SRLK proteins from E. breviscapus and other SI plants are clustered into 7 clades, and the majority of the EbSRLK proteins are distributed in Clade I. Chromosomal and duplication analyses indicate that 65% of the EbSRLK genes belong to tandem repeats and could be divided into six tandem gene clusters. Gene expression patterns obtained in E. breviscapus multiple-tissue RNA-Seq data revealed differential temporal and spatial features of EbSRLK genes. Among these, two EbSRLK genes having high expression levels in tongue flowers were cloned. Subcellular localization assay demonstrated that both of their fused proteins are localized on the plasma membrane. All these results indicated that EbSRLK genes possibly involved in SI response in E. breviscapus. This comprehensive genome-wide study of the SRLK gene family in E. breviscapus provides valuable information for understanding the mechanism of SSI in Asteraceae.

Erigeron annuus Extract Alleviates Insulin Resistance via Regulating the Expression of Mitochondrial Damage and Endoplasmic Reticulum Stress-Related Genes.

Diabetes is a prevalent and debilitating metabolic disorder affecting a large population worldwide. The condition is characterized by insulin resistance and impaired function of pancreatic ß-cells, leading to elevated blood glucose levels. In this study, the antidiabetic effects of Erigeron annuus extract (EAE) on zebrafish with damaged pancreatic islets caused by insulin resistance were investigated. The study utilized the zebrafish model to monitor live pancreatic islets. RNA sequencing was also conducted to determine the mechanism by which EAE exerts its antidiabetic effect. The results showed that EAE was effective in recovering reduced islets in excess insulin-induced zebrafish. The effective concentration at 50% (EC50) of EAE was determined to be 0.54 µg/mL, while the lethal concentration at 50% (LC50) was calculated as 202.5 µg/mL. RNA sequencing indicated that the mode of action of EAE is related to its ability to induce mitochondrial damage and suppress endoplasmic reticulum stress. The findings of this study demonstrate the efficacy and therapeutic potential of EAE in treating insulin resistance in zebrafish. The results suggest that EAE may offer a promising approach for the management of diabetes by reducing mitochondrial damage and suppressing endoplasmic reticulum stress. Further research is required to establish the clinical application of EAE in diabetic patients.

Effects of breviscapine on cerebral ischemia-reperfusion injury and intestinal flora imbalance by regulating the TLR4/MyD88/NF-κB signaling pathway in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The plant Erigeron breviscapus (Vant.) Hand.-Mazz.,a Chinese herbal medicine with multiple pharmacological effects and clinical applications, has been traditionally used in the treatment of paralysis caused by stroke and joint pain from rheumatism by the Yi minority people of Southwest China for generations.However, its mechanism involves many factors and has not been fully clarified. AIM OF THE STUDY: Taking intestinal flora as the target, the protective effect of extract(breviscapine) of E. breviscapus on cerebral ischemia and its possible mechanism were discussed from the perspective of brain inflammatory pathway and intestinal CYP3A4, which depends on intestinal flora. MATERIALS AND METHODS: In this study, we first verified the binding ability between major active ingredient of Erigeron breviscapus and the core target TLR4 protein by molecular docking using Vina software.We established a rat model of cerebral ischemia-reperfusion injury in vivo.The neurological function of rats was scored by Bederson score table, the cerebral infarction volume was detected by TTC staining, and the serum NSE level was detected by ELASA. 16S rRNA sequencing was used to detect the intestinal flora of rats in each group.The expression levels of cerebral TLR4/MyD88/NF-κB and CYP3A4 mRNA and protein in different intestinal segments were detected by qRT-PCR and Western blot. RESULTS: Compared with the model group, the neurological injury score, infarct volume and serum NSE concentration of breviscapine low, medium and high dose groups and nimodipine groups decreased significantly. Meanwhile, breviscapine could significantly reduce the expression level of the TLR4/MyD88/NF-κB in brain tissue and CYP3A4 in different intestinal segments of rats with cerebral ischemia-reperfusion injury. In addition, breviscapine also significantly ameliorated intestinal flora dysbiosis of rats with cerebral ischemia-reperfusion injury. CONCLUSIONS: Breviscapine can protect rats from cerebral ischemia-reperfusion injury by regulating intestinal flora, inhibiting brain TLR4/MyD88/NF-κB inflammatory pathway and intestinal CYP3A4 expression.

Cytotoxic Effect of the Essential oils from Erigeron Canadensis L. on Human Cervical Cancer HeLa Cells in Vitro.

Erigeron Canadensis L. (E. canadensis) is a widely distributed invasive weed species in China. Potentially anti-cancer qualities may exist in its essential oils (EOs). The purpose of this study was to analyze the components of the EOs of E. canadensis and their effects on the normal liver cell lines L02 and the human cervical cancer cell lines HeLa. The EOs from the upper region of E. canadensis were prepared, its components were identified by GC/MS. Cell viability, cell morphology observation, AO/EB dual fluorescence staining assay, flow cytometry, mitochondrial membrane potential, western blot, caspase inhibitor test, and oxidative stress tests were used to investigate the impact of the EOs on HeLa cells. Network pharmacological analysis was employed to study the potential mechanism of the EOs in the treatment of cervical cancer. According to the findings, the EOs had 21 chemical components, of which limonene made up 65.68 %. After being exposed to the EOs, the cell viability of HeLa and L02 dramatically declined. The inhibition of EOs was more effective than that of limonene when used in an amount equivalent to that in the EOs. L02 cells were less susceptible to the cytotoxicity of EOs than HeLa cells were. Furthermore, EOs altered the cell cycle in HeLa cells and caused oxidative stress and apoptosis. Compared with the control group, the reactive oxygen species (ROS) levels increased in HeLa cells at first and then decreased, total superoxide dismutase (SOD) and catalase (CAT) activities in HeLa cells significantly decreased. G1 phase cells decreased whereas G2/M phase cells increased. The rate of apoptosis rose. Reduced mitochondrial membrane potential and Caspase-3, -9, and -12 protein expression were both observed. Nerolidol, dextroparaffinone, and α-pinene were shown to be the primary components for the suppression of HeLa cells, according to the results of the prediction of pharmacologic targets. In conclusion, findings of this study indicated the EOs may have the potential to curb the growth of cervical cancer cells. Further research is needed to explore the in vivo effect of EOs.

Sulfur fertiliser enhancement of Erigeron breviscapus (Asteraceae) quality by improving plant physiological responses and reducing soil cadmium bioavailability.

Erigeron breviscapus (Vant.) Hand.-Mazz. is an important medicinal plant; however, its quality is severely diminished by cadmium (Cd) pollution. Sulfur fertilisation can improve the production and application of E. breviscapus. This study examined Cd stress alleviation in the soil-plant system and determined the plant growth response after the application of sulfur fertiliser. The soil Cd concentration used in the treatments was 100 g·kg-1, and the sulfur fertiliser application rates were 0.1, 0.2, and 0.3 g·kg-1. Using pot experiments, we explored the impacts of high, medium, and low amounts of sulfur fertiliser on Cd accumulation and the quality and activity of E. breviscapus. The results showed that the application of sulfur fertiliser promoted Cd transformation to residual Cd under oxidation conditions, reducing Cd accumulation in E. breviscapus. Throughout the growth period, the application of sulfur fertiliser increased the soluble protein content and antioxidant enzyme activity, which alleviated Cd toxicity. The net photosynthetic rate, transpiration rate, intercellular CO2 concentration, chlorophyll level, and leaf width increased significantly. The biomass content of E. breviscapus also increased. Sulfur fertiliser improves the quality of herbaceous medicinal plants by reducing Cd accumulation and increasing scutellarin, chlorogenic, isochlorogenic acid B, and isochlorogenic acid C contents. A reasonable application of sulfur fertiliser is essential for improving E. breviscapus quality. This study provides a new method to reduce the ecological risk of planting herbaceous medicinal plants in Cd-contaminated soil.

Erigeron annuus flower absolute prolongs botulinum neurotoxin A-induced muscle paralysis and inhibits neurotransmitter release-linked responses.

This study aimed to determine the effects of Erigeron annuus (L.) Pers. (EAP) flower absolute (EAPFAb) on neurotransmitter release-blocking events and muscle paralysis induced by botulinum neurotoxin type A (BoNT/A). For this study, EAPFAb was extracted from EAP flowers by solvent extraction and its composition was determined by GC/MS. Neurotransmitter release and SNARE protein expression were examined in PC12 cells by ELISA and immunoblotting. Rat hind limbs were tested for muscle paralysis. EAPFAb contained 23 components and prolonged the duration of BoNT/A-induced rat hind limb muscle paralysis. EAPFAb reduced neurotransmitter release induced by elevated extracellular potassium levels and attenuated SNARE protein expression in PC12 cells. These findings demonstrate that EAPFAb prolongs BoNT/A-induced muscle paralysis action, probably by inhibiting releases of neurotransmitters that are regulated by SNARE proteins in neural cells. EAPFAb may be a promising material for prolonging BoNT/A action.

Comparative genomics reveal the convergent evolution of CYP82D and CYP706X members related to flavone biosynthesis in Lamiaceae and Asteraceae.

Distant species producing the same secondary metabolites is an interesting and common phenomenon in nature. A classic example of this is scutellarein whose derivatives have been used clinically for more than 30 years. Scutellarein occurs in significant amounts in species of two different orders, Scutellaria baicalensis and Erigeron breviscapus, which diverged more than 100 million years ago. Here, according to the genome-wide selection and functional identification of 39 CYP450 genes from various angiosperms, we confirmed that only seven Scutellaria-specific CYP82D genes and one Erigeron CYP706X gene could perform the catalytic activity of flavone 6-hydroxylase (F6H), suggesting that the convergent evolution of scutellarein production in these two distant species was caused by two independently evolved CYP450 families. We also identified seven Scutellaria-specific CYP82D genes encoding flavone 8-hydroxylase (F8H). The evolutionary patterns of CYP82 and CYP706 families via kingdom-wide comparative genomics highlighted the evolutionary diversity of CYP82D and the specificity of CYP706X in angiosperms. Multi-collinearity and phylogenetic analysis of CYP82D in Scutellaria confirmed that the function of F6H evolved from F8H. Furthermore, the SbaiCYP82D1A319D , EbreCYP706XR130A , EbreCYP706XF312D and EbreCYP706XA318D mutants can significantly decrease the catalytic activity of F6H, revealing the contribution of crucial F6H amino acids to the scutellarein biosynthesis of distant species. This study provides important insights into the multi-origin evolution of the same secondary metabolite biosynthesis in the plant kingdom.

Phytosynthesis of Palladium Nanoclusters: An Efficient Nanozyme for Ultrasensitive and Selective Detection of Reactive Oxygen Species.

Hydrogen peroxide is a low-reactivity reactive oxygen species (ROS); however, it can easily penetrate cell membranes and produce highly reactive hydroxyl radical species through Fenton's reaction. Its presence in abnormal amounts can lead to serious diseases in humans. Although the development of a simple, ultrasensitive, and selective method for H2O2 detection is crucial, this remains a strategic challenge. The peroxidase mimetic activity of palladium nanoclusters (PdNCs) has not previously been evaluated. In this study, we developed an ultrasensitive and selective colorimetric detection method for H2O2 using PdNCs. An unprecedented eco-friendly, cost-effective, and facile biological method was developed for the synthesis of PdNCs. This is the first report of the biosynthesis of PdNCs. The synthesized nanoclusters had a significantly narrow size distribution profile and high stability. The nanoclusters were demonstrated to possess a peroxidase mimetic activity that could oxidize peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB). Various interfering substances in serum (100 µM phenylalanine, cysteine, tryptophan, arginine, glucose, urea, Na+, Fe2+, PO43-, Mn+2, Ca2+, Mg2+, Zn2+, NH4+, and K+) were included to evaluate the selectivity of the assay, and oxidation of TMB occurred only in the presence of H2O2. Therefore, PdNCs show an efficient nanozyme for the peroxidase mimetic activity. The assay produced a sufficient signal at the ultralow concentration of 0.0625 µM H2O2. This colorimetric assay provides a real-time, rapid, and easy-to-use platform for the detection of H2O2 for clinical purposes.

Effect of Shoot Cutting on Trace Metal Concentration in Leaves and Capitula of Potential Phytoaccumulator, Invasive Erigeron annuus (Asteraceae).

The effect of shoot cutting was tested on cadmium, lead and zinc concentration in leaves and capitula of Erigeron annuus, an invasive species, which is considered as a potential phytoremediator. Plant material and soil were collected in the city center of Kraków, southern Poland, considered as one of the most contaminated cities in Europe. We proved that the concentration of zinc in leaves and capitula was higher after regrowth, concentration of cadmium was lower in capitula than in leaves, and the average value of bio-concentration factor for zinc and cadmium was less than 1, whereas for lead it was greater than 1 in both plant organs. Our results suggested that E. annuus can be potentially used for phytoremediation of lead and cutting the shoots can promote effectiveness of zinc removal from the contaminated soil.

Molecular cloning and expression analysis of SRLK1 gene in self-incompatible Asteraceae species Erigeron breviscapus.

Based on the transcriptome data, using RACE techniques, we cloned the full-length EbSRLK1 gene in a medicinal, self-incompatible Asteraceae species, Erigeron breviscapus. Bioinformatics approaches were used to analyze the DNA and protein sequences, physical and chemical properties, and domains of the encoded protein. The full-length EbSRLK1 cDNA is 2891 base pairs (bp) with an open reading frame (ORF) of 2634 bp, which encodes the EbSRLK1 protein with 878 amino acids and an estimated molecular weight of 98.13 kD. The EbSRLK1 protein has the characteristic domain structure of S-locus receptor-like protein kinases, which contains one transmembrane domain but lacks the signal peptide. Quantitative real-time PCR (qRT-PCR) analysis showed that the EbSRLK1 gene is lowly expressed in roots, stems and leaves, but highly expressed in flowers, especially in flowers one day prior to opening. Western blot analysis showed that the EbSRLK1 protein is expressed in stems, leaves, and flowers, but is almost undetectable in roots. The EbSRLK1 protein expression is induced in self-pollinated but not in cross-pollinated E. breviscapus flowers. Cloning and expression analysis of EbSRLK1 lay a solid foundation for elucidating the role of EbSRLK1 in regulating self-incompatibility in E. breviscapus.

Engineering yeast for the production of breviscapine by genomic analysis and synthetic biology approaches.

The flavonoid extract from Erigeron breviscapus, breviscapine, has increasingly been used to treat cardio- and cerebrovascular diseases in China for more than 30 years, and plant supply of E. breviscapus is becoming insufficient to satisfy the growing market demand. Here we report an alternative strategy for the supply of breviscapine by building a yeast cell factory using synthetic biology. We identify two key enzymes in the biosynthetic pathway (flavonoid-7-O-glucuronosyltransferase and flavone-6-hydroxylase) from E. breviscapus genome and engineer yeast to produce breviscapine from glucose. After metabolic engineering and optimization of fed-batch fermentation, scutellarin and apigenin-7-O-glucuronide, two major active ingredients of breviscapine, reach to 108 and 185 mg l-1, respectively. Our study not only introduces an alternative source of these valuable compounds, but also provides an example of integrating genomics and synthetic biology knowledge for metabolic engineering of natural compounds.

Coordinated role of soluble and cell wall bound phenols is a key feature of the metabolic adjustment in a mining woody fleabane (Dittrichia viscosa L.) population under semi-arid conditions.

Environmental contamination by hazardous heavy metals/metalloids (metal(loid)s) is growing worldwide. To restrict the migration of toxic contaminants, the establishment of a self-sustainable plant cover is required. Plant growth in multi-polluted soils is a challenging issue not only by metal(loid) toxicities, but also by the co-occurrence of other stressors. Dittrichia viscosa is a pioneer Mediterranean species able to thrive in metal(loid)-enriched tailings in semi-arid areas. The aim of the present work was to examine the metabolic adjustments involved in the acclimation responses of this plant to conditions prevailing in mine-tailings during Mediterranean spring and summer. For this purpose, fully-expanded leaves, and rhizosphere soil of both mining and non-mining populations of D. viscosa grown spontaneously in south-eastern Spain were sampled in two consecutive years. Quantitative analysis of >50 biochemical, physiological and edaphic parameters were performed, including nutrient status, metal(loid) contents, leaf redox components, primary and secondary metabolites, salicylic acid levels, and soil physicochemical properties. Results showed that mining plants exhibited high foliar Zn/Pb co-accumulation capacity, without substantially affecting their photosynthetic metabolism or nutritional status even in the driest summer period. The comparison of the antioxidative/oxidative profile between mining and non-mining D. viscosa populations revealed no major seasonal changes in the content of primary antioxidants (ascorbate and GSH), or in the levels of ROS. Multivariate analysis showed that phenylalanine ammonia-lyase (PAL) and peroxidase (PRX) activities and soluble and cell wall-bound phenols were potential biomarkers for discriminating between both populations. During the dry season, a marked enhancement in the activity of both PAL and soluble PRX resulted in both a drop in the accumulation of soluble phenols and an increase of the strong metal chelator caffeic acid in the cell-wall fraction, supporting the view that the plasticity of phenylpropanoid metabolism provide an effective way to counteract the effects of stress combinations.

Photoprotection of PSI by Far-Red Light Against the Fluctuating Light-Induced Photoinhibition in Arabidopsis thaliana and Field-Grown Plants.

It has been reported that PSI photoinhibition is induced even in wild-type plants of Arabidopsis thaliana, rice and other species by exposure of leaves to fluctuating light (FL) for a few hours. Because plants are exposed to FL in nature, they must possess protective mechanisms against the FL-induced photodamage. Here, using A. thaliana grown at various irradiances, we examined PSI photoprotection by far-red (FR) light at intensities comparable with those observed in nature. Dark-treated leaves were illuminated by red FL alternating high/low light at 1,200/30 µmol m-2 s-1 for 800 ms/10 s. By this FL treatment without FR light for 120 min, the level of photo-oxidizable P700 was decreased by 30% even in the plants grown at high irradiances. The addition of continuous FR light during the FL suppressed this damage almost completely. With FR light, P700 was kept in a more oxidized state in both low- and high-light phases. The protective effect of FR light was diminished more in mutants of the NADH dehydrogenase-like complex (NDH)-mediated cyclic electron flow around PSI (CEF-PSI) than in the PGR5 (proton gradient regulation 5)-mediated CEF-PSI, indicating that the NDH-mediated CEF-PSI would be a major contributor to PSI photoprotection in the presence of FR light. We also confirmed that PSI photoinhibition decreased with the increase in growth irradiance in A. thaliana and field-grown plants, and that this PSI photodamage was largely suppressed by addition of FR light. These results clearly indicate that the most effective PSI protection is realized in the presence of FR light.

Photo-induced biosynthesis of silver nanoparticles using aqueous extract of Erigeron bonariensis and its catalytic activity against Acridine Orange.

The green synthesis of silver nanoparticles (AgNPs) has reduced the pollution load in the environment to a greater extent by avoiding the use of hazardous chemicals. In the present work we have developed an ecofriendly and zero cost approach for the green synthesis of more stable and spherical AgNPs using aqueous extract of Erigeron bonariensis (AEE) which act as both reducing and stabilizing agent. The reaction of AEE and AgNO3 was carried out in direct sunlight for the instant biosynthesis of AgNPs within minutes. The biosynthesis was monitored by UV-vis spectroscopy which exhibited a sharp SPR band at 442 nm and 435 nm after 5 and 35 min of sunlight exposure. The optimum conditions for biosynthesis of AgNPs were found to be 2.5mM AgNO3 concentration, 1.5% (v/v) of AEE inoculum dose and 35 min of sunlight exposure. Presence of spherical AgNPs with average size 13 nm was confirmed by SEM and TEM analysis. The XRD and SAED analysis confirmed the crystalline nature of the AgNPs where the Bragg's diffraction pattern at (111), (200), (220) and (311) corresponded to face centered cubic crystal lattice of metallic silver. The average roughness of the synthesized AgNPs was 3.21 nm which was confirmed by AFM analysis. FTIR analysis was recorded between 4000 and 400 cm(-1) which confirmed the involvement of various functional groups in the synthesis of AgNPs. The AgNPs thus obtained showed catalytic activity towards degradation of Acridine Orange (AO) without involvement of any hazardous reducing agent. The concentration dependent catalytic activity of the synthesized AgNPs was also monitored using 1, 2 and 3 mL of silver colloids and was found that the degradation of AO followed pseudo first-order kinetics.

Deep Sequencing Reveals the Effect of MeJA on Scutellarin Biosynthesis in Erigeron breviscapus.

BACKGROUND: Erigeron breviscapus, a well-known traditional Chinese medicinal herb, is broadly used in the treatment of cerebrovascular disease. Scutellarin, a kind of flavonoids, is considered as the material base of the pharmaceutical activities in E. breviscapus. The stable and high content of scutellarin is critical for the quality and efficiency of E. breviscapus in the clinical use. Therefore, understanding the molecular mechanism of scutellarin biosynthesis is crucial for metabolic engineering to increase the content of the active compound. However, there is virtually no study available yet concerning the genetic research of scutellarin biosynthesis in E. breviscapus. RESULTS: Using Illumina sequencing technology, we obtained over three billion bases of high-quality sequence data and conducted de novo assembly and annotation without prior genome information. A total of 182,527 unigenes (mean length = 738 bp) were found. 63,059 unigenes were functionally annotated with a cut-off E-value of 10(-5). Next, a total of 238 (200 up-regulated and 38 down-regulated genes) and 513 (375 up-regulated and 138 down-regulated genes) differentially expressed genes were identified at different time points after methyl jasmonate (MeJA) treatment, which fell into categories of 'metabolic process' and 'cellular process' using GO database, suggesting that MeJA-induced activities of signal pathway in plant mainly led to re-programming of metabolism and cell activity. In addition, 13 predicted genes that might participate in the metabolism of flavonoids were found by two co-expression analyses in E. breviscapus. CONCLUSIONS: Our study is the first to provide a transcriptome sequence resource for E. breviscapus plants after MeJA treatment and it reveals transcriptome re-programming upon elicitation. As the result, several putative unknown genes involved in the metabolism of flavonoids were predicted. These data provide a valuable resource for the genetic and genomic studies of special flavonoids metabolism and further metabolic engineering in E. breviscapus.

Phytoextraction of metals by Erigeron canadensis L. from fly ash landfill of power plant "Kolubara".

The objectives of this study were to determine the concentrations of Pb, Cd, As, Cr, Cu, Co, Ni, Zn, Ba, Fe, Al and Ag in Erigeron canadensis L. growing on fly ash landfill of power plant "Kolubara", Serbia. The content of each element was determined in every part of plant separately (root, stalk and inflorescence) and correlated with the content of elements in each phase of sequential extraction of fly ash. In order to ambiguously select the factors that are able to decidedly characterize the particular part of plant, principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed. The bioconcentration factors and translocation factors for each metal were calculated in order to determine the feasibility of the use of plant E. canadensis L. for phytoremediation purpose. There were strong positive correlations between metals in every part of plant samples, and metals from pseudo total form of sequential extraction indicate that the bioavailability of elements in fly ash is similarly correlated with total form. Retained Al, Fe, Cr and Co in the root indicate its suitability for phytostabilization. This plant takes up Cd and Zn from the soil (bioconcentration factors (BCFs) greater than 1), transporting them through the stalk into the inflorescence (translocation factors (TFs) higher than 1). Regarding its dominance in vegetation cover and abundance, E. canadensis L. can be considered adequate for phytoextraction of Cd and Zn from coal ash landfills at Kolubara.

Analysis of the transcriptome of Erigeron breviscapus uncovers putative scutellarin and chlorogenic acids biosynthetic genes and genetic markers.

BACKGROUND: Erigeron breviscapus (Vant.) Hand-Mazz. is a famous medicinal plant. Scutellarin and chlorogenic acids are the primary active components in this herb. However, the mechanisms of biosynthesis and regulation for scutellarin and chlorogenic acids in E. breviscapus are considerably unknown. In addition, genomic information of this herb is also unavailable. PRINCIPAL FINDINGS: Using Illumina sequencing on GAIIx platform, a total of 64,605,972 raw sequencing reads were generated and assembled into 73,092 non-redundant unigenes. Among them, 44,855 unigenes (61.37%) were annotated in the public databases Nr, Swiss-Prot, KEGG, and COG. The transcripts encoding the known enzymes involved in flavonoids and in chlorogenic acids biosynthesis were discovered in the Illumina dataset. Three candidate cytochrome P450 genes were discovered which might encode flavone 6-hydroase converting apigenin to scutellarein. Furthermore, 4 unigenes encoding the homologues of maize P1 (R2R3-MYB transcription factors) were defined, which might regulate the biosynthesis of scutellarin. Additionally, a total of 11,077 simple sequence repeat (SSR) were identified from 9,255 unigenes. Of SSRs, tri-nucleotide motifs were the most abundant motif. Thirty-six primer pairs for SSRs were randomly selected for validation of the amplification and polymorphism. The result revealed that 34 (94.40%) primer pairs were successfully amplified and 19 (52.78%) primer pairs exhibited polymorphisms. CONCLUSION: Using next generation sequencing (NGS) technology, this study firstly provides abundant genomic data for E. breviscapus. The candidate genes involved in the biosynthesis and transcriptional regulation of scutellarin and chlorogenic acids were obtained in this study. Additionally, a plenty of genetic makers were generated by identification of SSRs, which is a powerful tool for molecular breeding and genetics applications in this herb.

[Relationship between expression of chalcone synthase gene (CHS) and scutellarin content in Erigeron breviscapus].

OBJECTIVE: Scutellarin from Erigeron breviscapus is a flavonoid with remarkable pharmacological activity, whose route of biosynthesis is still fully clear. Chalcone synthase (CHS) is the key enzyme regulating flavonoids biosynthesis, and the aim of this study is to explain the relationship between patterns of the gene expression and scutellarin content through studying CHS gene expression patterns combined with scutellarin content in various parts of E. breviscapus. METHOD: Through RT-PCR and RACE, the full length of CHS was cloned and analyzed by fluorescent quantitative PCR. The scutellarin content in tissues was analyzed by HPLC. RESULT: The full-length gene sequence was 1 270 bp, encoding 405 amino acids. Software analysis found that the DNA sequence was 80% similarity with Compositae plant homeo-box gene. Fluorescence quantitative analysis showed that CHS had the highest expression level in leaves, far higher than that in root, stem and flower. HPLC analysis showed that the scutellarin was the highest in leaves, followed by the flowers and stems, scutellarin was not detected in root. CONCLUSION: Correlation analysis showed that CHS expression amount and scutellarin content in different parts of E. breviscapus is positive correlation (r = 0.761, P < 0.05), it suggests that CHS gene expression level has important effect on biosynthesis of scutellarin.

[Genetic diversity and group consistency of breeding strains of Erigeron breviscapus determined by AFLP marker].

OBJECTIVE: To analyze the genetic diversity and breeding strains of the E. breviscapus germplasms, in order to provide theoretical information for Erigeron breviscapus breeding. METHOD: The genetic diversity and genetic structure were assayed to six germplasm resource of E. breviscapus which collected from Yunnna with 11 pairs primers and AFLP molecular marker. RESULT: Six hundred and four amplification bands among 636 DNA bands were from six accession of E. breviscapus, which are about 82.40% of total bands. The six germplasms could be divided into three group at the 0. 706 similarity coefficient level. The first category include QS-1, QS-2 and Dali, Shilin, Kunming population. The second category included wild population of Qiubei. The third category included several sample from different district. The mean genetic similarity coefficient of QS-1 and QS-2 was bigger, genetic similarity coefficient range was smaller, hereditary character was more stable. Molecular system clustering analysis showed that the geographical origin of the same part had relative polymerization phenomenon and its genetic relationship was close. Qiubei was a single group possibly relating to the specific genetic basis. CONCLUSION: The analysis of genetic diversity of E. breviscapus by AFLP marker is reliable. The systematic E. breviscapus breeding is feasible.