Synthesis Of Chlorogenic Acid Research Articles

Unraveling the Distribution, Metabolization, and Catabolism of Foliar Sprayed Carbon Dots in Maize and Effect on Soil Environment.

The enormous potential of carbon dots (CDs) in agriculture has been widely reported, whereas their accurate distribution, transformation, and metabolic fate and potential soil health effects are not clearly understood. Herein, 13C-labeled CDs (13C-CDs) were sprayed on maize leaf, accumulated in all tissues, and promoted photosynthesis. Specifically, 13C-CDs were internalized to participate in the synthesis of glucose, sucrose, citric acid, glyoxylate, and chlorogenic acid, promoting tricarboxylic acid cycle (TCA) and phenylalanine metabolism. Additionally, the catabolism of 13C-CDs in vivo was mainly mediated by O2•- produced by oxidative stress. 13C-CDs did not have an obvious impact on the soil environment at the overall level. The detection of 13C signals in soil fauna suggested 13C-CDs in soil food chain transmission. This study systematically described the exact fate of CDs in plants and potential soil ecological risks and provided a more comprehensive analysis and support for the potential advantages of CDs in agricultural application.

Multiplex Approach of Metabolomic and Transcriptomic Reveals the Biosynthetic Mechanism of Light-induced Flavonoids and CGA in Chrysanthemum

Light, as an important environmental signal for plant growth and development, has been reported to be involved in the metabolism of phenylpropane. However, the light-induced phenylpropane biosynthesis mechanism in Chrysanthemum morifolium Ramat., especially flavonoid and chlorogenic acid (CGA) metabolism, is not clear. In this study, we found the flower phenotype of chrysanthemum 'HangBaiJu' became lighter after bagging. Besides, the content of main active ingredients, such as Luteolin-7-O-glucoside (Lu-7-O-G), Diosmetin-7-O-glucoside (Di-7-O-G), Apigenin-7-O-glucoside, Apigenin and CGA, was significantly decreased. Comparative transcriptome analysis (pairwise comparison, WGCNA, and k-means clustering) revealed that the expression of structural genes, such as CmPAL, CmCHS1/2, CmF3’H, CmFNS and CmHQT, was notably declined under dark conditions. Meanwhile, ‘bridge proteins’ CmMYB3/6/16 and signal transduction factors CmBBX20/22.2/22.3 were identified as key regulatory factors in the light-mediated synthesis of flavonoids and CGA in chrysanthemum. Among that, CmBBX20 could promote the accumulation of flavonoids and CGA by directly binding to the G-box core sequences of downstream target genes based on transient overexpression and Y1H assays. Overall, the preliminary molecular mechanism of BBXs and MYBs coordinately regulating the accumulation of flavonoids and CGA in chrysanthemum under different light inductions has been clarified, which provided a theoretical basis for the molecular breeding and quality improvement of chrysanthemum.

Full-length transcriptome sequencing and transgenic tobacco revealed the key genes in the chlorogenic acid synthesis pathway of Sambucus chinensis L.

Chlorogenic acid is a key chemical in antioxidation and antisepsis. Sambucus chinensis L. is a herbaceous plant rich in chlorogenic acid and a potential genetic resource for breeding high-chlorogenic acid plants. However, there are few studies on the synthesis pathway of chlorogenic acid in S. chinensis. Our study found chlorogenic acid accumulation in S. chinensis to be organ-specific, higher in leaves and buds but lower in roots, stems and fruits. A total number of 546,844 CCS (Circular Consensus Sequence), including 402,767 full-length non-chimeric (FLNC) and 39 annotated sequences related to the synthesis of chlorogenic acid, was obtained by single-molecule real-time sequencing technology (SMRT). qRT-PCR showed that a number of key genes involved in chlorogenic acid synthesis were differentially expressed in various tissues of S. chinensis. Transgenic tobacco revealed that ectopic expression of the HCT homologous gene HCT-45178 increased the content of chlorogenic acid. Our results should be the first report of full-length transcriptome data of S. chinensis, which help to understand the basis of chlorogenic acid synthesis and provide a novel strategy for breeding tobacco cultivars with higher levels of chlorogenic acid. This article is protected by copyright. All rights reserved.

Identification and expression analysis of chlorogenic acid biosynthesis key gene PpHCT in peach

Shikimic acid/quinic acid hydroxy cinnamyl transferase (HCT) is one of the key enzymes in the phenylpropanoid pathway. However, the role of the HCT gene in chlorogenic acid (CGA) biosynthesis in peach fruit remains unclear. For this, we identified the accumulation pattern of CGA in four peach cultivars, cloned and characterized 11 PpHCT gene members, and further analyzed the expression patterns of these PpHCT genes during fruit development. The contents of CGAs in the four peach cultivars all exhibited a trend of increasing and then decreasing during the fruit growth and development. Moreover, the contents of CGAs in the peel and flesh were tissue-specific. Gene structure analysis indicated that the PpHCT genes were highly conserved, containing two exons and one intron. The protein structure analysis demonstrated that the PpHCT proteins contained two conserved motifs (HXXXD, DFGWG) and a transferase domain (PF02458), which belonged to the BAHD acyltransferase family. The cis-acting element analysis suggested that the promoters of PpHCT genes contained many light-related, hormone-related, stress-related, tissue-specific, and circadian-related elements, and they could participate in a variety of biological processes. Phylogenetic analysis showed that the HCT proteins of peach were closely related to the HCT proteins of plum and had a close evolutionary relationship. The qRT-PCR analysis indicated that the expression levels of PpHCT1 and PpHCT2 showed an opposite trend to the accumulation of CGA, whereas the expression levels of PpHCT4, PpHCT5, PpHCT7, PpHCT8, and PpHCT11 demonstrated the same trend as CGA accumulation. It was worth noting that only PpHCT4 and PpHCT5 were highly expressed in the two high-CGA cultivars but showed low levels of expression in the two low-CGA cultivars. Therefore, it was hypothesized that these two genes might be key genes to the synthesis of CGA in peach fruit. Those findings provide a theoretical basis for further study on the biological functions of the HCT gene and help to reveal the molecular mechanism of CGA.

Transcription factor NtWRKY33a modulates the biosynthesis of polyphenols by targeting NtMYB4 and NtHCT genes in tobacco

There are abundant polyphenols in tobacco leaves mainly including chlorogenic acid (CGA), rutin, and scopoletin, which not only influence plant growth, development, and environmental adaptation, but also have a great impact on the industrial utilization of tobacco leaves. Few transcription factors regulating the biosynthesis of polyphenols have been identified in tobacco so far. In this study, two NtWRKY33 genes were identified from N. tabacum genome. NtWRKY33a showed higher transcriptional activity than NtWRKY33b, and encoded a nuclear localized protein. Overexpression and knock-out of NtWRKY33a gene revealed that NtWRKY33a inhibited the accumulation of rutin, scopoletin, and total polyphenols, but meanwhile promoted the biosynthesis of CGA. Chromatin immunoprecipitation and Dual-Luc assays indicated that NtWRKY33a could directly bind to the promoters of NtMYB4 and NtHCT, and thus induced the transcription of these two genes. The contents of polyphenols in ntwrky33a, ntmy4, and ntwrky33a/ntmyb4 mutants further confirmed that the repression of NtWRKY33a on the biosynthesis of rutin, scopoletin, and total polyphenols depends on the activity of NtMYB4. Moreover, the promotion of NtHCT by NtWRKY33a modulates the distribution of metabolism flux into the synthesis of CGA. Ectopic expression of NtWRKY33a inhibit the expression of NtSAUR14, NtSAUR59, NtSAUR66, NtIAA4, NtIAA17, and NtIAA19 genes, indicating that NtWRKY33a might be involved in the regulation of plant auxin response. Our study revealed new functions of NtWRKY33a in regulating the synthesis of polyphenols, and provided a promising target for manipulating polyphenols contents in tobacco.

Heat Shock Treatment Promoted Callus Formation on Postharvest Sweet Potato by Adjusting Active Oxygen and Phenylpropanoid Metabolism

This study aimed to investigate that rapid high-temperature treatment (RHT) at an appropriate temperature could accelerate callus formation by effectively promoting the necessary metabolic pathways in sweet potato callus. In this study, the callus of sweet potato was treated with heat shock at 50, 65, and 80 °C for 15 min. The callus formation was observed within 1, 3, and 5 days, and the accumulation of intermediates in the metabolism of phenylpropane and reactive oxygen species and changes in enzyme activities were determined. The results showed that appropriate RHT treatment at 65 °C stimulated the metabolism of reactive oxygen species at the injury site of sweet potato on the first day, and maintained a high level of reactive oxygen species production and scavenging within 5 days. The higher level of reactive oxygen species stimulated the phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase and cinnamate-4-hydroxylase activities of the phenylpropane metabolic pathway, and promoted the rapid synthesis of chlorogenic acid, p-coumaric acid, rutin, and caffeic acid at the injury site, which stacked to form callus. By Pearson’s correlation analysis, catalase (CAT), PAL, and chlorogenic acid content were found to be strongly positively correlated with changes in all metabolites and enzymatic activities. Our results indicated that appropriate high-temperature rapid treatment could promote sweet potato callus by inducing reactive oxygen species and phenylpropane metabolism; moreover, CAT, PAL, and chlorogenic acid were key factors in promoting two metabolic pathways in sweet potato callus.

PbHCT4 regulates growth through affecting chlorogenic acid (CGA) content in pear

• The chlorogenic acid contents between dwarf and standard pear trees were different significantly. • Low concentration of chlorogenic acid promoted the growth of pear, while high concentration of chlorogenic acid inhibited the growth. • PbHCT4 gene changed plant height by regulating chlorogenic acid content and auxin transport. Due to its many advantages, dwarfing and dense planting is an important mode of modern fruit tree cultivation. Phenols have been reported to play various roles in the growth regulation of plants. Here, to verify the correlation between chlorogenic acid (CGA) and the growth of pear trees, the CGA content of standard and dwarf pear trees as well as the effect of exogenous CGA upon their growth, were investigated and analyzed. Our results showed that the CGA content of two dwarf-type varieties, ‘Zhongai 1′ (Z1) and ‘PY-9′ was significantly higher than that of two standard-type Pyrus betulaefolia (P1 and P2). Whereas a low concentration of CGA accelerated pear tree growth, and a high concentration of CGA could inhibit it. Then, through an expression pattern analysis of four CGA synthesis-related genes, PbHCT1–4 , we found that the dwarf phenotype might be related to the expression levels of PbHCTs , especially to those of PbHCT2–4 in shoots. Of these, PbHCT4 was selected as a candidate gene and transformed into tobacco. When compared with control tobacco plants, the transgenic plants attained taller heights and had a greater CGA content, as well as increased expression levels of the auxin efflux carrier gene, NbPIN. Accordingly, we speculate the PbHCT4 gene promotes the synthesis of CGA, which influences plant growth by mediating auxin's transport in pear trees. These findings are helpful for advancing analysis of dwarf mechanism, which is critical for the breeding of new dwarf pear varieties.

Genome-Wide Identification and Expression Analyses of the 4-Coumarate: CoA Ligase (4CL) Gene Family in Eucommia ulmoides

4-Coumarate: coenzyme A ligase (4CL) is an important rate-limiting enzyme in phenylpropanoid metabolism that plays an important role in the biosynthesis of phenylpropanoid, flavonoid, lignin and other secondary metabolites in plants. However, the family members and functions have not been identified in Eucommia ulmoides (E. ulmoides). In this study, 35 Eu4CLs were identified in the E. ulmoides genome, and the gene structure, conserved domain, evolutionary relationship and expression pattern were comprehensively analyzed. The results show that 35 Eu4CLs were assembled into three subgroups according to the classification in Arabidopsis, where Eu4CLs in the same subgroup had similar gene structures and conserved protein motifs. Putative cis-element analysis of Eu4CL promoter regions uncovered numerous elements related to the response of stress and plant hormones. Expression patterns showed that Eu4CL4/5/13/34 expression levels were positively related to chlorogenic acid content in different periods, which indicate that the synthesis of chlorogenic acid in E. ulmoides was regulated by multiple genes, and the genes regulating the synthesis of chlorogenic acid in different tissues were different. In addition, nine selected Eu4CL genes showed different expression patterns under cold, WeJA (methyl jasmonate), and ethylene by quantitative reverse transcription-PCR (qRT-PCR) assay, suggesting that Eu4CL genes not only play an important role in the synthesis of chlorogenic acid, but also plays an important role in the process of biotic and abiotic stress. Taken together, these findings provide theoretical reference for further exploring the molecular characteristics and biological functions of Eu4CL genes.

Synthesis of chlorogenic acid and p-coumaroyl shikimate byexpressing shikimate gene modules in Escherichia coli.

Chlorogenic acid and p-coumaroyl shikimate are hydroxycinnamic acid derivatives. These compounds are nutraceutical supplements due to their biological activities including prevention of cardiovascular disease and cancers. These two compounds were synthesized in Escherichia coli through two-culture system using two mutants, which are biochemically interdependent. The aim of this work was to improve the titres of their production in a single E. coli mutant in which all necessary genes were introduced. This was done by testing various shikimate gene combinations to determine the optimal gene combination for the synthesis of chlorogenic acid and p-coumaroyl shikimate. A series of gene modules harbouring shikimate pathway genes were constructs. Six gene module constructs for chlorogenic acid synthesis and eight constructs for p-coumaric acid synthesis were tested in order to find the best one. Chlorogenic acid synthesis showed highest with the gene module construct containing ydiB, aroB, aroGf , ppsA and tktA. Using the E. coli strain, 109.7mgL-1 chlorogenic acid was synthesized. The best gene module construct for the p-coumaroyl shikimate synthesis contained aroD and aroGf . In addition, we used two E. coli deletion mutant strains (ΔaroK and ΔaroL) to increase the final titre. The E. coli ΔaroK mutant harbouring this gene module construct synthesized 713.4mgL-1 of p-coumaroyl shikimate. The chlorogenic acid synthesis using the current system was approximately 35.4% higher of the titre than titres obtained with an alternative method that depends on co-cultivation of two mutants. At the same time, production of p-coumaroyl shikimate increased 5.8 times. The current study's findings indicate that our selection of the shikimate gene module contributed to increases in the levels of the substrates and could be applied to synthesize other compounds whose synthesis requires intermediates of the shikimate pathway.

Spectral quality as an eliciting agent in the production of phenolic compounds in the callus of Hyptis marrubioides Epling

Hyptis marrubioides Epling is a species of the Brazilian cerrado traditionally used to treat gastrointestinal and cutaneous infections, pain, and cramps. The use of visible and ultraviolet C (UVC) radiation is a promising strategy to optimize the production of the bioactive metabolites. Therefore, the effect of the spectral quality of light on the production of metabolites was evaluated in H. marrubioides callus. The callus was inoculated on MS medium with 50% of the salt concentration containing 2 mg L-1 naphthaleneacetic acid (NAA) and 1 mg L-1 benzylaminopurine (BAP). The callus cultures were exposed for 20 days to the spectral qualities of white light, blue, red, and blue + red as well as to darkness. In addition, the callus cultivated under white light were exposed to UVC on the 21st day for 0, 30, 60, 120, and 240 seconds. The exposure of H. marrubioides callus to blue light negatively affects the synthesis of phenolic compounds. Red light stimulates the synthesis of caffeic acid and luteolin. Darkness was the best condition among those studied because it was associated with the increased accumulation of caffeic acid, chlorogenic acid, rosmarinic acid, and luteolin. The exposure of H. marrubioides callus cultivated under white light to UVC radiation promoted an increase in the synthesis of chlorogenic acid, ferulic acid, rosmarinic acid, and luteolin.

English

Chlorogenic acid (CGA), as a kind of depside in plants, has a variety of beneficial effects on human health, which also plays an important role in helping plants resist a variety of stresses. Therefore, the biosynthetic pathway of CGA has been studied in many plants, however, the synthesis of CGA has not been well elucidated in Camellia sinensis. In our research, different CGA levels were detected between triploid tea variety „Qianfu 4‟ and diplont tea variety „Qianmei 419‟ using HPLC and the CGA content in triploid Camellia sinensis was greater than that in diploid Camellia sinensis. Transcriptome sequencing for diploid and triploid Camellia sinensis was employed to explore genes associated with CGA biosynthesis. Finally, 154,097 unigenes were obtained in total, of which 891 may be related to the biosynthesis of CGA. Furthermore, differentially expressed genes (DEGs) were screened between diploid and triploid Camellia sinensis, 32 DEGs were discovered to be related to CGA biosynthesis, including sixteen phenylalanine ammonia lyase (PAL) genes, three 4-coumarate coenzyme A ligase (4CL) genes, nine cinnamate 4-Hydroxylase (C4H) genes, four Hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HQT/HCT), and two hundred and twenty-one TFs including eighty-eight ERFs, forty-one bZIPs, forty-two MYBs and fifty WRKYs, which may also play an important role in the biosynthesis of CGA. Our results will lay the foundation for further exploration of the biosynthesis of CGA and revealing the related regulatory network in Camellia sinensis. © 2021 Friends Science Publishers

Free Radical Scavenging and Anti-Inflammatory Activity of Chlorogenic Acid Mediated Silver Nanoparticle

Introduction: Nanotechnology is a field of research and innovation concerned with building 'things' - generally, materials and devices on the scale of atoms and molecules. It is a booming field of this 21st century. The role of nano technology is becoming very crucial in nearly every aspect of life ranging from cosmetics to advanced bio technological approaches. In recent years, silver nanoparticles have gained considerable attention in the field of medicine. The green synthesis of nanoparticles eliminates the generation and use of hazardous substances and thus sustains non toxicity. Chlorogenic acids are phenolic compounds formed by the esterification of cinnamic acids. They exhibit various pharmacological Properties. Our study deals with the green synthesis of Chlorogenic acid mediated silver nanoparticles and assessment of their anti inflammatory and antioxidant properties.
 
 Aim: The present study aims at assessing the antioxidant and anti inflammatory property of chlorogenic acid mediated silver nanoparticle and investigating the efficacy of Chlorogenic acid mediated Silver nanoparticle.
 Materials and Methods: The methodology includes Green synthesis of Chlorogenic acid mediated Silver nano particle synthesis followed by tests for Anti inflammation and Anti oxidation.
 Results: Both the Anti inflammatory and Anti oxidation activity of the chlorogenic acid mediated silver nanoparticle had shown a proportionate increase in activity with increasing concentration of the compound.
 Conclusion: Chlorogenic acid mediated silver nanoparticles have shown significant anti inflammatory and anti oxidation activity and they are considered as potent anti inflammatory and antioxidant agents.

The effect of cutting style on the biosynthesis of phenolics and cellular antioxidant capacity in wounded broccoli

To explore the effect of cutting style on the biosynthesis of phenolic compounds and cellular antioxidant capacity in wounded broccoli subjected to different cutting styles (heads, florets, 1/2 florets and shredded florets), the mechanism of the accumulation of phenolic compounds was investigated at the transcriptional level, and cellular antioxidant capacity was measured using a breast cancer cell MCF-7 culture model. The results indicated that the relative expression of the genes encoding phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase and 4-coumarin coenzyme A ligase, three enzymes involved in phenylpropanoid metabolism, was upregulated and that contributed to the synthesis of individual phenolic compounds, including catechin, hydroxybenzoic acid, chlorogenic acid, caffeic acid, sinapic acid, catechin gallate, rutin, cinnamic acid and quercetin. This research constructes the phenol synthesis pathway in wounded broccoli. Moreover, the relative expression of critical genes including superoxide dismutase, peroxidase, catalase, glutathione peroxidase and glutathione reductase involved in the metabolism of reactive oxygen species (ROS) increased, resulting in enhanced antioxidant capacity in wounded broccoli. Cell antioxidant capacity (CAA) of heads, florets, 1/2 florets and shredded florets increased by 52.7%, 59.2%, 64.8% and 86.5%, respectively, compared to whole broccoli. The enhancement of CAA was greater as the intensity of wounding increased, indicating that enhancement of antioxidant activity occurred at the cellular level. This research helps provide a reliable and persuasive theoretical basis for nutritional value assessment at the cellular level in wounded broccoli.

<i>In-Silico</i> Validation and Development of Chlorogenic Acid (CGA) Loaded Polymeric Nanoparticle for Targeting Neurodegenerative Disorders

Background: Recent decades witnessed a significant growth in terms of phytocompounds based therapeutics, extensively explored for almost all types of existing disorders. They have also been widely investigated in Neurodegenerative disorders (NDDs) and Chlorogenic acid (CGA), a polyphenolic compound having potential anti-inflammatory and anti-oxidative properties, emerged as a promising compound in ameliorating NDDs. Owing to its poor stability, bioavailability and release kinetics, CGA needed a suitable nanocarrier based pharmaceutical design for targeting NDDs. Objective: The current study is aimed at the in-silico validation of CGA as an effective therapeutic agent targeting various NDDs followed by the fabrication of polymeric nanoparticles-based carrier system to overcome its pharmacological limitations and improve its stability. Methods: A successful in-silico validation using molecular docking techniques along with synthesis of CGA loaded polymeric nanoparticles (CGA-NPs) by ionic gelation method was performed. The statistical optimisation of the developed CGA-NPs was done by Box Behnken method and then the optimized formulation of CGA-NPs was characterised using particle size analysis (PSA), Transmission electron microscopy (TEM), Fourier Transform Infrared spectroscopy (FTIR) along with in-vitro release kinetics analysis. Results & Conclusion: The results attained exhibited average particle size of 101.9 ± 1.5 nm, Polydispersibility (PDI) score of 0.065 and a ZP of −17.4 mV. On a similar note, TEM results showed a size range of CGA-NPs between 90 - 110 nm with a spherical shape of NPs. Also, the data from in-vitro release kinetics showed a sustained release of CGA from the NPs following the first-order kinetics suggesting the appropriate designing of nanoformulation.

Impact of novel SNPs identified in Cynara cardunculus genes on functionality of proteins regulating phenylpropanoid pathway and their association with biological activities

BackgroundCynara cardunculus L. offers a natural source of phenolic compounds with the predominant molecule being chlorogenic acid. Chlorogenic acid is gaining interest due to its involvement in various biological properties such as, antibacterial, antifungal, antioxidant, hepatoprotective, and anticarcinogenic activities.ResultsIn this work we screened a Cynara cardunculus collection for new allelic variants in key genes involved in the chlorogenic acid biosynthesis pathway. The target genes encode p-coumaroyl ester 3′-hydroxylase (C3′H) and hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase (HQT), both participating in the synthesis of chlorogenic acid. Using high-resolution melting, the C3′H gene proved to be highly conserved with only 4 haplotypes while, for HQT, 17 haplotypes were identified de novo. The putative influence of the identified polymorphisms in C3′H and HQT proteins was further evaluated using bioinformatics tools. We could identify some polymorphisms that may lead to protein conformational changes. Chlorogenic acid content, antioxidant and antithrombin activities were also evaluated in Cc leaf extracts and an association analysis was performed to assess a putative correlation between these traits and the identified polymorphisms.ConclusionIn this work we identified allelic variants with putative impact on C3′H and HQT proteins which are significantly associated with chlorogenic acid content and antioxidant activity. Further study of these alleles should be explored to assess putative relevance as genetic markers correlating with Cynara cardunculus biological properties with further confirmation by functional analysis.

Improved production of chlorogenic acid from cell suspension cultures of <i>Lonicera macranthoids</i>

Purpose: To evaluate the potential of Lonicera macranthoides Hand. -Mazz. Yulei1 suspension culture system for enhanced production of the main secondary metabolite, chlorogenic acid.Methods: The callus of L. macranthoides Hand.-Mazz. “Yulei1” was suspension cultured in B5 liquid medium supplemented with different plant growth regulators. Biomass accumulation was calculated by weight method and chlorogenic acid production was measured using high performance liquid chromatography (HPLC). HPLC was carried out on C18 analytical column at 35 °C and the detection wavelength was set at 324 nm.Results: The results showed that maximum accumulation of biomass and chlorogenic acid were achieved 15 days after culture growth. The optimized conditions for biomass accumulation and chlorogenic acid production were 50 g/L of inoculum on fresh weight basis, B5 medium supplemented with plant growth regulators, 30 - 40 g/L sucrose and initial medium pH of 5.5. Maximum accumulation of chlorogenic acid and biomass were observed when the culture medium was supplemented with 2.0 mg/L6-BA. Optimal accumulation of chlorogenic acid was observed using combination of hormones 2.0 mg/L 6-Benzyladenine (BA) + 0.5 mg/L2, 4-Dichlorophenoxyacetic acid (2,4-D), while optimal accumulation of biomass was observed with 2.0 mg/L 6-BA + 2.0 mg/L2, 4-D. In addition, phenylalanine also contributed to the synthesis of chlorogenic acid at a concentration > 50 mg/L.Conclusion: Cell suspension cultures of L. macranthoides Hand.-Mazz. “Yulei1” have successfully been established. The findings provide a potential basis for large scale production of chlorogenic acid using cell suspension cultures of L. macranthoides.Keywords: Lonicera macranthoides, Cell suspension culture, Chlorogenic acid, Phenylalanine, Optimization

Synthesis and Application of Novel 3D Magnetic Chlorogenic Acid Imprinted Polymers Based on a Graphene-Carbon Nanotube Composite.

A novel three-dimensional (3D) magnetic chlorogenic acid (CGA) imprinted polymer (MMIP) was prepared with novel carbon hybrid nanocomposite as the carrier, chlorogenic acid as the template molecule, and methacrylic acid as the functional monomer. The 3D MMIPs were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, vibrating sample magnetometer, and UV spectrometry in detail. The results showed that the imprinted layer was attached successfully on the surface of a 3D magnetic carbon hybrid nanocomposite. The adsorption performance of the 3D MMIPs was investigated, and the results showed that the 3D MMIPs exhibited high adsorption capacity and fast adsorption rate toward CGA with a maximum adsorption capacity of 10.88 mg g(-1). The extraction conditions involving washing solvent, the pH of eluent solvent, elution volume, and desorption time were also investigated in detail. Combined with high-performance liquid chromatography, the 3D MMIPs have been applied to successfully extract CGA from Eucommia leaf extract samples.

Trends in the content of chemical elements in leaves of Lonicers Caerulea (Caprifoliaceae) in connection with their secondary metabolism in the natural populations of the Altai Mountains

SRXRF is used to study trends in the content of macro- and microelements in the assimilation organs of blue honeysuckle in geologically contrasting sectors of the Altai Mountains. The accumulation of flavonoids in the organs of L. caerulea is influenced primarily by Cu, Ni, and Zn, as these trace elements are selectively concentrated by this species. Intensified synthesis of chlorogenic acid and its derivatives is due to a considerable increase in Fe concentration in plant leaves.

Synthesis and regulation of chlorogenic acid in potato: Rerouting phenylpropanoid flux in HQT-silenced lines.

Chlorogenic acid (CGA) is the major phenolic sink in potato tubers and can constitute over 90% of total phenylpropanoids. The regulation of CGA biosynthesis in potato and the role of the CGA biosynthetic gene hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (HQT) was characterized. A sucrose induced accumulation of CGA correlated with the increased expression of phenylalanine ammonia-lyase (PAL) rather than HQT. Transient expression of the potato MYB transcription factor StAN1 (anthocyanin 1) in tobacco increased CGA. RNAi suppression of HQT resulted in over a 90% reduction in CGA and resulted in early flowering. The reduction in total phenolics and antioxidant capacity was less than the reduction in CGA, suggesting flux was rerouted into other phenylpropanoids. Network analysis showed distinct patterns in different organs, with anthocyanins and phenolic acids showing negative correlations in leaves and flowers and positive in tubers. Some flavonols increased in flowers, but not in leaves or tubers. Anthocyanins increased in flowers and showed a trend to increase in leaves, but not tubers. HQT suppression increased biosynthesis of caffeoyl polyamines, some of which are not previously reported in potato. Decreased PAL expression and enzyme activity was observed in HQT suppressed lines, suggesting the existence of a regulatory loop between CGA and PAL. Electrophysiology detected no effect of CGA suppression on potato psyllid feeding. Collectively, this research showed that CGA in potatoes is synthesized through HQT and HQT suppression altered phenotype and redirected phenylpropanoid flux.

Synthesis of chlorogenic acid imprinted chromatographic packing by surface-initiated atom transfer radical polymerization and its application

A novel chromatographic packing of chlorogenic acid(CGA) molecularly imprinted polymer(MIP) based on the 5.0 μm silica was prepared by surface initiated atom transfer radical polymerization(SI-ATRP) with 4-vinylpyridine(4-VP) as functional monomer, ethyl glycol dimethacrylate(EDMA) as cross-linker in the mixture of methanol and water(7:3, volume ratio) under mild reaction conditions. The characteristics of CGA MIP were investigated by elemental analysis, thermogravimetric analysis(TGA), Fourier transform infrared spectrometry(FTIR) and atomic force microscopy(AFM). The effects of some chromatographic conditions such as mobile phase composition and temperature on the retention time were investigated. The adsorption capacity of the stationary phase for compounds was determined by frontal chromatographic technique. The results show that Freundlich isotherm fits the experimental adsorption isotherm data better than Langmuir model does. The relatively high heterogeneity index values regressed with the Freundlich isotherm suggest the formation of fairly homogeneous MIP. Thermodynamic data(ΔΔH and ΔΔS) obtained by van’t Hoff plots reveal an entropy-controlled separation. The CGA MIP column was shown to be successful for the separation and purification of chlorogenic acid from the extract of Honeysuckle.