Production Of Chlorogenic Acid Research Articles

Decoding eggplant fruit: Multi-omics profiling of caffeoyl-CoA-3-OMT expression

Eggplant (Solanum melongena L.) is a rich source of health-promoting phenolic acids, mainly chlorogenic acid (CGA), and trace elements. CGA has been shown to have antioxidant, anti-inflammatory, neuroprotective, cardioprotective, anticancer, and antidiabetic properties. The natural genome of eggplants contains essential genes for beneficial traits like drought tolerance, disease resistance, and high concentrations of medicinal substances. Transcriptomic analysis of eggplant overexpressing the hydroxycinnamoyl CoA quinate hydroxycinnamoyl transferase (HQT) gene revealed upregulation of 2165 genes, including three similar to HQT and cinnamate-4-hydroxylase (C4H). Sequence similarity analysis showed homology to caffeoyl-CoA O-methyltransferases, key enzymes in the phenylpropanoid pathway leading to CGA biosynthesis. Gene ontology and KEGG pathway analysis identified 5 highly upregulated glycosyltransferase family 43 genes (650.3-fold change). Glycosyltransferases like UDP-glucose:cinnamate glucosyltransferase and cinnamate-glucose 4′-O-glucosyltransferase are crucial for CGA production in eggplant. qRT-PCR validated the potential upregulation of HQT and C4H in transgenic eggplants. Comparative analysis revealed eggplant has the highest CGA content (5–8.1 g/kg dry weight) among vegetables, with foliar CGA acting as a natural insecticide. CGA offers antioxidant, hypoglycemic, antiviral, and hepatoprotective benefits to humans. This study highlights the role of HQT and C4H in elevating the medicinal properties of eggplant through CGA biosynthesis pathway engineering.

Root endophytic Phialocephala fortinii and Talaromyces verruculosus enhance growth and affect heavy metal tolerance of Miscanthus sinensis Andersson growing naturally at a mine site

ABSTRACT Miscanthus sinensis Andersson naturally grows at mining sites and accumulates heavy metals, suggesting heavy metal tolerance. This study aimed to elucidate the heavy metal tolerance mechanisms of M. sinensis, which naturally grew at a mine site, considering root endophytes. Elemental analysis by inductively coupled plasma-optical emission spectroscopy showed that M. sinensis highly accumulated zinc and lead in adventitious roots and dead root bark, exhibiting tolerance via these elemental removals in dead root bark. Phenolic analysis by high-performance liquid chromatography revealed chlorogenic acid production in adventitious roots. Phialocephala fortinii, which showed the highest appearance, and Talaromyces verruculosus, which produced gluconic acid as a zinc detoxicant, were isolated from adventitious roots of M. sinensis. In inoculation tests, P. fortinii and T. verruculosus significantly enhanced seedling growth by increasing potassium uptake and possibly indole-3-acetic acid production. Our results provide fundamental knowledge for using M. sinensis and root endophytes for revegetation at mining sites.

The Effect of Combined Elicitation with Light and Temperature on the Chlorogenic Acid Content, Total Phenolic Content and Antioxidant Activity of Berula erecta in Tissue Culture.

Chlorogenic acid is one of the most prominent bioactive phenolic acids with great pharmacological, cosmetic and nutritional value. The potential of Berula erecta in tissue culture was investigated for the production of chlorogenic acid and its elicitation combined with light of different wavelengths and low temperature. The content of chlorogenic acid in the samples was determined by HPLC-UV, while the content of total phenolic compounds and the antioxidant activity of their ethanol extracts were evaluated spectrophotometrically. The highest fresh and dry biomasses were obtained in plants grown at 23 °C. This is the first study in which chlorogenic acid has been identified and quantified in Berula erecta. The highest chlorogenic acid content was 4.049 mg/g DW. It was determined in a culture grown for 28 days after the beginning of the experiment at 12 °C and under blue light. The latter also contained the highest content of total phenolic compounds, and its extracts showed the highest antioxidant activity. Berula erecta could, potentially, be suitable for the in vitro production of chlorogenic acid, although many other studies should be conducted before implementation on an industrial scale.

A Single Latent Plant Growth-Promoting Endophyte BH46 Enhances Houttuynia cordata Thunb. Yield and Quality.

Plant growth-promoting endophytes (PGPE) can effectively regulate plant growth and metabolism. The regulation is modulated by metabolic signals, and the resulting metabolites can have considerable effects on the plant yield and quality. Here, tissue culture Houttuynia cordata Thunb., was inoculated with Rhizobium sp. (BH46) to determine the effect of BH46 on H. cordata growth and metabolism, and elucidate associated regulatory mechanisms. The results revealed that BH46 metabolized indole-3-acetic acid and induced 1-aminocyclopropane-1-carboxylate deaminase to decrease ethylene metabolism. Host peroxidase synthesis MPK3/MPK6 genes were significantly downregulated, whereas eight genes associated with auxins, cytokinins, abscisic acid, jasmonic acid, and antioxidant enzymes were significantly upregulated. Eight genes associated with flavonoid biosynthesis were significantly upregulated, with the CPY75B1 gene regulating the production of rutin and quercitrin and the HCT gene directly regulating the production of chlorogenic acid. Therefore, BH46 influences metabolic signals in H. cordata to modulate its growth and metabolism, in turn, enhancing yield and quality of H. cordata.

Mining and functional characterization of NADPH-cytochrome P450 reductases of the DNJ biosynthetic pathway in mulberry leaves

Background1-Deoxynojirimycin (DNJ), the main active ingredient in mulberry leaves, with wide applications in the medicine and food industries due to its significant functions in lowering blood sugar, and lipids, and combating viral infections. Cytochrome P450 is a key enzyme for DNJ biosynthesis, its activity depends on the electron supply of NADPH-cytochrome P450 reductases (CPRs). However, the gene for MaCPRs in mulberry leaves remains unknown.ResultsIn this study, we successfully cloned and functionally characterized two key genes, MaCPR1 and MaCPR2, based on the transcriptional profile of mulberry leaves. The MaCPR1 gene comprised 2064 bp, with its open reading frame (ORF) encoding 687 amino acids. The MaCPR2 gene comprised 2148 bp, and its ORF encoding 715 amino acids. The phylogenetic tree indicates that MaCPR1 and MaCPR2 belong to Class I and Class II, respectively. In vitro, we found that the recombinant enzymes MaCPR2 protein could reduce cytochrome c and ferricyanide using NADPH as an electron donor, while MaCPR1 did not. In yeast, heterologous co-expression indicates that MaCPR2 delivers electrons to MaC3'H hydroxylase, a key enzyme catalyzing the production of chlorogenic acid from 3-O-p-coumaroylquinic acid.ConclusionsThese findings highlight the orchestration of hydroxylation process mediated by MaCPR2 during the biosynthesis of secondary metabolite biosynthesis in mulberry leaves. These results provided a foundational understanding for fully elucidating the DNJ biosynthetic pathway within mulberry leaves.

Partial coding sequence identification, gene expression analysis, and content of anticancer phenolic compounds in Sonchus arvensis L. under drought stress conditions

Field sowthistle, a plant in the Asteraceae family, contains high levels of chlorogenic and chicoric acids, which have anticancer properties. In the current investigation, the expression of genes responsible for chlorogenic acid biosynthesis was studied in S. arvensis in drought stress conditions. The following genes were investigated: phenylalanine ammonia-lyase (PAL), cinnamate 4 hydroxylase (C4H), 4-hydroxycinnamoyl CoA ligase (4CL), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT), hydroxycinnamoyl CoA quinate hydroxycinnamoyl transferase (HQT) and p-coumaroyl ester 3'-hydroxylase (C3'H). As part of a research study, the levels of several anticancer compounds and their correlation with the transcriptional activity of the surveyed genes were investigated under drought stress conditions. The study also involved the identification and submission of the partial coding sequences of the C4H, HCT, C3'H, and actin genes in S. arvensis to the GenBank database. The plants’ middle leaves exposed to 50% field capacity (FC) exhibited the highest expression of all genes. The highest amount of chicoric, chlorogenic, and 2,5 dihydroxybenzoic (2,5 DHB) acids was observed in the middle leaves under 50% FC. In contrast, the highest content of gallic acid and catechin was found in the lower leaves at 75% FC. A strong association between HQT expression and chlorogenic acid content proposed the HQT-dependent metabolic pathway as the primary route for its production in S. arvensis. Furthermore, the consistency between the patterns of gene expression and the levels of compounds found in the middle leaves at 50% FC, except for catechin, showed that the production of chlorogenic acid in S. arvensis could potentially be enhanced by stimulating the expression of the genes that are responsible for its biosynthesis.

Arbuscular mycorrhizal fungal community association determines the production of flavonoids and chlorogenic acid in Acer truncatum Bunge

Arbuscular mycorrhizal (AM) fungi can affect plant secondary metabolic pathways, but how the composition and function of AM fungal communities affect the secondary metabolite content of Acer truncatum Bunge remains unknown. Ten typical A. truncatum plantations were used in southern Shaanxi province. The entropy weight method was applied to construct two grades of A. truncatum tree plantations (ordinary and superior). The interaction between AM fungi and trees in the different grades of plantations was compared. The effects of AM fungal community, soil fertility, climatic factors, and plant nutrients on the contents of flavonoids and chlorogenic acid in leaves were studied. There were no differences in the α and β diversity of AM fungal associations among different grades of trees, but the hyphal colonization rate and co-occurrence network characteristics were different. There were no differences in plant height and diameter at breast height (DBH), but higher flavonoid and chlorogenic acid contents were found in the leaves of superior trees compared to ordinary trees, and this difference was attributed to variations in their AM fungal communities. For A. truncatum trees of similar growth (same height and DBH), the abundance of AM fungi was positively correlated with leaf flavonoids and chlorogenic acid contents in superior tree grade stands, but negatively correlated in ordinary tree stands. The positive correlation between leaf flavonoid content and soil fertility in superior tree grade was mediated by the β diversity of AM fungi recruited in this environment. The relative abundance of VTX00069 and VTX00156 was positively correlated with leaf flavonoid and chlorogenic acid contents, which provides direction for the selection of AM fungi that promote flavonoids and chlorogenic acid production in A. truncatum leaves. Overall, the effect of specific AM fungal communities in the rhizosphere soil of A. truncatum on its flavonoids and chlorogenic acid contents is direct and important. This study provides new insights into enhancing the production of secondary metabolites in A. truncatum.

Efficient Production of Chlorogenic Acid in Escherichia coli Via Modular Pathway and Cofactor Engineering.

Chlorogenic acid is a natural phenolic compound widely used in the food and daily chemical industries. Compared to plant extraction, microbial cell factories provide a green and sustainable production method for the production of chlorogenic acid. However, complex metabolic flux distribution and potential byproducts limited the biosynthesis of chlorogenic acid in microorganisms. A de novo biosynthesis pathway for chlorogenic acid was constructed in Escherichia coli via modular engineering. Increasing the shikimate pathway flux greatly promoted chlorogenic acid production, and the influence of pyruvate metabolism on chlorogenic acid synthesis was also explored. The supply of cofactors for the key enzymes quinate/shikimate 5-dehydrogenase (YdiB) and 4-hydroxyphenylacetate 3-monooxygenase (HpaBC) was enhanced by a cofactor regeneration system. Furthermore, mutants of YdiB were verified for chlorogenic acid production in vivo. Chlorogenic acid browning occurred when the buffer pH of the buffer exceeded 6.0, but two-stage pH control achieved a chlorogenic acid titer of 2789.2 mg/L in a 5 L fermenter, the highest reported to date. This study provided a strategy for the efficient production of chlorogenic acid from simple carbon sources.

Cleavage of macromolecule (protein/polysaccharide)-phenolic bond in soybean cell wall through Lactobacillus casei and Lactobacillus helviticus mixed culture solid-state fermentation for chlorogenic acid extraction

Out of the available dietary sources use for chlorogenic acid extraction, soybean is new and cheap that could offer cost-effective chlorogenic acid production under solid-state fermentation (SSF). Consequently, the study aimed at investigating the cleavage of protein/carbohydrate-phenolic bond in heilong48 soybean (HS) variety via solid-state fermentation by Lactobacillus casei and Lactobacillus helviticus mixed culture for chlorogenic acid extraction with high yield and increased biological activity. A simplistic two-pot bioproduct optimization approach was used; first, screening (Plackett-Burman design) of SSF conditions and second, optimizing (Box-Behnken design) the significant SSF conditions obtained from the former design. The liquid-solid (liquid:solid) ratio of 32:68 (0.47), incubation time of 48 h, pH of 5, and temperature of 50 °C were the best-optimized SSF conditions obtained from the chronological combinatory screening and optimization methods. These conditions gave a satisfactory SSF model, and correspondingly acceptable chlorogenic acid yield (9.20 ± 0.17 mg/g), fermentation efficiency (37.26 ± 0.73%), and 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging activity (DPPH) (65.21 ± 2.05 μmol AA eq/g dry sample) results. The L. casei and L. helviticus mixed culture fermented heilong48 soybean (MCFHS) sample (under optimized SSF conditions) had high (p < 0.05) chlorogenic acid yield and 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging activity than the raw heilong48 soybean flour (RHSF) sample. The effectiveness of the optimized solid-state fermentation conditions on the structural modifications of heilong48 soybean sample was confirmed by scanning electron microscopy, atomic force microscopy, and Fourier transform infrared results.

High-resolution genome mapping and functional dissection of chlorogenic acid production in Lonicera maackii.

Amur honeysuckle (Lonicera maackii) is a widely used medicinal plant of the Caprifoliaceae family that produces chlorogenic acid. Research on this plant mainly focuses on its ornamental value and medicinal compounds, but a reference genome sequence and molecular resources for accelerated breeding are currently lacking. Herein, Nanopore sequencing and high-throughput chromosome conformation capture (Hi-C) allowed a chromosome-level genome assembly of Lonicera maackii (2n = 18). A global view of the gene regulatory network involved in the biosynthesis of chlorogenic acid and the dynamics of fruit coloration in L. maackii was established through metabolite profiling and transcriptome analyses. Moreover, we identified the genes encoding hydroxycinnamoyl-CoA quinate transferase (LmHQT) and hydroxycinnamoyl-CoA shikimic/quinate transferase (LmHCT), which localized to the cytosol and nucleus. Heterologous overexpression of these genes in Nicotiana benthamiana leaves resulted in elevated chlorogenic acid contents. Importantly, HPLC analyses revealed that LmHCT and LmHQTs recombinant proteins modulate the accumulation of chlorogenic acid (CGA) using quinic acid and caffeoyl CoA as substrates, highlighting the importance of LmHQT and LmHCT in chlorogenic acid biosynthesis. These results confirmed that LmHQTs and LmHCT catalyze the biosynthesis of CGA in vitro. The genomic data presented in this study will offer a valuable resource for the elucidation of CGA biosynthesis and facilitating selective molecular breeding.

Exploring the metabolic and physiological roles of HQT in S. lycopersicum by gene editing.

The most abundant phenolic compound in Solanaceous plants is chlorogenic acid (CGA), which possesses protective properties such as antimicrobial and antioxidant activities. These properties are particularly relevant when plants are under adverse conditions, such as pathogen attack, excess light, or extreme temperatures that cause oxidative stress. Additionally, CGA has been shown to absorb UV-B light. In tomato and potato, CGA is mainly produced through the HQT pathway mediated by the enzyme hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase. However, the absence of natural or induced mutants of this gene has made it unclear whether other pathways contribute to CGA production and accumulation. To address this question, we used CRISPR technology to generate multiple knock-out mutant lines in the tomato HQT gene. The resulting slhqt plants did not accumulate CGA or other caffeoylquinic acids (CQAs) in various parts of the plant, indicating that CQA biosynthesis depends almost entirely on the HQT pathway in tomato and, likely, other Solanaceous crops. We also found that the lack of CGA in slhqt plants led to higher levels of hydroxycinnamoyl-glucose and flavonoids compared to wild-type plants. Gene expression analysis revealed that this metabolic reorganization was partly due to flux redirection, but also involved modulation of important transcription factor genes that regulate secondary metabolism and sense environmental conditions. Finally, we investigated the physiological role of CGA in tomato and found that it accumulates in the upper epidermis where it acts as a protector against UV-B irradiation.

Regulatory network characterization of anthocyanin metabolites in purple sweetpotato via joint transcriptomics and metabolomics.

Sweet potato is an important staple food crop in the world and contains abundant secondary metabolites in its underground tuberous roots. The large accumulation of several categories of secondary metabolites result in colorful pigmentation of the roots. Anthocyanin, is a typical flavonoid compound present in purple sweet potatoes and it contributes to the antioxidant activity. In this study, we developed joint omics research via by combing the transcriptomic and metabolomic analysis to explore the molecular mechanisms underlying the anthocyanin biosynthesis in purple sweet potato. Four experimental materials with different pigmentation phenotypes, 1143-1 (white root flesh), HS (orange root flesh), Dianziganshu No.88 (DZ88, purple root flesh), and Dianziganshu No.54 (DZ54, dark purple root flesh) were comparably studied. We identified 38 differentially accumulated pigment metabolites and 1214 differentially expressed genes from a total of 418 metabolites and 50893 genes detected. There were 14 kinds of anthocyanin detected in DZ88 and DZ54, with glycosylated cyanidin and peonidin as the major components. The significantly enhanced expression levels of multiple structural genes involved in the central anthocyanin metabolic network, such as chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase/leucocyanidin oxygenase (ANS), and glutathione S-transferase (GST) were manifested to be the primary reason why the purple sweet potatoes had a much higher accumulation of anthocyanin. Moreover, the competition or redistribution of the intermediate substrates (i.e. dihydrokaempferol and dihydroquercetin) between the downstream production of anthocyanin products and the flavonoid derivatization (i.e. quercetin and kaempferol) under the regulation of the flavonol synthesis (FLS) gene, might play a crucial role in the metabolite flux repartitioning, which further led to the discrepant pigmentary performances in the purple and non-purple materials. Furthermore, the substantial production of chlorogenic acid, another prominent high-value antioxidant, in DZ88 and DZ54 seemed to be an interrelated but independent pathway differentiated from the anthocyanin biosynthesis. Collectively, these data from the transcriptomic and metabolomic analysis of four kinds of sweet potatoes provide insight to understand the molecular mechanisms of the coloring mechanism in purple sweet potatoes.

Optimization of Glycerol Extraction of Chlorogenic Acid from Honeysuckle by Response Surface Methodology

Using honeysuckle as raw material, chlorogenic acid (CGA) was extracted with different alcohols. Based on the single-factor experiment design, the relationship between each parameter and the response value was explored by Box–Behnken method to optimize the process conditions. Best extraction results were obtained under the conditions of solid-to-liquid ratio of 1:20, the ultrasonic time of 40 min, the ultrasonic vibrator power of 240 w, and the CGA extraction rate of 2.98%. The experimental data show that the extraction rate of CGA is related to the length of the alcohol carbon chain and the number of hydroxyl groups in the extractant. The results from this work can provide technical basis for the safe and efficient production of CGA from honeysuckle.

Gene Expression and Metabolic Analyses of Nontransgenic and AtPAP1 Transgenic Tobacco Infected with Potato Virus X (PVX).

Potato virus X (PVX), a species of the genus Potexvirus, is a plant pathogenic virus that causes severe symptoms such as mild mosaic, crinkling, necrosis, and mottling on leaves. The objectives of the present study were to investigate the effect of PVX virus infection on the metabolic system in nontransgenic and Arabidopsis thaliana production of anthocyanin pigment 1 (AtPAP1) transgenic tobacco using transcript expression analysis and metabolic profiling. Potato virus X inoculation increased the gene expression of phenylpropanoid and flavonoid biosynthesis and the production of chlorogenic acid, p-coumaric acid, benzoic acid, rutin, quercetin, and kaempferol in nontransgenic tobacco leaves. However, in the AtPAP1 transgenic tobacco leaves, PVX inoculation decreased the expression of AtPAP1 and phenylpropanoid and flavonoid biosynthesis genes, and the production of phenolics and anthocyanin also declined. In contrast, the levels of amino acids and tricarboxylic acid (TCA) cycle intermediates increased after infection in the AtPAP1 transgenic plant leaves. To date, these results have not been reported previously. We suggest that PVX infection decreases AtPAP1 expression, leading to the downregulation of phenylpropanoid and flavonoid biosynthesis in transgenic plants.

Establishment of bioreactor cultures for the production of chlorogenic acid and ferulic acid from adventitious roots by optimization of culture conditions in Angelica acutiloba (Siebold &amp; Zucc.) Kitag

Establishment of bioreactor cultures for the production of chlorogenic acid and ferulic acid from adventitious roots by optimization of culture conditions in Angelica acutiloba (Siebold &amp; Zucc.) Kitag

Metabolic Engineering of Saccharomyces cerevisiae for High-Level Production of Chlorogenic Acid from Glucose.

Chlorogenic acid (CGA), a major dietary phenolic compound, has been increasingly used in the food and pharmaceutical industries because of its ready availability and extensive biological and pharmacological activities. Traditionally, extraction from plants has been the main approach for the commercial production of CGA. This study reports the first efficient microbial production of CGA by engineering the yeast, Saccharomyces cerevisiae, on a simple mineral medium. First, an optimized de novo biosynthetic pathway for CGA was reconstructed in S. cerevisiae from glucose with a CGA titer of 36.6 ± 2.4 mg/L. Then, a multimodule engineering strategy was employed to improve CGA production: (1) unlocking the shikimate pathway and optimizing carbon distribution; (2) optimizing the l-Phe branch and pathway balancing; and (3) increasing the copy number of CGA pathway genes. The combination of these interventions resulted in an about 6.4-fold improvement of CGA titer up to 234.8 ± 11.1 mg/L in shake flask cultures. CGA titers of 806.8 ± 1.7 mg/L were achieved in a 1 L fed-batch fermenter. This study opens a route to effectively produce CGA from glucose in S. cerevisiae and establishes a platform for the biosynthesis of CGA-derived value-added metabolites.

Polyscias filicifolia (Araliaceae) Hairy Roots with Antigenotoxic and Anti-Photogenotoxic Activity.

Hairy root cultures are considered as a valuable source of bioactive phytoconstituents with expanding applicability for their production. In the present study, hairy root cultures of Polyscias filicifolia (Araliaceae), a traditional Southeast Asian medicinal plant, were established. The transformation with Agrobacterium rhizogenes ATCC 15834 allowed to obtain 15 root lines. The K-1 line, demonstrating the highest growth capabilities, was subjected to further investigations. To enhance the biosynthetic potential of hairy roots, methyl jasmonate elicitation approach was applied (MeJA; at different doses and exposure time), with subsequent transfer of elicited roots to control medium. This strategy resulted in chlorogenic acid production up to 1.59 mg/g dry weight. HPLC-PDA-ESI-MS analysis demonstrated variation in extracts composition and allowed to identify different caffeic and ferulic acid derivatives. Next, cytotoxic, antigenotoxic, and anti-photogenotoxic properties of hairy roots extracts were determined. None of the tested extracts were cytotoxic. In addition, they demonstrated significant antigenotoxic activity with the highest protective potential; up to 52% and 49% of inhibition of induction ratio (IR) induced by the 2-aminoanthracene was revealed for extracts derived from hairy roots elicited for 3 days with 50 µM MeJA and roots elicited for 7 days with 100 µM MeJA and then transferred for 30 days to control medium, respectively. These same extracts exhibited the highest anti-photogenotoxic potential, up to 36% of inhibition of chloropromazine-induced genotoxicity.

Cell-free Biosynthesis of Chlorogenic Acid Using a Mixture of Chassis Cell Extracts and Purified Spy-Cyclized Enzymes.

A novel cell-free biosynthesis system based on a mixture of chassis cell extracts and purified Spy-cyclized enzymes (CFBS-mixture) was developed. As a demonstration, the CFBS-mixture was applied to chlorogenic acid (CGA) biosynthesis. The mix-and-match and Plackett-Burman experiments demonstrated that Lonicera japonica hydroxycinnamate-CoA quinate transferase and p-hydroxyphenylacetate 3-hydroxylase were the key enzymes for the production of CGA. After optimization of the concentrations of the biosynthetic enzymes in the CFBS-mixture reaction using the Plackett-Burman experimental design and the path of the steepest ascent, 711.26 ± 15.63 mg/L CGA was produced after 16 h, which is 71.1-fold the yield obtained using the conventional crude extract-based CFBS and 9.1-fold the reported yield obtained using the living cells. Based on the CFBS-mixture results, the production of CGA was further enhanced in engineered Escherichia coli. The CFBS-mixture strategy is highly effective and will be useful for high-level CFBS of natural products.

Salicylic acid inhibits the postharvest decay of goji berry (Lycium barbarum L.) by modulating the antioxidant system and phenylpropanoid metabolites

Goji berry is a perishable fruit after harvest. To explore the biochemical and molecular mechanisms by which salicylic acid (SA) suppresses rot in goji berry, the fruit was treated with SA and stored for 30 d. Compared with the control, the SA-treated fruit showed smaller lesion areas during storage at 20 °C. During storage at 0 °C, the O2∙¯ production and H2O2 levels were lower in SA-treated fruit, although the H2O2 content was higher at the beginning of storage. SA induced the activities and gene expressions of the key proteins (SOD, CAT, APX and POD) associated with redox homeostasis. Simultaneously, the activities and gene expressions of PAL, C4H, 4CL, CHS, CHI and CAD in phenylpropanoid metabolism was activated. Moreover, the production of chlorogenic acid, ferulic acid, p-coumaric acid, sinapic acid, protocatechuic acid, naringenin, and rutin were stimulated to maintain the higher levels by the most effective SA treatment (2 mmol L−1). The results indicate that SA inhibits rot of goji fruit during storage by mediating the antioxidant system and phenylpropanoid metabolism.

Nanosilicon enhances maize resistance against oriental armyworm (Mythimna separata) by activating the biosynthesis of chemical defenses.

Silicon, in its nanoscale form, has shown plant-promoting and insecticidal properties. To date, however, we lack mechanistic evidence for how nanoscale silicon influences the regulation of plant chemical defenses against herbivore attacks. To address this gap, we compared the effect of Si nanodots (NDs) and sodium silicate, a conventional silicate fertilizer, on maize (Zea mays L.) chemical defenses against the oriental armyworm (Mythimna separata, Walker) caterpillars. We found that Si NDs and sodium silicate additions, at the dose of 50mg/L, significantly inhibited the growth of caterpillars by 53.5% and 34.2%, respectively. This increased plant resistance was associated with a 44.2% increase in the production of chlorogenic acid, as well as the expression of PAL, C4H, 4CL, C3H and HCT, core genes involved in the biosynthesis of chlorogenic acid, by 1.7, 2.4, 1.9, 1.8 and 4.5 folds, respectively. Particularly, in the presence of M. separata, physiological changes in maize plants treated with 50mg/L Si NDs, including changes in shoot biomass, leaf nutrients (e.g., K, P, Si), and chemical defense compounds (e.g., chlorogenic acid, total phenolics), were higher than those of plants added with equivalent concentrations of conventional silicate fertilizer. Taken together, our findings indicate that Si, in nanoscale form, could replace synthetic pesticides, and be implemented for a more effective and ecologically-sound management of insect pests in maize crop farming.