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Abstract Background Amomum tsao-ko is an important aromatic crop used in medicines and food. It can be categorized into three main types based on the fruit shape: long (L), oval (O), and round (R). However, limited information is available on the volatile substances present in differently shaped A. tsao-ko fruits. This study investigated the characteristics and biosynthesis of volatile organic compounds (VOCs) in fresh and dried A. tsao-ko fruits of different shapes using widely targeted volatilomics and transcriptome analyses. Results In total, 978 VOCs, primarily terpenoids, esters, and heterocyclic compounds, were detected. The number of differentially accumulated volatile organic compounds (DAVOCs) in dried fruits of various shapes was significantly higher than that in fresh fruits, with terpenoids, esters, and heterocyclic compounds accounting for approximately 50% of the total DAVOCs. Notably, α-phellandrene, identified as a shared differential accumulated terpenoid across various fruit shapes, was detected in both fresh and dried fruits. Through transcriptome analysis, 40 candidate genes implicated in the terpenoid biosynthesis pathway were screened. An integrated analysis of the metabolome and transcriptome revealed that the structural genes HMGR-2, TPS7, TPS5-10, TPS21-3, TPS21-5, TPS21-6, TPS21-7, and TPS21-9, along with 81 transcription factors (including 17 NACs, 16 MYBs, 16 AP2/ERFs, 13 WRKYs, 13 bHLHs, and 6 bZIPs), co-regulate the biosynthesis of volatile terpenoids. Conclusions This study expands our understanding of the volatile metabolism profile of A. tsao-ko and provides a solid foundation for future investigations of the mechanisms governing fruit quality.

Context: Interleukin-17 (IL-17) is a pro-inflammatory cytokine that plays a crucial role in immunity and inflammation. Aims: To evaluate the potential therapeutic effects of two compounds, magnosalicine and neotatarine, derived from the ethanolic extract of Acorus calamus L. rhizome, against IL-17. Methods: Utilizing LC/MS analysis combining molecular docking simulations, drug likeness, ADME and toxicity analysis. This study explored the molecular interactions and binding affinities of identified compounds from A. calamus with crucial residues of IL-17, including Tyr62, Pro63, Ile66, Gln94, Ile96, Leu97, and Leu99. Results: The results revealed that magnosalicine and neotatarine exhibited remarkable binding affinities of -10.16 kcal/mol and -9.53 kcal/mol, respectively, indicating their strong interactions with IL-17. Moreover, both compounds displayed superior binding energies compared to other extract constituents. Interestingly, this study highlighted that all terpenoid compounds from the A. calamus rhizome extract were capable of interacting with these key residues of IL-17, resembling the interactions observed with the natural ligand (RMK) and methotrexate. Meanwhile, the analysis results revealed a safer ADME and toxicity profile for neotatarine compared to magnolalicine. Conclusions: This research unveils the promising potential of neotatarine as candidates for further exploration in therapeutic interventions targeting IL-17-related pathways. These findings shed light on the molecular insights of Acorus calamus L. compounds, providing valuable information for developing novel treatments for IL-17-associated disorders.

Terpenoids are one of the largest class of natural products with diverse structures and activities. This enormous diversity is embedded in enzymes called terpene synthases (TSs), which generate diverse terpene skeletons via sophisticated cyclization cascades. In addition to the many highly selective TSs, there are many promiscuous TSs that accept multiple prenyl substrates, or even noncanonical ones, with 6, 7, 8, 11, and 16 carbon atoms, synthesized via chemical approaches, C-methyltransferases, or engineered lepidopteran mevalonate pathways. The substrate promiscuity of TSs not only expands the structural diversity of terpenes but also highlights their potential for the discovery of novel terpenoids via combinatorial biosynthesis. In this review, we focus on the current knowledge on multisubstrate terpene synthases (MSTSs) and highlight their potential applications.

As an important spice species in Rutaceae, the Sichuan pepper (Zanthoxylum armatum) can provide pungent and numbing taste, as well as aroma in its mature fruit. Here we assembled a chromosome-level genome of green prickly ash which was widely cultivated in a major production area including Chongqing and Sichuan province, China. We generated 712 Gb (∼112 × ) PacBio long reads and 511 Gb (∼82 × ) Hi-C data, and yielded an assembly of 99 pseudochromosomes with total size of 5.32 Gb and contig N50 of 796 kb. The genomic analyses and cytogenetic experiments both indicated that the cultivar ‘Zhuye Huajiao’ was a triploid. We identified a Zanthoxylum-specific whole genome duplication event emerging about 24.8 million years ago (Mya). We also detected a transposition burst event (0.3–0.4 Mya) responsible for the large genome size of Z. armatum. Metabolomic analysis of the Zanthoxylum fruits during development stages revealed profiles of 39 volatile aroma compounds and 528 secondary metabolites, from which six types of sanshools were identified. Based on metabolomic and transcriptomic network analysis, we screened candidate genes encoding long chain acyl-CoA synthetase, fatty acid desaturase, branched-chain amino acid aminotransferase involved in sanshool biosynthesis and three genes encoding terpene synthase during fruit development. The multi-omics data provide insights into the evolution of Zanthoxylum and molecular basis of numbing and aroma flavor of Sichuan pepper.

IntroductionCamphora longepaniculata, a crucial commercial crop and a fundamental component of traditional Chinese medicine, is renowned for its abundant production of volatile terpenoids. However, the lack of available genomic information has hindered pertinent research efforts in the past.MethodsTo bridge this gap, the present study aimed to use PacBio HiFi, short-read, and highthroughput chromosome conformation capture sequencing to construct a chromosome-level assembly of the C. longepaniculata genome. Results and discussionWith twelve chromosomes accounting for 99.82% (766.69 Mb) of the final genome assembly, which covered 768.10 Mb, it was very complete. Remarkably, the assembly’s contig and scaffold N50 values are exceptional as well—41.12 and 63.78 Mb, respectively—highlighting its excellent quality and intact structure. Furthermore, a total of 39,173 protein-coding genes were predicted, with 38,766 (98.96%) of them being functionally annotated. The completeness of the genome was confirmed by the Benchmarking Universal Single-Copy Ortholog evaluation, which revealed 99.01% of highly conserved plant genes. As the first comprehensive assembly of the C. longepaniculata genome, it provides a crucial starting point for deciphering the complex pathways involved in terpenoid production. Furthermore, this excellent genome serves as a vital resource for upcoming research on the breeding and genetics of C. longepaniculata.

Ferula L. plants are of great importance in a number of fields such as agriculture, veterinary, medicine and food industry. Medicinal properties of Ferula L. species are explained by the variety of secondary metabolites they produce, in which the activity of root rhizobacteria occupies a special place. This article presents the research-based analysis of some biological properties, agrotechnology, secondary metabolites (ferulin A,D; diversolid A,D,F,G; kuhistanicaol A,B,C,D,G; kuhistanol (A-H); fucanefuromarin (A-G); fucanemarin A,B; fesumtuorin (A-H); ferulagol A,B; pallidon (A-F); sinkiangenorin D and others) of the species of genus Ferula L., the influence of the microbial communities (new species belonging to the genera Porphyrobacter, Paracoccus and Amycolatopsis, as well as Actinobacteria, Acidobacteria, Proteobacteria, Gemmatimonadet and Bacteroides) living in the roots of Ferula L. on the synthesis of some secondary metabolites of the plant, based on scientific sources.

Abstract Plant secondary metabolites can protect organisms against oxidative stress caused by adverse environmental conditions. Cinnamomum camphora (L.) J. Presl contains plentiful terpenoids and is subdivided into 5 chemotypes. To develop natural antioxidants using the plant terpenoids, the terpenoid composition and antioxidant abilities of methanol, ethyl acetate, n-hexane and petroleum ether extracts from linalool and eucalyptol chemotypes were investigated, and the solvent effects on in vitro antioxidant activity of 8 main terpenoids were analyzed. Meanwhile, the in vivo effects of two strong antioxidant terpenoids were evaluated. For the two chemotypes, the 4 solvents exhibited the same extracting effects on the terpenoid types, but methanol extracts contained the highest content of terpenoids, which should contribute to their strongest scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals. In linalool and eucalyptol chemotype extracts, linalool, eucalyptol, ocimene, α-pinene, D-limonene, terpinene, β-pinene and longifolene were the 8 main terpenoids. Among the 4 solvents, the 8 terpenoids showed the strongest free radical-scavenging activity with methanol as the reaction medium, which might result from strong-polarity methanol easily activating C = C unsaturated bonds in these compounds. This might also contribute to the strongest scavenging activity of methanol extracts against free radicals. Among these main components, ocimene and longifolene separately showed the strongest activity in scavenging DPPH and ABTS free radical. In in vivo assay, the two compounds significantly lowered the reactive oxygen species (ROS) levels in Chlamydomonas reinhardtii, a single-celled model organism, under H2O2 stress, and the lowering effects gradually enhanced with increasing the compound concentration, resulting in the corresponding promoting effect on the algal growth. At the same concentration, longifolene showed the strongest effect on protecting the algal cells against oxidative stress. Therefore, methanol was suitable for extracting terpenoids in natural antioxidant development, and ocimene and longifolene were two strong antioxidant terpenoids without cell toxicity, with the latter having stronger in vivo antioxidant activity. Graphical Abstract

Abstract Background Cannabis sativa, a dioecious plant that has been cultivated worldwide for thousands of years, is known for its secondary metabolites, especially cannabinoids, which possess several medicinal effects. In this study, we investigated the autopolyploidization effects on the biosynthesis and accumulation of these metabolites, transcriptomic and metabolomic analyses were performed to explore the gene expression and metabolic variations in industrial hemp autotetraploids and their diploid progenitors. Results Through these analyses, we obtained 1,663 differentially expressed metabolites and 1,103 differentially expressed genes. Integrative analysis revealed that phenylpropanoid and terpenoid biosynthesis were regulated by polyploidization. No substantial differences were found in the cannabidiol or tetrahydrocannabinol content between tetraploids and diploids. Following polyploidization, some transcription factors, including nine bHLH and eight MYB transcription factors, affected the metabolic biosynthesis as regulators. Additionally, several pivotal catalytic genes, such as flavonol synthase/flavanone 3-hydroxylase, related to the phenylpropanoid metabolic pathway, were identified as being modulated by polyploidization. Conclusions This study enhances the overall understanding of the impact of autopolyploidization in C. sativa and the findings may encourage the application of polyploid breeding for increasing the content of important secondary metabolites in industrial hemp.

Basil (Ocimum basilicum L.) is a widely used culinary herb. In this study, ethanol, dichloromethane, and sunflower oil were used separately as solvents with distinct polarities for the extraction of basil aerial parts to simulate the different polarity conditions in domestic food processing. The oil extract (OE) was re-extracted with acetonitrile, and the chemical composition, antioxidant potential, and antimicrobial activities of the ethanol (EE), dichloromethane (DCME), and acetonitrile (ACNE) extracts were determined. A total of 109 compounds were tentatively identified in EE, DCME, and ACNE by HPLC–DAD/ESI-ToF-MS. Fatty acids were present in all extracts. Phenolic acids and flavonoids dominated in EE. DCME was characterised by triterpenoid acids, while diterpenoids were mainly found in ACNE. The extracts were analysed for their antioxidant capacity using the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay. EE and DCME showed significant radical scavenging potential. Antimicrobial activity was explored in eight bacterial, two yeast, and one fungal species. All extracts exhibited high antifungal activity, comparable to or better than that of the commercial drug nistatin. Antibacterial activities were notable for EE and ACNE, while DCME showed no activity against bacteria in the applied concentration ranges. The different polarities of the solvents led to distinctive phytochemical compositions and bioactivities in the extracts.

Nanoscale three-dimensional (3D) network morphologies, such as those represented by gyroids accessed by the self-assembly of block copolymers, are highly attractive platforms for the fabrication of nanostructured materials. However, it remains challenging to access such morphologies, especially in the sub-10 nm domain spacing region. Herein, we systematically investigated the microphase separation behaviors of biobased sugar–terpenoid hybrid block co-oligomers (BCOs) with different molecular parameters, including their volume fractions, the linker structures between the blocks, and the stereochemistries of the terpenoid blocks. The BCOs were synthesized using the azido–alkyne click reaction of propargyl-β-d-glucopyranoside with azido-functionalized farnesol, phytol, DL-α-tocopherol, and d-α-tocopherol, to ensure a well-defined molecular structure without any molecular weight distribution. Through X-ray scattering screening, we found that tocopherols, when combined with a glucose unit, yield BCOs capable of forming gyroid and hexagonally perforated lamellar (HPL) 3D network morphologies with ultrasmall domain spacings of ∼10 nm. Remarkably, HPL, which is known as a metastable phase in block copolymers, was obtained in both the bulk and film states. The nature of the linker structure and the stereochemistry of the tocopherol hydrocarbon chain were found to influence the resulting morphology and the long-range order of the nanostructures. We expect this study to contribute to the molecular design of precise block copolymers and the construction of intricate nanostructural templates with ultrasmall feature sizes.

Context: Curcuma xanthorrhiza, a native to Indonesia, has been recognized as a potential medicinal plant. Xanthorrhizol (XTZ), curcumin (Cur), bisdemethoxycurcumin (BDMC), and demethoxycurcumin (DMC) are the main major metabolites with different bioactivities. Thus, the selective extraction of these markers is required. Aims: To characterize and use natural deep eutectic solvents (NADES) for the extraction of XTZ and curcuminoids (CURS) from C. xanthorrhiza. Methods: Four NADES, comprising of glucose:lactic acid (Glu:LA, 1:3), choline chloride: malic acid (ChCl:MA, 1:1), choline chloride: glycerol (ChCl:Gly, 1:1), and choline chloride: sucrose (ChCl:Suc, 3:1) were synthesized and subjected to FTIR. The physicochemical properties of NADES were characterized, including polarity, viscosity, and density. Their antioxidant profiles were evaluated. The toxicity of NADES was assessed towards Escherichia coli by real-time growth monitoring. Tested NADES were employed to extract marker compounds from the rhizome of C. xanthorrhiza. A validated high-performance liquid chromatography (HPLC) method was applied to simultaneously quantify XTZ and CURS in NADES extracts. Results: FTIR spectra confirmed the formation of hydrogen bonds between components. Using the DPPH and FRAP methods, all NADES and NADES extracts displayed antioxidative activity, suggesting the contribution of NADES to the activity. Of the tested NADES, Glu:LA (1:3) obtained the highest content of XTZ over Cur, BDMC, and DMC, comparable to that of ethanol maceration. All tested NADES were harmless to E. coli, except for ChCl:MA (1:1). However, NADES extracts of Glu:LA (1:3) and ChCl:MA (1:1) suppressed the growth of the bacterium. Scanning electron micrographs demonstrated the destruction of the surface structure of the rhizome powder by NADES-ultrasonication. Conclusions: The study recommended the use of Glu:LA (1:3) to selectively extract XTZ to promote environmentally friendly and sustainable extraction.

Sorbitol is an important source of abiotic stress that is used to increase osmolality in cell cultures. It increases the antioxidant enzymes of defense catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in the stress state of cells. Sorbitol plays an important role in stimulating these enzymes in cells and increasing phenylalanine ammonium lyase (PAL) activity. The aim of this study was to apply increasing doses of sorbitol elicitor to cell suspension cultures to determine the changes in cell number, viability, dry weight, and camphor content. In vitro plantlets were obtained from plant seeds and stem segments of these plants were used as explant source. Cell cultures were established after callus formation. Then, 0 (control), 5, 25, and 50 g L-1 sorbitol was dissolved in distilled water and cultured. Samples were taken three times in total, starting from day 1 to day 3. The content of camphor was detected by gas chromatography-mass spectrometry (GC-MS). Cell number, viability,dry weight, and camphor content increased significantly with increasing doses of sorbitol compared to sampling times. Compared to the initial culture, the amount of camphor increased by 40% at the 5 g L-1 dose, 82% at the 25 g L-1 dose, and 154% at the 50 g L-1 dose. In A. gypsicola cell cultures, increasing doses of sorbitol have clearly demonstrated the secondary metabolite accumulation and its positive effect on cell growth.

Cadmium (Cd) contamination poses a substantial threat the environment, necessitating effective remediation strategies. Phytoremediation emerges as a cost-efficient and eco-friendly approach for reducing Cd levels in the soil. In this study, the suitability of A. venetum for ameliorating Cd-contaminated soils was evaluated. Mild Cd stress promoted seedling and root growth, with the root being identified as the primary tissue for Cd accumulation. The Cd content of roots ranged from 0.35 to 0.55 mg/g under treatment with 10–50 µM CdCl2·2.5 H2O, and the bioaccumulation factor ranged from 28.78 to 84.43. Transcriptome sequencing revealed 20,292 unigenes, and 7507 nonredundant differentially expressed genes (DEGs) were identified across five comparison groups. DEGs belonging to the “MAPK signaling pathway-plant,” “monoterpenoid biosynthesis,” and “flavonoid biosynthesis pathway” exhibited higher expression levels in roots compared to stems and leaves. In addition, cytokinin-related DEGs, ROS scavenger genes, such as P450, glutathione-S-transferase (GST), and superoxide dismutase (SOD), and the cell wall biosynthesis-related genes, CSLG and D-GRL, were also upregulated in the root tissue, suggesting that Cd promotes root development. Conversely, certain ABC transporter genes, (e.g, NRAMP5), and some vacuolar iron transporters, predominantly expressed in the roots, displayed a strong correlation with Cd content, revealing the mechanism underlying the compartmentalized storage of Cd in the roots. KEGG enrichment analysis of DEGs showed that the pathways associated with the biosynthesis of flavonoids, lignin, and some terpenoids were significantly enriched in the roots under Cd stress, underscoring the pivotal role of these pathways in Cd detoxification. Our study suggests A. venetum as a potential Cd-contaminated phytoremediation plant and provides insights into the molecular-level mechanisms of root development promotion and accumulation mechanism in response to Cd stress.

Abstract Enzyme spatial organization is an evolved mechanism for facilitating multi‐step biocatalysis and can play an important role in the regulation of promiscuous enzymes. The latter function suggests that artificial spatial organization can be an untapped avenue for controlling the specificity of bioengineered metabolic pathways. A promiscuous terpene synthase (nerolidol synthase) is co‐localized and spatially organized with the preceding enzyme (farnesyl diphosphate synthase) in a heterologous production pathway, via translational protein fusion and/or co‐encapsulation in a self‐assembling protein cage. Spatial organization enhances nerolidol production by ≈11‐ to ≈62‐fold relative to unorganized enzymes. More interestingly, striking differences in the ratio of end products (nerolidol and linalool) are observed with each spatial organization approach. This demonstrates that artificial spatial organization approaches can be harnessed to modulate the product profiles of promiscuous enzymes in engineered pathways in vivo. This extends the application of spatial organization beyond situations where multiple enzymes compete for a single substrate to cases where there is competition among multiple substrates for a single enzyme.

Objective: The objective of this study is to find out the role of terpenoid compounds as potential inhibitors against certain protein targets of dengue virus. Methods: The 2-dimensional structures of terpenoid compounds were retrieved from the PubChem database. They were analysed for their interactions with target proteins of dengue virus such as envelope (1OKE), NS5 (1R6A-RPV & SHA) and NS2B/NS3 (4M9K) by docking studies followed by molecular dynamics (MD) simulations using Schrödinger software, version 10.7. Results: Out of 513 terpenoid compounds studied, malacitanolide showed the highest interaction energy values of −7.072 kcal/mol (hydrogen bond (HB) interactions with Thr280, Gln200, Gln271, Gln49 and Ala50), −5.295 kcal/mol (HB interactions with Ser150, Lys29, Ser214) and −4.030 kcal/mol (HB interactions with Glu1169 and Asn1119) against 1OKE, 1R6A-RPV and 4M9K targets, respectively. Paclitaxel had shown the interaction energy value of −9.334 kcal/mol (HB with Lys61, Asp146, Trp87, Gly148, and Arg84) with 1R6A-SAH. MD simulation studies revealed that the best interacting compound malacitanolide maintained a stable complex with 1OKE of dengue virus. Malacitanolide, reissantin E, and paclitaxel exhibited very good interactions with all three-protein targets of dengue virus and had also shown significant stability. Conclusions: In the present study, it is concluded that the terpenoid compounds malacitanolide, reissantin E, and paclitaxel could act as potential inhibitors against all three target proteins of dengue virus.

Medicinal plants are rich in secondary metabolites with beneficial pharmacological effects. The production of plant secondary metabolites is subjected to the influences by environmental factors including the plant-associated microbiome, which is crucial to the host’s fitness and survival. As a result, research interests are increasing in exploiting microbial capacities for enhancing plant production of pharmacological metabolites. A growing body of recent research provides accumulating evidence in support of developing microbe-based tools for achieving this objective. This mini review presents brief summaries of recent studies on medicinal plants that demonstrate microbe-augmented production of pharmacological terpenoids, polyphenols, and alkaloids, followed by discussions on some key questions beyond the promising observations. Explicit molecular insights into the underlying mechanisms will enhance microbial applications for metabolic fortification in medicinal plants.

AbstractAntrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

Little is known concerning terpenoids produced by members of the fungal order Ophiostomales, with the member Harringtonia lauricola having the unique lifestyle of being a beetle symbiont but potentially devastating tree pathogen. Nine known terpenoids, including six labdane diterpenoids (1–6) and three hopane triterpenes (7–9), were isolated from H. lauricola ethyl acetate (EtOAc) extracts for the first time. All compounds were tested for various in vitro bioactivities. Six compounds, 2, 4, 5, 6, 7, and 9, are described functionally. Compounds 2, 4, 5, and 9 expressed potent antiproliferative activity against the MCF-7, HepG2 and A549 cancer cell lines, with half-maximal inhibitory concentrations (IC50s) ~12.54–26.06 μM. Antimicrobial activity bioassays revealed that compounds 4, 5, and 9 exhibited substantial effects against Gram-negative bacteria (Escherichia coli and Ralstonia solanacearum) with minimum inhibitory concentration (MIC) values between 3.13 and 12.50 μg/mL. Little activity was seen towards Gram-positive bacteria for any of the compounds, whereas compounds 2, 4, 7, and 9 expressed antifungal activities (Fusarium oxysporum) with MIC values ranging from 6.25 to 25.00 μg/mL. Compounds 4, 5, and 9 also displayed free radical scavenging abilities towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide (O2−), with IC50 values of compounds 2, 4, and 6 ~3.45–14.04 μg/mL and 22.87–53.31 μg/mL towards DPPH and O2−, respectively. These data provide an insight into the biopharmaceutical potential of terpenoids from this group of fungal insect symbionts and plant pathogens.

Long non-coding RNAs (lncRNAs) have emerged as curial regulators of diverse biological processes in plants. Jasmine (Jasminum sambac) is a world-renowned ornamental plant for its attractive and exceptional flower fragrance. However, to date, no systematic screening of lncRNAs and their regulatory roles in the production of the floral fragrance of jasmine flowers has been reported. In this study, we identified a total of 31,079 novel lncRNAs based on an analysis of strand-specific RNA-Seq data from J. sambac flowers at different stages. The lncRNAs identified in jasmine flowers exhibited distinct characteristics compared with protein-coding genes (PCGs), including lower expression levels, shorter transcript lengths, and fewer exons. Certain jasmine lncRNAs possess detectable sequence conservation with other species. Expression analysis identified 2752 differentially expressed lncRNAs (DE_lncRNAs) and 8002 DE_PCGs in flowers at the full-blooming stage. DE_lncRNAs could potentially cis- and trans-regulate PCGs, among which DE_lincRNAs and their targets showed significant opposite expression patterns. The flowers at the full-blooming stage are specifically enriched with abundant phenylpropanoids and terpenoids potentially contributed by DE_lncRNA cis-regulated PCGs. Notably, we found that many cis-regulated DE_lncRNAs may be involved in terpenoid and phenylpropanoid/benzenoid biosynthesis pathways, which potentially contribute to the production of jasmine floral scents. Our study reports numerous jasmine lncRNAs and identifies floral-scent-biosynthesis-related lncRNAs, which highlights their potential functions in regulating the floral scent formation of jasmine and lays the foundations for future molecular breeding.

Ester based on l-menthol and phenibut ‒ (1R,2S,5R)-5-methyl-2-(propan-2-yl)cyclohexyl 4-amino-3-phenylbutanoate hydrochloride was obtained in 78% yield using N,N′-dicyclohexylcarbodiimide as a coupling reagent along with catalytic amount of 4-dimethylaminopyridine. The obtained product was characterized by FT-IR, fast-atom bombardment-mass spectrometry along with 1H-NMR and 13C-NMR spectral analysis; the purity was assessed using high-performance liquid chromatography. Phenibut ester has been examined on the models of chemically- and electrically-induced seizures for potential anticonvulsant profile.

Glechoma hederacea var. longituba (common name: ground ivy) has been used for the treatment of asthma, bronchitis, cholelithiasis, colds, and inflammation. In the present study, three new sesquiterpene glycosides (1–3), two new diterpene glycosides (4 and 5), and four known compounds (6–9) were isolated from its MeOH extract. A structure elucidation was performed for the five new compounds (1–5) using 1D and 2D NMR, HRESIMS, DP4+ and ECD calculations, and chemical methods. All the isolates (1–9) were assessed for their antineuroinflammatory activities on nitric oxide (NO) production in lipopolysaccharide (LPS)-activated BV-2 cells, nerve growth factor (NGF) secretion stimulation activities in C6 glioma cells, and cytotoxic activities against four human cancer cell lines (A549, SK-OV-3, SK-MEL-2, and HCT15). Compounds 2 and 5–7 exhibited inhibitory effects on the NO production with IC50 values of 52.21, 47.90, 61.61, and 25.35 μM, respectively. Compound 5 also exhibited a significant stimulating effect on NGF secretion (122.77 ± 8.10%). Compound 9 showed potent cytotoxic activity against SK-OV-3 (IC50 = 3.76 μM) and SK-MEL-2 (IC50 = 1.48 μM) cell lines, while 7 displayed a strong cytotoxic activity against the SK-MEL-2 (IC50 = 9.81 μM) cell line.

Lavender (Lavandula angustifolia Mill.) flowers and its distilled products are of major raw materials for cosmetic, perfumery, medicine and cleaning industries. Lavender flower production, essential oils and their qualities are affected by growing conditions and used agricultural methods. Mulching is a cultural application used for horticultural production in order to combat weeds, keep the soil temperature constant, eliminate soil moisture evaporation, provide favourable growth conditions for ample flower production, and improve the quality essential oil production. In this sense, plastic mulch application is the mainly preferred method by the growers, due to its low costs and durability. In the present study, two-year field (2014-2015) studies were carried out to evaluate the lavender flower production, essential oil yield in both conventional (without mulch) and plastic mulching systems in city of Kilis-Turkey. The individual essential oil compounds were identified using Gas Chromatography-Mass Spectrometer (GC-MS). Essential oils obtained from dried flowers were assayed for their neuroprotective effects. According to the results, fresh and dry inflorescence yields, dry flower yields, flower essential oil contents and compositions were affected by different growing methods, including (1) plastic mulching system and (2) conventional (without mulch) system. Considering the fresh inflorescence changes, mulching increased the yield by 147.84% in the first year while no significant changes were noted between treatments. Interestingly, dry inflorescence yield did not show a significant increase for both growing systems at first year but mulching enhanced the yield by 9.87% in the second year, in related to the traditional treatment. The results revealed that dry flower was not affected by both growing systems corresponding to the years. As for essential oil content, no changes were noted for both growing systems in the first year, whilst mulching increased the content by 13.73% in the second year. As the case of agricultural yield, the neuroprotective activities of essential oils obtained from mulching conditions exerted higher inhibitory roles against Acetyl-cholinesterase (AChE), butyryl-cholinesterase (BChE), and tyrosinase (TYRO) enzymes, in relative to the traditional treatments by 81.62%, 84.12%, 73.34% in the first year and by 76.15%, 79.80% and 68.02% in the second year. Such a high variation in neuroprotective activities can be explained by changes in essential oil contents, as observed for the percentage of the compounds. Consequently, these findings reveal that mulching application significantly contributes to the increase of agricultural yield and the growing of medicinal and aromatic plants that are more resistant to changing environmental conditions and biotic stress factors.

The rare edible and medicinal fungus Antrodia cinnamomea has a substantial potential for development. In this study, Illumina HiSeq 2000 was used to sequence its transcriptome. The results were assembled de novo, and 66,589 unigenes with an N50 of 4413 bp were obtained. Compared with public databases, 6,061, 3,257, and 2,807 unigenes were annotated to the Non-Redundant, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes databases, respectively. The genes related to terpene biosynthesis in the mycelia of A. cinnamomea were analyzed, and acetyl CoA synthase (ACS2 and ACS4), hydroxymethylglutaryl CoA reductase (HMGR), farnesyl transferase (FTase), and squalene synthase (SQS) were found to be upregulated in XZJ (twig of C. camphora) and NZJ (twig of C. kanehirae). Moreover, ACS5 and 2,3-oxidized squalene cyclase (OCS) were highly expressed in NZJ, while heme IX farnesyl transferase (IX-FIT) and ACS3 were significantly expressed in XZJ. The differential expression of ACS1, ACS2, HMGR, IX-FIT, SQS, and OCS was confirmed by real-time quantitative reverse transcription PCR. This study provides a new concept for the additional exploration of the molecular regulatory mechanism of terpenoid biosynthesis and data for the biotechnology of terpenoid production.

Abstract Background Productivities of bioactive compounds in high-value herbs and medicinal plants are often compromised by uncontrollable environmental parameters. Recent advances in the development of plant factories with artificial lighting (PFAL) have led to improved qualitative and/or quantitative production of bioactive compounds in several medicinal plants. However, information concerning the effect of light qualities on plant pharmaceutical properties is limited. The influence of three different light-emitting diode (LED) spectra on leaf fresh weight (FW), bioactive compound production and bioactivity of Artemisia annua L. against the malarial parasite Plasmodium falciparum NF54 was investigated. Correlation between the A. annua metabolites and antimalarial activity of light-treated plant extracts were also determined. Results Artemisia annua plants grown under white and blue spectra that intersected at 445 nm exhibited higher leaf FW and increased amounts of artemisinin and artemisinic acid, with enhanced production of several terpenoids displaying a variety of pharmacological activities. Conversely, the red spectrum led to diminished production of bioactive compounds and a distinct metabolite profile compared with other wavelengths. Crude extracts obtained from white and blue spectral treatments exhibited 2 times higher anti-Plasmodium falciparum activity than those subjected to the red treatment. Highest bioactivity was 4 times greater than those obtained from greenhouse-grown plants. Hierarchical cluster analysis (HCA) revealed a strong correlation between levels of several terpenoids and antimalarial activity, suggesting that these compounds might be involved in increasing antimalarial activity. Conclusions Results demonstrated a strategy to overcome the limitation of A. annua cultivation in Bangkok, Thailand. A specific LED spectrum that operated in a PFAL system promoted the accumulation of some useful phytochemicals in A. annua, leading to increased antimalarial activity. Therefore, the application of PFAL with appropriate light spectra showed promise as an alternative method for industrial production of A. annua or other useful medicinal plants with minimal environmental influence.

Having a spiral grain is considered to be one of the most important wood properties influencing wood quality. Here, transcriptome profiles and metabolome data were analyzed in the straight grain and twist grain of Pinus yunnanensis. A total of 6644 differential expression genes were found between the straight type and the twist type. A total of 126 differentially accumulated metabolites were detected. There were 24 common differential pathways identified from the transcriptome and metabolome, and these pathways were mainly annotated in ABC transporters, arginine and proline metabolism, flavonoid biosynthesis, isoquinoline alkaloid biosynthesis, linoleic acid metabolism, phenylpropanoid, tryptophan metabolism, etc. A weighted gene coexpression network analysis showed that the lightblue4 module was significantly correlated with 2′-deoxyuridine and that transcription factors (basic leucine zipper (bZIP), homeodomain leucine zipper (HD-ZIP), basic helix–loop–helix (bHLH), p-coumarate 3-hydroxylase (C3H), and N-acetylcysteine (NAC)) play important roles in regulating 2′-deoxyuridine, which may be involved in the formation of spiral grains. Meanwhile, the signal transduction of hormones may be related to spiral grain, as previously reported. ARF7 and MKK4_5, as indoleacetic acid (IAA)- and ethylene (ET)-related receptors, may explain the contribution of plant hormones in spiral grain. This study provided useful information on spiral grain in P. yunnanensis by transcriptome and metabolome analyses and could lay the foundation for future molecular breeding.

Sambiloto (Andrographis paniculata) is widely used as a medicinal plant for Indonesians. The content of extracted bioactive compounds is affected by the polarity of the extraction solvent. This study aims to determine the effect of extraction solvent on the chromatographic profile of Sambiloto herb, especially terpenoid group using the TLC-Spectrophotodensitometry method. Sambiloto herb was extracted by sonication method using methanol, ethanol, ethyl acetate, and chloroform as the solvents. Chromatographic profiles were identified by stationary phase of TLC silica gel GF254 plate and a mobile phase of chloroform:methanol (10:1 v/v), observed visually, and scanned at 210 nm. The plate was sprayed with anisadehyd-sulfuric acid reagent and observed visually. The compounds that belong to terpenoid compounds, namely S2, S5, S8, S11, and S12, had a greater percentage of peak area than the total area so that they were categorized as major components. The extraction solvent for Sambiloto herb that provides the best chromatographic profiles was found in the solvent of methanol extraction.

One of the local honey of South Sulawesi is Trigona honey which comes from Trigona spp. There are three types of trigona bees in South Sulawesi with each colony consisting of 300-80,000 individuals. Trigona bees are found nesting in hollow places such as logs, tree holes, and cracks in the walls of houses. The nest entrance is made of plant resin mixed with soil and mud, with different shapes and colors depending on the species. This type of bee produces a lot of propolis compared to honey and the price of honey produced is much higher because it has a chemical content (health benefits) that is much better than ordinary honey from the Apis mellifera group. This study was designed by extracting in ethanolic process at a concentration of 70%. The choice of concentration was based on previous research with an extraction temperature of 60 °C. The treatment of this research is to vary the time, namely 10 minutes, 20 minutes, and 30 minutes as a setting for the extraction time. Each treatment will be carried out in duplicate. This shows that in the honeycomb waste Trigona spp. contains terpenoids which are classified as antioxidants. Therefore, beehive waste Trigona spp. potential to be used as a source of antioxidants. In addition, several compounds are also relatively large antibacterial and anti-microbial compounds contained in the waste of Trigona spp beehives. The optimum solvent used by comparing the percentage of abundance (abundance/relative) through the percentage of the area and molecular stability after fragmentation is through the ethanolic process using ethanol solvent with percent abundance at 218 m/z and 189 m/z with the largest compound contained is Methyl Commate A and Alpha-Amyrin

IntroductionThe monoterpenoid linalool and sesquiterpenoid costunolide are ubiquitous plant components that have been economically exploited for their respective essential oils and pharmaceutical benefits. In general, monoterpenes and sesquiterpenes are produced by the plastid 2-C-methyl-D-erythritol 4-phosphate (MEP) and cytosolic mevalonate (MVA) pathways, respectively. Herein, we investigated the individual and combinatorial potential of MEP and MVA pathway genes in increasing linalool and costunolide production in Nicotiana benthamiana.MethodsFirst, six genes from the MEP (1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose 5-phosphate reductoisomerase, 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase, geranyl pyrophosphate synthase, and linalool synthase) and MVA (acetoacetyl-CoA-thiolase, hydroxy-3-methylglutaryl-CoA reductase, farnesyl pyrophosphate synthase, germacrene A synthase, germacrene A oxidase, and costunolide synthase) pathways were separately cloned into the modular cloning (MoClo) golden gateway cassette. Second, the cassettes were transformed individually or in combination into the leaves of N. benthamiana by agroinfiltration.Results and discussionFive days post infiltration (DPI), all selected genes were transiently 5- to 94-fold overexpressed. Quantification using gas chromatography-Q-orbitrap-mass spectrometry (GC-Q-Orbitrap-MS) determined that the individual and combinatorial expression of MEP genes increased linalool production up to 50–90ng.mg-1 fresh leaf weight. Likewise, MVA genes increased costunolide production up to 70–90ng.mg-1 fresh leaf weight. Our findings highlight that the transient expression of MEP and MVA pathway genes (individually or in combination) enhances linalool and costunolide production in plants.

Dioxinone derivatives, a class of acetoacetate derivatives, have attracted widespread attention because of their multiple reactive sites, high reactivity, unique chemical properties, and potential synthetic applications. The dioxinone group is also stable under a wide range of reaction conditions, including strong acids, as well as a variety of transition-metal-catalysed processes, such as olefin metathesis and Pd-mediated cross-coupling. The inherent reactivity and diverse applications of dioxinones make them valuable reactive intermediates in organic synthesis. The conversion of dioxinones to acylketenes and their subsequent nucleophilic capture is also an excellent strategy for synthesising β-keto acid derivatives, which can be applied even in complex molecular synthesis. This review focuses on the recent advances in the application of dioxinones in synthetic method research and the total synthesis of natural products, highlighting the exceptional utility of these synthetic methodologies in the construction of macrocyclic cores and terpenoid skeletons. In particular, successful transformations of dioxinone fragments are discussed.

Type 2 diabetes mellitus (T2DM) is a metabolic disease with persistent hyperglycemia primarily caused by insulin resistance (IR). The number of diabetic patients globally has been rising over the past decades. Although significant progress has been made in treating diabetes mellitus (DM), existing clinical drugs for diabetes can no longer fully meet patients when they face complex and huge clinical treatment needs. As a traditional and effective medical system, traditional Chinese medicine (TCM) has a unique understanding of diabetes treatment and has developed many classic and practical prescriptions targeting DM. With modern medicine and pharmacy advancements, researchers have discovered that various bioactive metabolites isolated from TCM show therapeutic on DM. Compared with existing clinical drugs, these bioactive metabolites demonstrate promising prospects for treating DM due to their excellent biocompatibility and fewer adverse reactions. Accordingly, these valuable metabolites have attracted the interest of researchers worldwide. Despite the abundance of research works and specialized-topic reviews published over the past years, there is a lack of updated and systematic reviews concerning this fast-growing field. Therefore, in this review, we summarized the bioactive metabolites derived from TCM with the potential treatment of T2DM by searching several authoritative databases such as PubMed, Web of Science, Wiley Online Library, and Springer Link. For the convenience of readers, the content is divided into four parts according to the structural characteristics of these valuable compounds (flavonoids, terpenoids, alkaloids, and others). Meanwhile, the detailed mechanism and future directions of these promising compounds curing DM are also summarized in the related sections. We hope this review inspires increasingly valuable and significant research focusing on potential bioactive metabolites from TCM to treat DM in the future.

InroductionVolatile organic compounds (VOCs) emitted by the receptive syconia of Ficus species is a key trait to attract their obligate pollinating fig wasps. Ficus hirta Vahl is a dioecious shrub, which is pollinated by a highly specialized symbiotic pollinator in southern China. Terpenoids are the main components of VOCs in F. hirta and play ecological roles in pollinator attraction, allelopathy, and plant defense. However, it remains unclear that what molecular mechanism difference in terpenoid synthesis pathways between pre-receptive stage (A-phase) and receptive stage (B-phase) of F. hirta syconia.MethodsTranscriptome, proteome and Gas Chromatography-Mass Spectrometer (GC-MS) were applied here to analyze these difference.Results and discussion:Compared to A-phase syconia, the genes (ACAT2, HMGR3, GGPS2, HDR, GPS2, TPS2, TPS4, TPS10-4, TPS14) related to the terpenoid synthesis pathway had higher expression level in receptive syconia (B-phase) according to transcriptome sequencing. Seven differentially expressed transcription factors were screened, namely bHLH7, MYB1R1, PRE6, AIL1, RF2b, ANT, VRN1. Specifically, bHLH7 was only specifically expressed in B-phase. 235 differentially expressed proteins (DEPs) were mainly located in the cytoplasm and chloroplasts. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the DEPs were mainly enriched in the metabolic process. A total of 9 terpenoid synthesis proteins were identified in the proteome. Among them, 4 proteins in methylerythritol phosphate (MEP) pathway were all down-regulated. Results suggested the synthesis of terpenoids precursors in B-phase bracts were mainly accomplished through the mevalonic acid (MVA) pathway in cytoplasm. Correlation analysis between the transcriptome and proteome, we detected a total of 1082 transcripts/proteins, three of which are related to stress. From the VOCs analysis, the average percent of monoterpenoids emitted by A-phase and B-phase syconia were 8.29% and 37.08%, while those of sesquiterpenes were 88.43% and 55.02% respectively. Monoterpenes (camphene, myrcene, camphor, menthol) were only detected in VOCs of B-phase syconia. To attract pollinators, B-phase syconia of F. hirta need more monoterpenoids and less sesquiterpenes. We speculate that transcription factor bHLH7 may regulate the terpenoid synthesis pathway between A- and B-phase syconia. Our research provided the first global analysis of mechanism differences of terpenoid synthesis pathways between A and B phases in F. hirta syconia.

Five new lipids, tricholixins A–E (1–5), and two known terpenoids, brasilane A (6) and harzianone A (7), were discovered from a deep-sea strain (R22) of the fungus Trichoderma lixii isolated from the cold seep sediments of the South China Sea. Their structures and relative configurations were identified by meticulous analysis of MS and IR as well as NMR data. The absolute configuration of 5 was ascertained by dimolybdenum-induced ECD data in particular. Compounds 1 and 2 represent the only two new butenolides from marine-derived Trichoderma, and they further add to the structural diversity of these molecules. Although 6 has been reported from a basidiomycete previously, it is the first brasilane aminoglycoside of Trichoderma origin. During the assay against wheat-pathogenic fungi, both 1 and 2 inhibited Fusarium graminearum with an MIC value of 25.0 μg/mL, and 6 suppressed Gaeumannomyces graminis with an MIC value of 12.5 μg/mL. Moreover, the three isolates also showed low toxicity to the brine shrimp Artemia salina.

Plants from the genus Styrax have been extensively used in folk medicines to treat diseases such as skin diseases and peptic ulcers and as an antiseptic and analgesic. Most Styrax species, especially Styrax tonkinensis, which is used as an expectorant, antiseptic, and analgesic in Chinese traditional medicine, could screen resin after external injury. Styrax is also used in folk medicines in Korea to treat sore throat, bronchitis, cough, expectoration, paralysis, laryngitis, and inflammation. Different parts of various Styrax species can be widely employed for ethnopharmacological applications. Moreover, for ethnopharmacological use, these parts of Styrax species can be applied in combination with other folk medicines. Styrax species consist of versatile natural compounds, with some of them exhibiting particularly excellent pharmacological activities, such as cytotoxic, acetylcholinesterase inhibitory, antioxidant, and antifungal activities. Altogether, these exciting results indicate that a comprehensive review of plants belonging to this genus is essential for helping researchers to continuously conduct an in-depth investigation. In this review, the traditional uses, phytochemistry, corresponding pharmacological activities, and structure–activity relationships of different Styrax species are clarified and critically discussed. More insights into potential opportunities for future research are carefully assessed.

Many species of the Dendrobium genus are traditional Chinese herbal medicine and ornamental plants. Flower fragrance is one of the most important horticultural ornamental characters and plays a crucial role in the ecology, economy, and aesthetics of plants. However, the volatile constituents and key regulatory genes related to floral biosynthesis are poorly understood. In this experiment, the flowers from two species of Dendrobium with high-scent smells, Dendrobium moniliforme (L.) Sw. (D. moniliforme), and light-scent smells, Dendrobium nobile “H1” (D. “H1”), were selected. The aim of this study was to explore the key gene expression profiles of floral biosynthesis by combining volatile constituent determination and transcriptome analysis in two different Dendrobium species. Physiological determination results showed that 60 volatile compounds were identified in D. moniliforme and 52 volatile compounds were identified in D. ‘H1’ flowers in four flowering stages, and the full bloom stage was the most complicated stage because there were 41 and 33 volatile compounds, respectively. These compounds belong to terpenes, aromatics, fatty acids, nitrogenous compounds, ketones, alcohols, and alkanes, respectively. The components identified in the gynandrium and petals revealed that the petals were probably the most important sites affecting the release of volatiles. The relative content of terpene compounds was the highest, with 77.25% (D. moniliforme) and 50.38% (D. “H1”), respectively. Transcriptome analysis showed that differentially expressed genes (DEGs) were highly enriched in terpenoid backbone biosynthesis and that four linalool synthetase (LIS) genes were up-regulated in high-scent smell species. This study is helpful to explore the key genes of flower fragrance and provides a theoretical basis for further understanding of the regulatory molecular functions of floral synthesis and release, as well as for the cultivation of new aromatic species.

The Oliveria decumbens Vent. is a wild, rare, annual medicinal plant and endemic plant of Iran that has metabolites (mostly terpenes) which make it a precious plant in Persian Traditional Medicine and also a potential chemotherapeutic agent. The lack of genetic resources has slowed the discovery of genes involved in the terpenes biosynthesis pathway. It is a wild relative of Daucus carota. In this research, we performed the transcriptomic differences between two samples, flower and root of Oliveria decumbens, and also analyze the expression value of the genes involved in terpenoid biosynthesis by RNA-seq and its essential oil’s phytochemicals analyzed by GC/MS. In total, 136,031,188 reads from two samples of flower and root have been produced. The result shows that the MEP pathway is mostly active in the flower and the MVA in the root. Three genes of GPP, FPPS, and GGPP that are the precursors in the synthesis of mono, di, and triterpenes are upregulated in root and 23 key genes were identified that are involved in the biosynthesis of terpenes. Three genes had the highest upregulation in the root including, and on the other hand, another three genes had the expression only in the flower. Meanwhile, 191 and 185 upregulated genes in the flower and root of the plant, respectively, were selected for the gene ontology analysis and reconstruction of co-expression networks. The current research is the first of its kind on Oliveria decumbens transcriptome and discussed 67 genes that have been deposited into the NCBI database. Collectively, the information obtained in this study unveils the new insights into characterizing the genetic blueprint of Oliveria decumbens Vent. which paved the way for medical/plant biotechnology and the pharmaceutical industry in the future.

Terpenoids are a diverse class of naturally occurring compounds consisting of more than 50,000 structurally different molecules and are found in all living organisms. Many terpenoid compounds, in particular those isolated from plants, have applications in various commercial sectors including medicine, agriculture and cosmetics. However, these high value terpenoids are produced in relatively small quantities in their natural hosts and their chemical synthesis for large scale production is costly and complicated. Therefore, there is much focus on producing these compounds in novel biological hosts using metabolic engineering technologies. As a photosynthetic system, the unicellular green alga C. reinhardtii is of particular interest as the most well-studied model alga with well-established molecular tools for genetic manipulation. However, the direct manipulation of terpenoid biosynthetic pathways in C. reinhardtii necessitates a thorough understanding of the basic terpenoid metabolism. To gain a better understanding of the methylerythritol phosphate (MEP) pathway that leads to terpenoid biosynthesis in the chloroplast of C. reinhardtii, hence this study has investigated the effect of over-expressing 1-deoxy-d-xylulose-5-phosphate synthase (DXS) on plastidic downstream terpenoids. We produced marker-free chloroplast transformants of C. reinhardtii lines that express an additional cyanobacterial gene for DXS. The analysis of terpenoid content for the transgenic line demonstrates that overexpressing DXS resulted in a two-fold decrease in the chlorophyll levels while carotenoid levels showed variable changes: zeaxanthin and antherxanthin levels increased several-fold, lutein levels dropped to approximately half, but β-carotene and violaxanthin did not show a significant change.

Floral terpenoid volatiles are impacted by light quality. In snapdragon, blue light can significantly enhance the emissions of ocimene and myrcene and the expression of ocimene synthase (AmOCS) and myrcene synthase (AmMYS). However, the mechanisms underlying the response to blue light are largely unknown. In this study, two transcription factors (TFs), AmMYB24 and AmMYB63 were screened which showed high expression level under blue light. AmMYB24 exhibited synchronous expression with AmOCS. Moreover, AmOCS transcript expression was up-regulated in response to AmMYB24 overexpression. This activation is direct and occurs through binding of AmMYB24 to MYBCORECYCATB1 sites in the AmOCS promoter. In addition, AmMYB24 interacts with the blue light signal key receptor AmCRY1 and the transcriptional activation activity of AmMYB24 was decreased in AmCRY1 silencing flowers. Taken together, our results revealed the regulatory pathway of biosynthesis of ocimene induced by blue light mediated by AmMYB24 and AmCRY1. When snapdragon flowers were exposed to blue light, AmCRY1 was first activated, the light signal is transduced to AmMYB24 through interaction with AmCRY1, and finally AmMYB24 activates AmOCS by binding to its MYBCOREATCYCB1 motif, resulting in abundant ocimene emission.

Synechococcus sp. PCC 7002 is a unicellular cyanobacterium capable of fast growth and tolerance to high light intensity and high salinity. These attributes along with genetic tractability make Synechococcus sp. PCC 7002 an attractive candidate for industrial scale production of specialty and commodity chemicals. Synechococcus sp. PCC 7002 LS (Davies et al., Front Bioeng Biotechnol, 2014, 2, 21–11) produces limonene, an energy dense diesel jet fuel drop-in additive, at a titer of 4 mg/L over a 4-day incubation period. In this study, we use the state-of-the-art whole-cell characterization tool, isotopically non-stationary 13C metabolic flux analysis (INST-13CMFA) to determine intracellular fluxes through the pathways of central metabolism for the limonene producing strain and wild type strain of Synechococcus sp. PCC 7002. We find similar flux distribution in the Calvin-Benson-Bassham cycle, photorespiration, oxidative pentose phosphate pathway, and oxidative tricarboxylic acid cycle. The key difference between strains is observed in the production of pyruvate. The limonene producing strain displays significantly higher flux through the amphibolic pathways of phosphoenolpyruvate carboxylase and the malic enzyme to synthesize pyruvate, while the wild type strain uses pyruvate kinase in a single step. Our findings suggest that this flux distribution is a mechanism to recover a physiologically optimal ratio of ATP to NADPH. The upregulation of this amphibolic pathway may act to restore the physiological ATP:NADPH ratio that has been disturbed by limonene biosynthesis. This study demonstrates the value of INST-13CMFA as a tool for cyanobacterial strain engineering and provides new avenues of research for improving limonene production in Synechococcus.

Carnosic acid (CA), a phenolic tricyclic diterpene, has many biological effects, including anti-inflammatory, anticancer, antiobesity, and antidiabetic activities. In this study, an efficient biosynthetic pathway was constructed to produce CA in Saccharomyces cerevisiae. First, the CA precursor miltiradiene was synthesized, after which the CA production strain was constructed by integrating the genes encoding cytochrome P450 enzymes (P450s) and cytochrome P450 reductase (CPR) SmCPR. The CA titer was further increased by the coexpression of CYP76AH1 and SmCPR ∼t28SpCytb5 fusion proteins and the overexpression of different catalases to detoxify the hydrogen peroxide (H2O2). Finally, engineering of the endoplasmic reticulum and cofactor supply increased the CA titer to 24.65 mg/L in shake flasks and 75.18 mg/L in 5 L fed-batch fermentation. This study demonstrates that the ability of engineered yeast cells to synthesize CA can be improved through metabolic engineering and synthetic biology strategies, providing a theoretical basis for microbial synthesis of other diterpenoids.

Elicitor treatments play an important role in inducing some protective signal transmitter enzymes in cells and regulating phenylalanine ammonia-lyase (PAL) activity. The aim of this study was to determine the effects of elicitors [silver nitrate (AgNO3), salicylic acid (SA) and yeast extract (YE)], which were added individually or in combination to Ocimum basilicum L. cell suspension cultures, on the production and antioxidant activity of secondary metabolites (SMs). Calluses were obtained from the leaves of O. basilicum kept on Murashige and Skoog (MS) medium containing 0.5 mg/l kinetin (KIN) + 2.5 mg/l naphthalene-acetic acid (NAA) and cell suspension cultures were initiated. Then elicitors were applied to the cell suspension cultures individually or in combination, and cells were harvested at the end of the second, fourth, and eighth days. Compared with the control culture, the maximum rosmarinic and chicoric acid production was obtained at the end of the 4th day from SA (24 µM) + YE (80 mg/l) treatment as 20.19 mg/g DW (118%) and 7.55 mg/g DW (123%), respectively. The maximum biosynthesis of isoquercetin and rutin compared with the control culture was 3.88 mg/g DW [YE (80 mg/l)] with a 1.6-fold increase and 6.35 mg/g DW [YE (80 mg/l) + AgNO3 (6 µM)] with a 1.76-fold increase, respectively. Estragole and linalool's highest values compared with the control culture were 4.50 μg/g DW [AgNO3 (6µM) + SA (24 µM)] and 3.02 μg/g DW [SA (24 µM)], respectively. Results clearly show that the elicitor treatment could enhance the biosynthesis of phenolic compounds and terpenoid content in cell suspension cultures of O. basilicum and may be used for commercial supply in the future for therapeutic applications.

Curcuma alismatifolia, a bulbous flower known for its showy bracts, is widely used around the world as a cut flower, potted, and garden plant. Besides its ornamental value, this species is rich in terpenoid metabolites and could serve as a resource for essential oils. Here, we report a chromosome-level genome assembly of C. alismatifolia and describe its biosynthetic pathways for anthocyanins and terpenoids. This high-quality, assembled genome size is 991.3 Mb with a scaffold N50 value of 56.7 Mb. Evolutionary analysis of the genome suggests that C. alismatifolia diverged from Zingiber officinale about 9.7 million years ago, after it underwent a whole-genome duplication. Transcriptome analysis was performed on bracts at five developmental stages. Nine highly expressed genes were identified, encoding for six enzymes downstream of the anthocyanin biosynthetic pathway. Of these, one gene encoding F3′5′H might be a key node in the regulation of bract color formation. Co-expression network analysis showed that MYB, bHLH, NAC, and ERF transcription factors collectively regulated color formation in the bracts. Characterization of terpenoid biosynthesis genes revealed their dispersal and tandem duplications, both of which contributed greatly to the increase in the number of terpene synthase genes in C. alismatifolia, especially to species-specific expansion of sesquiterpene synthase genes. This work facilitates understanding of genetic basis of anthocyanin and terpenoid biosynthesis and could accelerate the selective breeding of C. alismatifolia varieties with higher ornamental and medicinal value.

Several plant triterpenoids have valuable pharmaceutical properties, but their production and usage is limited since extraction from plants can burden natural resources, and result in low yields and purity. Here, we engineered oleaginous yeast Yarrowia lipolytica to produce three valuable plant triterpenoids (asiatic, madecassic, and arjunolic acids) by fermentation. First, we established the recombinant production of precursors, ursolic and oleanolic acids, by expressing plant enzymes in free or fused versions in a Y. lipolytica strain previously optimized for squalene production. Engineered strains produced up to 11.6 mg/g DCW ursolic acid or 10.2 mg/g DCW oleanolic acid. The biosynthetic pathway from ursolic acid was extended by expressing the Centella asiatica cytochrome P450 monoxygenases CaCYP716C11p, CaCYP714E19p, and CaCYP716E41p, resulting in the production of trace amounts of asiatic acid and 0.12 mg/g DCW madecassic acid. Expressing the same C. asiatica cytochromes P450 in oleanolic acid-producing strain resulted in the production of oleanane triterpenoids. Expression of CaCYP716C11p in the oleanolic acid-producing strain yielded 8.9 mg/g DCW maslinic acid. Further expression of a codon-optimized CaCYP714E19p resulted in 4.4 mg/g DCW arjunolic acid. Lastly, arjunolic acid production was increased to 9.1 mg/g DCW by swapping the N-terminal domain of CaCYP714E19p with the N-terminal domain from a Kalopanax septemlobus cytochrome P450. In summary, we have demonstrated the production of asiatic, madecassic, and arjunolic acids in a microbial cell factory. The strains and fermentation processes need to be further improved before the production of these molecules by fermentation can be industrialized.

Hyperlipidemia is a risk factor for various diseases, which is still a big problem in Indonesia. Yellow pumpkin (Cucurbita maxima D.) is a plant containing flavonoids and terpenoids, which can be used as an anti-hypercholesterolemic agent. This study aims to analyze the activity and dosage of yellow pumpkin extract, which can be used as an anti-hypercholesterolemic agent to reduce total cholesterol levels comparable to simvastatin by 1.8mg/kg BW/day. This research is pure experimental research with a pre and post-test group design approach. The number of samples was 25 male Wistar rats induced by high-fat feed, quail egg yolk: pork oil (2:1) by 5 ml/200 gramBW/day for 7 days. The extract dosage range was 200, 400, 600 mg/kg BW/ day for 7 days. Data were analyzed using One Way ANOVA. The results showed that the administration of yellow pumpkin extract could reduce rats' total blood cholesterol levels with a dose of 600 mg/kg BW/day, comparable to simvastatin 1.8 mg/kg BW/day. The secondary metabolites of the extract were flavonoids and terpenoids. Extract of yellow pumpkin (Cucurbita maxima D.) could reduce total blood cholesterol levels of rats. The dose of 600 mg/kg BW/day could reduce blood cholesterol levels in rats comparable to simvastatin 1.8 mg/kg BW/ day.

The WRKY gene family is one of the most significant transcription factor (TF) families in higher plants and participates in many secondary metabolic processes in plants. Litsea cubeba (Lour.) Person is an important woody oil plant that is high in terpenoids. However, no studies have been conducted to investigate the WRKY TFs that regulate the synthesis of terpene in L. cubeba. This paper provides a comprehensive genomic analysis of the LcWRKYs. In the L. cubeba genome, 64 LcWRKY genes were discovered. According to a comparative phylogenetic study with Arabidopsis thaliana, these L. cubeba WRKYs were divided into three groups. Some LcWRKY genes may have arisen from gene duplication, but the majority of LcWRKY evolution has been driven by segmental duplication events. Based on transcriptome data, a consistent expression pattern of LcWRKY17 and terpene synthase LcTPS42 was found at different stages of L. cubeba fruit development. Furthermore, the function of LcWRKY17 was verified by subcellular localization and transient overexpression, and overexpression of LcWRKY17 promotes monoterpene synthesis. Meanwhile, dual-Luciferase and yeast one-hybrid (Y1H) experiments showed that the LcWRKY17 transcription factor binds to W-box motifs of LcTPS42 and enhances its transcription. In conclusion, this research provided a fundamental framework for future functional analysis of the WRKY gene families, as well as breeding improvement and the regulation of secondary metabolism in L. cubeba.

Rosa rugosa is not only cultivated as a landscaping plant, but also used in cosmetics, the medical and food industries. However, little information is currently available on the gene regulatory networks involved in its scent and color biosynthesis and metabolism. In this study, R. rugosa Thunb. f. rosea Rehd with red petals (RR) and its white petal variant (WR), were used to study the molecular mechanisms in flower color and scent. Sixty-five differential flavonoid metabolites and 15 volatiles were found to have significant differences between RR and WR. Correspondingly, the key regulators (MYB-bHLH-WD40) of anthocyanin synthesis pathway and their structural genes involved in flavonoid biosynthesis, benzenoid/phenylpropanoid biosynthesis, terpenoid biosynthesis pathways were also found to be differentially expressed by comparative transcriptome. Further, qPCR permitted the identification of some transcripts encoding proteins that were putatively associated with scent and color biosynthesis in roses. Particularly, the results showed that the ACT gene (encoding CoA geraniol/citronellol acetyltransferase, GeneID: 112190420), which expressed lower in WR, was involved in three pathways: flavonoid biosynthesis, phenylpropanoid biosynthesis and terpenoid biosynthesis, however, GT5 (anthocyanin glycosylation gene, GeneID:112186660), expressed higher in WR, was involved in both flavonoid and phenylpropanoid biosynthesis pathways. These results suggested that ACT and GT5 might play important roles in regulating the relationship of color pigmentation and volatile emission.

The Mango parasite is a parasitic plant that is very detrimental to mango plant owners but has the potential as a producer of secondary metabolites which are expected to inhibit the growth of the fungus Candida albicans. This study aimed to obtain antifungal activity fractions, to obtain pure compound eluates which were active as antifungals, to determine the class of compounds active as antifungals, and to determine the Minimum Inhibitory Concentration (MIC) against the fungus C. albicans. Based on the research that has been done, the fractions that are active as antifungals are the n-hexane and ethyl acetate fractions. The results of purification of the n-hexane and ethyl acetate fractions using gravity column chromatography showed the presence of two pure n-hexane compounds that were active in inhibiting C. albicans, namely N-1 and N4 eluates with inhibition zone diameters of 12 mm and 10 mm, respectively. Purification of the ethyl acetate fraction produced two active compounds, namely eluate E-1 and eluate E-5 which were able to inhibit C. albicans by 10 mm and 12 mm, respectively. The MIC values ​​for N-1, N-4, and E-1 eluates were able to inhibit at a concentration of 500 ppm and were classified as quite strong. The E-5 eluate capable of inhibiting at a concentration of 1000 ppm is classified as weak. Tests for the class of compounds using Thin Layer Chromatography (TLC) and the R¦ value of the biautography results showed the color spots on the N-1 eluate (R¦: 0.9) were blue in the terpenoid group, on the N-4 eluate (R¦: 0.6) Alkaloid group is orange in color, the E-1 eluate (R¦: 0.8) is yellow in the Phenolic group, the E-5 eluate (R¦: 0.9) is purple in the terpenoid group.

Degraded limonoids are a subclass of limonoid natural products that derive from ring-intact or ring-rearranged limonoids. Establishment of robust synthetic routes to access them could provide valuable materials to identify the simplest active pharmacophore responsible for the observed biological activities of the parent molecules. This communication delineates the development of a divergent strategy to furnish melazolide B and several other related congeners from a common keto-lactone intermediate, which was rapidly assembled from α-ionone. A chemoselective carbonyl α,β-dehydrogenation and a Wharton reduction were key strategic steps in this synthetic pathway.

This article attempts to report native growth, plant description, phytochemical constituents and bioactivities of Syzygium aqueum, S. aromaticum, S. cumini, S. guineense and S. samarangense. Those are the large public species in the Syzygium genus and some of them have been used as traditional medicines. Different parts (leaves, seeds, fruits, barks, stem barks and flower buds) of each species plant are rich in phytochemical constituents such as flavonoids, terpenoids, tannins, glycosides and phenolics. Antioxidant, antidiabetic, anticancer, toxicity, antimicrobial, anti-inflammatory and anthelmintic activities are reported in various extracts (methanol, ethanol and aqueous) from different parts of Syzygium sp. The bioactivities were studied by using 1,1-diphenyl-2-picrylhydrazyl and ferric reducing antioxidant power assays for antioxidant, 5-(3-carboxymethoxyphenyl)-2-(4,5-dimethyl-thiazoly)-3-(4-sulfophenyl) tetrazolium and 3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assays for anticancer, α-glucosidase and α-amylase inhibition assays for antidiabetic, agar well diffusion method for antimicrobial and brine shrimp lethality assay for toxicity. Moreover, this review shows that phytochemical constituents of each species significantly presented various bioactivities. Therefore, this review suggests that there is great potential for obtaining the lead drug from these species.

Efficacy of fungi Volatile Organic Compounds. A review Introduction. Excessive use of chemical fungicides continues to drive research towards environmentally friendly and innovative alternatives for crop protection. Literature. Fungi produce various mixtures of compounds in the gas phase, called Volatile Organic Compounds (VOCs). They are able to diffuse into the soil and into the atmosphere and inhibit the activities of fungal pathogens. In this section, we will summarize recent knowledge on the inhibitory potential of Volatile Organic Compounds against pathogenic fungi with a focus on the effect of fungal VOCs. In practice, we will unveil initial research revealing their mode of action and any non-specific phytotoxic effects on the environmental microbiome and on plants. Conclusions. This article discusses new techniques used by researchers that focus on mycofumigation to optimize the formulation of a new generation of biofungicides. Thus, a new horizon is emerging for biological control of crop diseases.

Abstract Background Climate plays an essential role in forest health, and climate change may increase forest productivity losses due to abiotic and biotic stress. Increased temperature leads to the increased formation of ozone (O3). Ozone is formed by the interaction of sunlight, molecular oxygen and by the reactions of chemicals commonly found in industrial and automobile emissions such as nitrogen oxides and volatile organic compounds. Although it is well known that productivity of Northern red oak (Quercus rubra) (NRO), an ecologically and economically important species in the forests of eastern North America, is reduced by exposure to O3, limited information is available on its responses to exogenous stimuli at the level of gene expression. Results RNA sequencing yielded more than 323 million high-quality raw sequence reads. De novo assembly generated 52,662 unigenes, of which more than 42,000 sequences could be annotated through homology-based searches. A total of 4140 differential expressed genes (DEGs) were detected in response to O3 stress, as compared to their respective controls. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of the O3-response DEGs revealed perturbation of several biological pathways including energy, lipid, amino acid, carbohydrate and terpenoid metabolism as well as plant-pathogen interaction. Conclusion This study provides the first reference transcriptome for NRO and initial insights into the genomic responses of NRO to O3. Gene expression profiling reveals altered primary and secondary metabolism of NRO seedlings, including known defense responses such as terpenoid biosynthesis.