Search Results (196)

Background: Caryocar brasiliense Camb. Caryocaraceae is a typical tree from the Brazilian Cerrado with commercial importance due to its edible fruit, known as pequi. This native plant holds significant economic value and is a key candidate for cropping systems. Rich in phytochemicals, such as phenolics, flavonoids, and terpenoids, it has shown notable health benefits. Methods: Considering the importance of terpenes and their biological properties, and based on the first draft genome of C. brasiliense, this study aimed to identify putative terpene synthase genes and classify them into the phylogenetic subfamilies previously identified across all plant lineages. The presence of terpenes was also verified in samples of the outer portion of the fruit by solid-phase microextraction gas chromatography mass-spectrometry. Results: Analysis of genome completeness showed that over 90% of genes were identified despite a highly fragmented assembly, with 71% containing complete gene sequences. Twenty-two genes were retained as putative terpene synthase genes considering their homology with the terpene synthase Hidden Markov Model (HMM) profiles in the Pfam-A database. Ten sequences with a minimum length of 298 amino acids were used for phylogenetic inference. In the resulting phylogenetic tree, C. brasiliense terpene synthase genes clustered within the different previously identified Angiosperm clades and allowed us to classify each gene into different phylogenetic subfamilies: six genes belonged to the h/d/a/b/g, three to the c, and one to the e/f. The headspace solid-phase microextraction technique, in conjunction with gas chromatography mass-spectrometry, has allowed for the identification of eleven chemical compounds, including a terpene. Conclusions: This initial identification of putative terpene synthase genes in pequi, together with the chemical analysis of the outer fruits, lays the groundwork for future studies aimed at optimizing terpene biosynthesis for both biological and commercial applications. Full article

Terpenes are diverse plant metabolites with essential ecological and physiological functions, yet their biosynthetic regulation in lettuce (Lactuca sativa L.) remains poorly understood. By integrating volatile profiling, genome-wide identification, and biochemical characterization of terpene synthase (TPS) genes, we elucidated how methyl jasmonate (MJ) induces terpene formation in lettuce seedlings. Headspace analysis of 10-day-old seedlings revealed that while mock-treated tissues emitted no detectable volatiles, MJ elicitation triggered the de novo production of a terpene blend dominated by (E)-β-ocimene (9.3–14.6%), (E)-β-caryophyllene (37.2–46.9%), and caryophyllene oxide (26.2–41.4%). A genome-wide search identified 54 putative LsTPS genes, often clustered with prenyl transferases or cytochrome P450 genes. Gene expression assays revealed 17 MJ-responsive LsTPS genes; among them, LsTPS21, LsTPS23, LsTPS28, LsTPS51, and LsTPS52 showed strong (>200-fold) induction, with LsTPS52 exceeding a 20,000-fold increase. Functional characterization of six recombinant enzymes demonstrated diverse substrate specificities: LsTPS8 as an α-copaene synthase, LsTPS16 as a linalool synthase, LsTPS24 as an (E)-nerolidol synthase, LsTPS21 and LsTPS23 as (E)-β-ocimene synthases, and LsTPS10 as an (E)-β-caryophyllene synthase. Phylogenetic analyses confirmed conserved domains characteristic of the TPS-a and TPS-b subfamilies. This study presents the first comprehensive framework for MJ-induced terpene biosynthesis in lettuce, offering new insights into Asteraceae terpenoid metabolism. Full article

Floral volatile terpenoids are known to play important roles in plant pollination biology by attracting animal pollinators, repelling antagonists, and enhancing resistance to potential microbial pathogens. The terpenoid blend emitted by a flower is usually plant-lineage specific and is primarily determined by a set of versatile terpene synthases (TPSs), which catalyze the final step of diverse terpenoid synthesis. The strongly scented flower of Jasminum sambac (L.) Aiton emits linalool and α-farnesene, which dominate the nocturnal floral VOCs, yet the corresponding TPSs have not been identified. Here, we show that four TPS enzymes are responsible for the synthesis of a mixture of volatile terpenoids in the flower, based on their highly correlated and almost exclusive expression in the petal, as well as their enzymatic characterizations in vitro and in Nicotiana benthamiana Domin. JsTPS01 (TPS-a) acts as a sesquiterpene synthase, producing τ-cadinol in yeast at levels that mirror its rhythmic expression in petals. JsTPS02 (TPS-b) carries a plastid-targeting transit peptide, localizes to chloroplasts/plastids, and converts geranyl diphosphate (GPP) to linalool with high affinity (K_m = 28.2 ± 3.4 µM). JsTPS03 is a TPS-b clade member that can convert farnesyl diphosphate (FPP) to farnesol with a K_m of 14.4 ± 5.9 μM in an in vitro assay using isolated yeast vehicles. JsTPS04 (TPS-e/f) exhibits dual targeting—cytosolic in protoplasts of Arabidopsis thaliana (L.) Heynh, but plastidic in J. sambac petals—and functions as a bifunctional mono-/sesqui-TPS, forming linalool from GPP (K_m = 2.5 ± 0.3 µM) and trans-nerolidol from FPP (K_m = 7.6 ± 0.6 µM). Transient expression in N. benthamiana leaves further confirmed its in-planta linalool production. Collectively, we identified four preferentially expressed terpene synthases that contribute to the production of linalool, τ-cadinol, trans-nerolidol, and farnesol in J. sambac. Full article

Veronicastrum sibiricum (L.) Pennell, a species within the Plantaginaceae family, has a history of traditional application in addressing conditions such as abdominal pain, common cold, sore throat, parotitis, rheumatic discomfort, and snakebite. The plant produces diverse bioactive constituents, including phenylpropanoids, essential oils, flavonoids, and terpenoids. Terpenoids, generated via terpene synthases (TPSs), are of particular interest due to their pharmacological properties. Nevertheless, TPS enzymes in V. sibiricum have not been thoroughly investigated. In this research, a transcriptomic strategy was employed to isolate and profile TPS genes from V. sibiricum. Sequencing of the transcriptome produced 107,929 unigenes, among which 42,976 were functionally annotated using public databases. KEGG pathway examination revealed 264 genes associated with terpenoid metabolism, including 12 putative VsTPS genes harboring characteristic TPS domains. From these, VsTPS1 was successfully cloned. Functional characterization established that VsTPS1 operates as a bifunctional enzyme: in vitro, it catalyzes the conversion of FPP to (E)-nerolidol and, to a lesser extent, GPP to linalool. When expressed transiently in Nicotiana benthamiana, however, only (E)-nerolidol was detected, supporting its cytosolic localization and substrate specificity toward FPP. Accordingly, this sesquiterpene synthase was redesignated VsNES1. Co-expression of VsNES1 with HMGR in N. benthamiana markedly increased (E)-nerolidol yields, illustrating an effective strategy for heterologous production. These findings deepen our understanding of the TPS family in medicinal plant V. sibiricum and enable future biotechnological exploitation of terpenoid production in heterogenous plant cells. Full article

Perilla frutescens is an important medicinal and edible plant in Asia and was introduced in Europe and North America mainly as a spice plant. The commonly cultivated species is an allotetraploid (AABB). While the identity of its AA diploid donor has been preliminarily clarified, the other donor, BB, has not been discovered yet, and the taxonomic status and characteristics of the BB donor remain unresolved. Based on the published genomes of Perilla spp., we employed a collinearity analysis, gene structure similarity assessment, and multi-level functional annotation to infer the genomic and phenotypic features of the B subgenome. Results suggest that the protein sequences of the B and A subgenomes exhibit the highest similarity, while the protein sequences of Lavandula angustifolia or Ocimum basilicum are less similar to the B subgenome, and two subgenomes also possess the largest number of homologous genes and have similar gene structures. A total of 90 BB progenitor-specific genes were significantly enriched in pathways related to secondary metabolite biosynthesis and environmental stress response. Among these genes, the terpene synthase genes constitute the main genetic basis for the diversity of bioactive components in perilla. The discovery of a homologous gene containing the NB-ARC domain, associated with resistance to late blight, suggests that BB may contribute to key disease-resistant traits. Further gene family analysis revealed that compared with the A subgenome, the B subgenome exhibited fewer genes and lower diversity in the TPS and NB-ARC families. These findings indicate that BB may have originated from an unfound or extinct species within the Perilla spp. The BB donor might be less diversified than AA, possibly adapting to a narrow geographic and climatic range. Full article

The actinobacterial strain Kitasatospora griseola JNUCC 62 was isolated from volcanic wetland soil at Mulyeongari Oreum, Jeju Island, and taxonomically identified through 16S rRNA gene and whole-genome analyses. The complete genome, assembled from PacBio Sequel I reads, spans 8.31 Mb with a GC content of 72.8% and contains 7265 coding sequences. Comparative genomic indices (Average nucleotide identity, ANI 97.46%; digital DNA–DNA hybridization, dDDH 84.4%) confirmed its conspecific relationship with K. griseola JCM 3339^T. Genome mining using antiSMASH 8.0 revealed 30 biosynthetic gene clusters (BGCs), including polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS), ribosomally synthesized and post-translationally modified peptide (RiPP), lanthipeptide, and terpene types, accounting for 18.6% of the genome. Several BGCs displayed homology to known formicamycin-, lankacidin-, and lanthipeptide-type clusters, while others were novel or cryptic, reflecting adaptation to the nutrient-poor volcanic environment. Ethyl acetate extraction of the culture broth, especially under tryptophan-supplemented conditions, yielded four metabolites—1-acetyl-β-carboline, perlolyrine, tryptopol, and 1H-pyrrole-2-carboxylic acid—identified by UV and NMR spectroscopy. These compounds correspond to NRPS–PKS hybrid and arylpolyene-type gene clusters predicted in the genome, suggesting precursor-directed biosynthesis of indole and pyrrole alkaloids. The ethyl acetate extract (JNUCC62 EA) exhibited strong antioxidant capacity in the ABTS assay, anti-inflammatory activity via inhibition of nitric oxide (31.09 ± 3.69% of control) and cytokines (IL-6, IL-1β, TNF-α) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, and anti-melanogenic effects in α-melanocyte-stimulating hormone (MSH)-stimulated B16F10 melanoma cells, where melanin content and tyrosinase activity decreased to 61.49 ± 1.24% and 24.32 ± 0.31% of the control, respectively, without cytotoxicity. A human primary skin irritation test confirmed no irritation up to 50 µg/mL, establishing excellent dermal safety. Collectively, these findings highlight K. griseola JNUCC 62 from Mulyeongari Oreum as a volcanic wetland-derived actinomycete harboring rich biosynthetic potential for novel indole alkaloids with antioxidant, anti-inflammatory, and whitening properties, supporting its development as a safe and multifunctional cosmeceutical ingredient. Full article

Nilaparvata lugens is a migratory pest with high fecundity and outstanding drug resistance, which poses a devastating danger to rice production. This study investigated the reproductive regulation mechanism of N. lugens, specifically silencing the trehalose-6-phosphate synthase gene (TPS) via RNAi to elucidate how TPS governs the trehalose metabolic network through modulation of trehalose biosynthesis. Insect fecundity hinges on the synchronized progression of oogenesis and the tightly controlled expression of vitellogenin (Vg). In N. lugens, this coordination is orchestrated by an integrated molecular network that converges juvenile hormone signaling (JH), 20-hydroxyecdysone pathways (20E), insulin/IGF signaling (IIS), and the target of rapamycin cascade (TOR), collectively dictating the reproductive output of the species. Using TPS knockdown as the entry point, this study dissects the lipid-metabolic circuitry of N. lugens and uncovers how hormonal signaling cascades orchestrate reproduction by precisely modulating vitellogenin (Vg) and its cognate receptor VgR. Synthesized double-stranded terpene synthase genes (dsTPSs) can degrade mRNA, inhibit protein translation, and ultimately lead to the silencing of TPS genes, simultaneously crippling energy provision and hormonal signaling to orchestrate a multi-pronged suppression of reproduction. This dual-action intervention offers a promising molecular target for environmentally friendly management of N. lugens. Full article

Background: Asarum heterotropoides, a prominent medicinal plant in China, is well known for producing an abundance of monoterpenes and sesquiterpenes, which constitute the primary components of its essential oil and serve as the principal active compounds of the species. However, the biosynthetic pathways for these terpenoids remain largely unelucidated. Methods: Gas chromatography–mass spectrometry analysis, in vitro enzyme assay, subcellular localization experiment and molecular docking were used to characterize the function of terpene synthase from A. heterotropoides. Results: In this study, we isolated and characterized two monoterpene synthases and one sesquiterpene synthase from A. heterotropoides. These enzymes exhibit promiscuous activities, accepting geranyl pyrophosphate and farnesyl pyrophosphate as substrates to yield a variety of monoterpene and sesquiterpene products in in vitro enzymatic assays. All three enzymes possess a conserved RRx8W motif, a hallmark typically associated with TPS-b and TPS-d monoterpene synthases involved in cyclic monoterpene formation. However, these two monoterpene synthases yield linear instead of cyclic products. The sesquiterpene synthase (AhTPS3) is a second example of TPS-a terpene synthase harboring such motif. Conclusions: Our findings significantly expand our understanding of terpene biosynthesis, especially the role of RRx8W motif. Full article

Terpenes are the largest category of specialised metabolites. Aerobic endospore-forming bacteria (AEFB), a diverse group of microorganisms, can thrive in various habitats and produce specialised metabolites, including terpenes. This study investigates the potential for terpene biosynthesis in 10 AEFB strain whole-genome sequences by performing a bioinformatics analyses to identify genes associated with these isoprene biosynthesis pathways. Specifically, we focused on the sequences coding for enzymes in the methylerythritol-phosphate (MEP) pathway and the polyprenyl synthase family, which play crucial roles in synthesising terpene precursors together with terpene synthases. A comparative analysis revealed the unique genetic architecture of these biosynthetic gene clusters (BGCs). Our results indicated that some strains possessed the complete genetic machinery required to produce terpenes such as squalene, hopanoids, and carotenoids. We also reconstructed phylogenetic trees based on the amino acid sequences of terpene synthases, which aligned with the phylogenetic relationships inferred from the whole-genome sequences, suggesting that the production of terpenes is an ancestor property in AEFB. Our findings highlight the importance of genome mining as a powerful tool for discovering new biological activities. Furthermore, this research lays the groundwork for future investigations to enhance our understanding of terpene biosynthesis in AEFB and the potential applications of these Brazilian environmental strains. Full article

Terpenes, representing one of the most extensive classes of natural products, hold significant value in the fields of pharmaceuticals, fragrances, and biofuels. Extracting these compounds from natural sources is often environmentally unsustainable, and the structural diversity found in nature is inherently limited. Metabolic engineering using microbial hosts offers a scalable and sustainable alternative, utilizing optimized biosynthetic pathways—such as the mevalonate (MVA) and the methylerythritol phosphate (MEP) pathways—to achieve high-yield production of natural terpene scaffolds. This review focuses on the various strategies in developing microbial cell factories, ranging from enhancing precursor supply to optimizing terpene synthase systems. A new and promising frontier is the increase in structural diversity of terpenes by integration of non-biological chemical transformations into engineered biosynthetic pathways. We discuss the use of artificial metalloenzymes such as engineered cytochrome P450 variants that catalyze non-natural carbene transfer reactions (cyclopropanation). The merging of synthetic biology and synthetic chemistry goes beyond the normal synthesizing capabilities found in nature, which may pave the way for the design of “non-natural” terpenoids that contain new additions and better capabilities. Full article

A pathogenic fungus was isolated from the leaves of strawberry black spot in Zhengzhou China. Based on morphological and phylogenetic analysis, the isolate was identified as Alternaria alstroemeriae. Hybrid sequencing and assembly yielded a high-quality 38.7 Mb genome with 12,781 predicted genes and 99.6% Benchmarking Universal Single-Copy Orthologs (BUSCO) completeness. Functional annotation revealed enrichment in carbohydrate metabolism, secondary metabolite biosynthesis, and virulence-associated genes. Strain L6 harbored 45 biosynthetic gene clusters(BGCs), including 12 clusters for terpenes, 7 for non-ribosomal peptide synthetases, and 7 for polyketide synthases. Six BGCs showed high similarity to known pathways producing alternariol (phytotoxic/mycotoxic compound), alternapyrone (phytotoxin), choline (osmoprotectant), terpestacin (anti-angiogenic agent), clavaric acid (anticancer terpenoid), and betaenone derivatives (phytotoxins). CAZyme analysis identified 596 carbohydrate-active enzymes, aligning with L6’s biotrophic lifestyle. Additionally, 996 secreted proteins were predicted, of which five candidate effectors contained the conserved RxLx [EDQ] host-targeting motif, suggesting potential roles in virulence. This genome resource highlights L6’s exceptional secondary metabolites (SMs) diversity, featuring both plant-pathogenic toxins and pharmacologically valuable compounds, indicating that this endophytic fungus is a potential producer of metabolites meriting further exploration and development. Full article

Terpene synthases (TPS) facilitate terpenoid production, influencing the flavor, color, and medicinal properties of Crocus sativus (saffron), a triploid geophyte of significant commercial importance. Despite its importance, the CsTPS gene family remains poorly characterized, limiting genetic enhancements in saffron’s agronomic features. This research performed a comprehensive genome-wide analysis of CsTPS genes using genomic, transcriptomic, and in silico approaches. BLASTP and PfamScan discovered thirty CsTPS genes, demonstrating conserved TPS domains, varied exon–intron architectures, and chromosomal clustering indicative of tandem duplications. Phylogenetic research categorized these genes into five subfamilies (TPS-a to TPS-e), with the prevalence of TPS-a suggesting a role in sesquiterpene biosynthesis. RNA-seq data (PRJNA976833, PRJNA400472) revealed tissue-specific expression, with CsTPS1 and CsTPS5 expressed in reproductive tissues and CsTPS2 in vegetative tissues. Stress-responsive genes (CsTPS1, CsTPS4) exhibited upregulation in response to cold and pathogen stress, with cis-regulatory elements (e.g., ARE, ABRE) indicating hormone control. The in-silico validation of CsTPS1, chosen for its elevated GMQE score (0.89), included primer design, ePCR, and vector optimization for expression in Arabidopsis thaliana. This study elucidates the contribution of the CsTPS family to saffron terpenoid diversity, providing a foundation for enhancing flavor, yield, and stress tolerance through genetic engineering. Full article

Monoterpenoids serve as essential components of plant essential oils and play significant roles in plant growth, development, and insect resistance. Artemisia annua, an important medicinal plant, produces abundant terpenoids. While previous research on A. annua has predominantly focused on artemisinin biosynthesis and its regulation, studies on other terpenoids in this plant have significantly lagged behind. To comprehensively investigate monoterpene biosynthesis in A. annua, we analyzed monoterpenes across its different tissues using optimized extraction and chromatographic conditions developed to enhance sensitivity and resolution in our GC-MS-based analytical method. In A. annua, 31 monoterpenoid compounds were identified. Subsequently, eight candidate monoterpene synthases (mTPS) were characterized in Escherichia coli, confirming their catalytic activity in converting geranyl pyrophosphate (GPP) into distinct monoterpene products. Subcellular localization revealed these TPSs in chloroplasts, consistent with the widely reported chloroplast localization of TPS enzymes. These enzymes were functionally defined as monoterpenoid synthases, collectively responsible for synthesizing 18 monoterpenoid metabolites. Notably, AaTPS13, AaTPS19, and AaTPS20 exhibited substantial product promiscuity. Critically, the AaTPS19 was identified as the first known terpene synthase producing 2-pinanol. These findings systematically elucidate the biosynthesis of monoterpenoids in A. annua and provide key enzymatic elements for metabolic engineering and synthetic biology applications in monoterpenoid production. Full article

Terpenes, the largest class of plant specialized products, are built from C5 building blocks via terpene synthases and oxidized by cytochrome P450 enzymes (CYPs) for structural diversity. In some cases, CYPs do not simply oxidize the terpene backbone, but induce backbone rearrangements, methyl group shifts, and carbon–carbon (C–C) scissions. Some of these reactions were characterized over 25 years ago, but most of them were reported in recent years, indicating a highly dynamic research area. These reactions are involved in mono-, sesqui-, di- and triterpene metabolism and provide key catalytic steps in the biosynthesis of plant hormones, volatiles, and defense compounds. Many commercially relevant terpenoids require such reaction steps in their biosynthesis such as triptonide (rodent pest management), secoiridoids (flavor determinants), as well as ginkgolides, cardenolides, and sesquiterpene lactones with pharmaceutical potential. Here, we provide a comprehensive overview of the underlying mechanisms. Full article

Protected cultivation is a cultivation practice that plays an important role in improving crop quality. Aroma is an important flavour that assesses the quality of yuzu. In this study, C. junos cv. ‘Kitou’ grown in open fields (CJKTF) and plastic greenhouses (CJKTP) were selected as the study material. Significant differences in aroma performance between CJKTF and CJKTP were found by the olfactory senses of the members of this research group and an electronic nose, with CJKTP having a stronger aroma. Regarding VOCs, GC-MS analyses revealed 13 VOCs up-regulated and 28 VOCs down-regulated in CJKTP compared to CJKTF. Transcriptome analysis revealed that 515 genes were up-regulated and 720 genes were down-regulated in CJKTP compared to CJKTF. The differential VOCs nerolidol and γ-cadinene, and the differential genes nerolidol synthase 1 (NES1), nerolidol synthase 1-like (NES1-like), and cadinene synthase (DCS), were in the sesquiterpene synthesis pathway and showed significant correlation. NES1, NES1-like, and DCS encode terpene synthases, which may be involved in the biosynthetic pathway of nerolidol and γ-cadinene. In conclusion, the use of plastic greenhouses for cultivation may improve the quality and aroma intensity of yuzu, as well as alter the expression of related genes, compared to field cultivation. These results suggest that protected cultivation is a suitable cultivation practice to enhance the aroma of yuzu. Full article