Covering: 2012 to 2026Medicinal orchids constitute a rich yet vulnerable reservoir of high-value bioactive metabolites. However, their sustainable exploitation is severely hampered by resource depletion and the inefficiency of traditional extraction. Synthetic biology emerges as a transformative game-changer to resolve this impasse. This review delineates how synthetic biology drives a paradigm shift in endangered orchid conservation. Through modular design of biosynthetic pathways, heterologous reconstruction in microbial or plant chassis, directed evolution of key enzymes, metabolic engineering, and scalable production, it offers a disruptive solution, fundamentally redefining sustainable utilization by decoupling supply from ecological extraction. We highlight the successful heterologous biosynthesis of representative compounds like gastrodin and dendrobine. Facilitated by pathway optimization, chassis engineering and synthetic consortia design, these efforts have established cell factories that demonstrate the potential for sustainable and non-plant-based production, offering a viable alternative to wild harvest. Furthermore, the research encompasses recent breakthroughs in decoding orchid metabolic networks, including the elucidation of biosynthetic pathways, identification of core enzyme repertoires and discovery of regulatory switches involving transcription factors and miRNAs. Insights from evolutionary genomics and endophyte symbioses are also discussed as critical guides for pathway optimization. We explore the translational impact of these innovations, highlighting how the integration of multi-omics, artificial intelligence and gene editing accelerates the Design-Build-Test-Learn cycle. In summary, synthetic biology represents an essential paradigm shift, transitioning orchid resource management from a narrative of scarcity to one of predictable engineering creation and thereby securing a robust and sustainable supply chain for these prized medicinal plants.

We report the development of a novel nickel-hydride catalytic system for the reductive coupling of aryl halides with isoprene under mild conditions, allowing direct access to structurally diverse aryl isopentenyl compounds. The method requires no preformed metal reagents, exhibits excellent functional-group tolerance, overcomes regioselectivity challenges associated with isoprene, and provides a concise, efficient, and environmentally friendly route to natural and unnatural target products.

Drought stress severely threatens the productivity of Brassica napus. We revealed that two tandem CCCH zinc finger proteins, BnaTZF1A and BnaTZF1C, which are homologous to AtTZF1/2/3, function as crucial positive regulators of the drought response. Phenotypic, physiological, and cytological analyses revealed that overexpression of BnaTZF1A/1C improved the drought stress tolerance and led to late flowering, while knockdown of BnaTZF1A/1C dampened the drought tolerance. BnaTZF1A acts as an RNA-binding protein, binds to the 3'UTR stem-loop motifs of targets identified through RIP-seq and REMSA, including RALF and COP1-interactive protein, and is coregulated by BnaTZF1A and BnaTZF1C, resulting in depletion of target transcripts. BnaTZF1A also targeted BnaTZF1C mRNA and regulated its mRNA level to balance the stress signaling and fine-tune the drought stress response. Our results have unraveled the roles and possible regulatory mechanism of BnaTZF1 in response to drought stress, suggesting BnaTZF1A/1C as targets for improving drought stress tolerance in B. napus.

The cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) is a major pest of numerous crops. Plant responses to cotton bollworm attack are generally attributed to two primary factors: physical wounding and components of insect oral secretions. Discriminating between these responses is critical for understanding plant defense and developing pest control strategies. In this study, we found that cotton bollworm infestation specifically upregulated the expression of NtNAC29 and NtNAC94, whereas mechanical wounding did not. Functional analyses revealed that silencing either gene promoted bollworm growth, whereas overexpression suppressed larval development. These results demonstrated that NtNAC29 and NtNAC94 had pivotal roles in enhancing plant resistance to bollworm. Further analyses showed that both NAC transcription factors regulated the expression of Cysteine Protease Inhibitor 8 (NtCPI8) by binding to specific sites within the NtCPI8-1 and NtCPI8-2 promoters. Consistently, NtCPI8 exhibited a similar defensive role, significantly inhibiting cotton bollworm growth as reflected by reduced larval weight gain and shorter body length. Overall, our findings suggest that tobacco plants recognize cotton bollworm attack and activate downstream defense responses, including the induction of NtNAC29 and NtNAC94. These transcription factors in turn upregulate NtCPI8 expression, thereby strengthening plant resistance against insect herbivory. Notably, this NAC-CPI regulatory module is conserved among different crop species, providing a promising target for improving crop protection against herbivorous insects.

Benzylisoquinoline alkaloids (BIAs) represent a class of plant-derived compounds with significant pharmacological importance. The biosynthesis of most BIAs originates from a common precursor ( S)-reticuline. ( S)- N-methylcoclaurine 3'-hydroxylase (NMCH), encoded by CYP80B members, has always been regarded as a rate-limiting step that restricts the efficient synthesis of ( S)-reticuline, thereby leading to the low accumulation of BIAs. The highly-efficiency NMCH still needs to be further explored. Species within the Berberidaceae family are known to accumulate diverse BIAs at relatively high levels, making them an ideal plant resource for exploring highly active enzymes involved in BIA synthesis. Here, by integrating transcriptome and metabolite analysis across 11 Berberidaceae plants, we mine and characterize candidate genes involved in ( S)-reticuline biosynthesis. We further establish an engineered yeast platform for functional screening of NMCH genes and evaluate the catalytic activity of all CYP80B candidates. Among these, MbNMCH, isolated from Mahonia bealei, exhibits significantly higher catalytic activity for ( S)-reticuline production in yeast compared to previously reported NMCH enzymes. Our findings provide abundant genetic and metabolic information on Berberidaceae plants and identify a highly efficient enzymatic tool for BIA production in microbial cell factories, facilitating the sustainable manufacturing of diverse valuable alkaloids.

Although the anti-inflammatory effects of Baphicacanthus cusia polysaccharides have been partially validated, the structural features of these polysaccharides and their therapeutic roles in intestinal inflammation remain poorly understood. To address this gap, this study investigated the structural properties and therapeutic potential of BcP-b2, a novel polysaccharide isolated from Baphicacanthus cusia, in a dextran sulfate sodium (DSS)-induced colitis model. Structural analysis indicated that BcP-b2 is composed of arabinose, galactose, rhamnose, glucose, xylose, mannose, and fucose. Notably, BcP-b2 administration significantly alleviated colitis symptoms in mice, including weight loss, disease activity index score and histopathological damage in a concentration-dependent manner. Mechanistically, BcP-b2 enhanced intestinal barrier function by upregulating tight junction proteins Claudin-2 and Occludin, while reducing goblet cell depletion. Furthermore, BcP-b2 promoted beneficial shifts in the gut microbiota composition by increasing the abundance of short-chain fatty acid-producing bacteria and suppressing pathogenic genera, particularly Escherichia-Shigella. Integrated metabolomic analyses revealed concurrent improvements in bile acid homeostasis and butyrate metabolism, suggesting a coordinated therapeutic effect. Collectively, these findings underscore the potential of BcP-b2 as a promising candidate for the development of product aimed at treating and preventing colitis.

Septic cardiomyopathy is cardiac dysfunction caused by sepsis and is a common consequence of sepsis. Clinical studies have found patients with sepsis syndrome commonly have low serum selenium (Se) levels, and Se supplementation at doses higher than the daily requirement may reduce mortality; however, the underlying mechanism remains unclear. We found that downregulation of thioredoxin-2 (TXN2)-induced mitochondrial reactive oxygen species (mtROS), which increased inflammatory and oxidative injury in cardiomyocytes, might be a major contributor to septic cardiomyopathy. Mitoquinone mesylate (MitoQ), an antioxidant specifically targeted to mitochondria, increased TXN2 expression and decreased mtROS, thioredoxin-interacting protein (TXNIP, a negative regulator of TXN), nucleotide-binding oligomerization domain (NOD)-like receptor protein-3 (NLRP3) inflammasome activation, and ferroptosis. The results suggest that the TXN2-ferroptosis-NLRP3 pathway may represent a therapeutic target for sepsis-induced cardiac injury. In addition, we found that Se supplementation improved cardiac function and relieved mtROS accumulation and ferroptosis in sepsis-associated cardiac injury by regulating TXN2 and TXNIP/NLRP3 expression.

Medicinal plants are indispensable resources for traditional medical systems and natural product-based drug discovery. However, domestication and intensive breeding have reduced their genetic diversity and the complexity of their associated microbiomes, thereby limiting adaptability and stable production. Microbiome-based precision breeding provides a promising strategy for developing ideal medicinal plant (IPM) phenotypes characterized by high yield, high resistance, and high content of bioactive compounds. The composition, functions, and assembly mechanisms of plant-associated microbiomes are first summarized, followed by an evaluation of the current progress and limitations of precision agriculture microbiome engineering (PAME) in breeding programs. On this basis, a novel framework termed herb-microbiome interaction breeding (HMIB) is proposed. This framework emphasizes the mechanisms by which multilayer genetic regulation, including chromosomally encoded genes, cytoplasmic genetic factors, and microRNA-mediated regulation, shapes plant-associated microbiomes. Rational strategies for designing microbial inoculants are further discussed, including synthetic communities (SynComs), as well as the application potential of microalgae, nanomaterials, and prebiotics in constructing functional microbial inoculant systems. Finally, current challenges and future directions for the HMIB strategy in advancing the sustainable development of IPM are discussed.

Rosmarinic acid (RA) and salvianolic acid B (SAB), two major phenolic acid constituents of the medicinal plant Salvia miltiorrhiza, are widely used in pharmaceuticals and healthcare products. The biosynthesis of these compounds is influenced by various environmental factors, including UV-B radiation. Although the transcriptional regulation of phenolic acid biosynthesis by UV-B is relatively well understood, the molecular mechanisms underlying its post-translational regulation remain largely unexplored. In this study, we identified a UV-B-repressed Kelch repeat F-box protein gene, SmKFB2, which negatively regulates the accumulation of phenolic acids in S. miltiorrhiza. Proteomic analyses combined with western blot assays demonstrated that SmKFB2 modulates the endogenous protein levels of rosmarinic acid synthase (RAS). Furthermore, protein-protein interaction assays and co-expression studies revealed that SmKFB2 interacts with SmRAS and effects its stability. Additionally, UV-B was found to upregulate the gene expression of SmRAS. These findings suggest a dual regulatory mechanism: UV-B suppresses SmKFB2 expression, thereby reducing SmRAS degradation, which increases SmRAS protein stability and promotes RA and SAB accumulation; simultaneously, UV-B directly enhances SmRAS gene expression, further stimulating phenolic acid biosynthesis. Collectively, this study provides novel insights into the post-translational regulation of phenolic acid biosynthesis by UV-B and identifies potential targets for agricultural scientists in the development of new S. miltiorrhiza cultivars with enhanced medicinal value.

Background: Longhorn beetles, a widely recognized group of Chinese traditional medicinal insects, are characterized by their notable hemostatic properties. However, the comprehensive understanding of their medicinal potential has been hindered by the limitations of current research methodologies. Methods: This study focuses on the species Glenea cantor (Fabricius), which can produce several generations per year, and introduces a novel method using microwave carbonization techniques. By employing an in vitro coagulation test, UHPLC-MS, network pharmacology, molecular docking, and molecular dynamics simulation, the hemostatic efficacy and mechanism of action of Glenea cantor charcoal medicine (GC-CM) were thoroughly studied. Results: In vitro coagulation tests showed that GC-CM significantly reduced the activated partial thromboplastin time (APTT) and prothrombin time (PT), indicating its ability to enhance the coagulation cascade and preliminarily confirming its hemostatic efficacy (p < 0.01 vs. blank control group). The analysis revealed that GC-CM comprises 453 components, including 137 bioactive components with high human utilization. After predictions via databases such as SwissTargetPrediction and deduplication, 215 targets linked to hemostatic specificity were identified. These targets regulate signaling pathways such as platelet activation, complement and coagulation cascades, and cGMP-PKG. Molecular docking demonstrated strong affinities between key targets such as SRC and PIK3R1 and compounds such as 2′,6′-dihydroxy 4′-methoxydihydrochalcone, and 1-monolinoleoyl-rac-glycerol (binding energy < −5 kcal/mol). Molecular dynamics simulations show good binding capacity between core components and targets Conclusions: The aim of this study was to elucidate the material basis and mechanism of the hemostatic efficacy of GC-CM, offering a model for exploring other insect-based medicinal resources.

Indigo, a plant-originated blue dye, has a long and well-documented history of extensive human use. The Isatis genus has long been a key source for indigo production, however, the biosynthetic pathway responsible for indigo within Isatis has remained elusive. Here, we conducted phylogenetic and metabolic analyses of various Isatis taxa, revealing that the capacity to produce indigo was apparently lost in some of these taxa. Following de novo genome sequencing, assembly, and comparative genomic analysis between Isatis indigotica and Isatis cappadocica, we delved into the origins and evolution of indigo biosynthesis. Homologous expression of candidate genes in Nicotiana benthamiana identified multiple oxidase families, including flavin-containing monooxygenase (FMO) and cytochrome P450 (CYP) protein that catalyze the oxidation steps leading to the indigo biosynthesis, indicating a metabolic innovation derived from the oxime pathway in plants. The evolutionary aspects concerning the neofunctionalization of CYPs-catalyzed biosynthesis of glucosides and FMOs-catalyzed oxime in Isatis taxa provide new insights into the evolution of these metabolic pathways in plants.

Artemisinin and its derivatives are widely recognized for their exceptional antimalarial efficacy. Recently, accumulating evidence indicates therapeutic potential beyond malaria. Despite these advances, detailed mechanisms and pharmacological limitations remain incompletely defined. This review summarizes their pharmacological activities and molecular mechanisms associated with oncology, immunoregulation, and metabolic disorders. Mechanistically, these compounds exert potent antitumor effects by inducing oxidative stress, arresting the cell cycle, triggering apoptosis, and inhibiting angiogenesis. They likewise modulate immune responses, re-establishing immune homeostasis and enhancing the effectiveness of immunotherapeutic strategies. Preliminary evidence also suggests involvement in metabolic regulation, pointing to promising avenues for treating metabolic disorders. Given alternative mechanisms of artemisinin and its derivatives, we also discuss the trinity modulation network among antitumor activity, immunoregulation, and metabolic homeostasis. We anticipate that future research will address these knowledge gaps, thereby enhancing the clinical utility of artemisinin and its derivatives and improving patient outcomes across diverse pathologies.

Background Esophageal squamous cell carcinoma (ESCC) is a leading cause of cancer mortality globally, with pronounced geographic disparities in incidence. Emerging evidence links oral microbiome dysbiosis to ESCC pathogenesis, yet comprehensive insights into microbial diversity, taxonomic shifts, and functional alterations in high-risk populations remain limited. Methods Using 16S rRNA amplicon sequencing, we compared the oral microbiome of ESCC patients and healthy controls from a high-incidence region in Northwest China. Alpha and beta diversity metrics, taxonomic composition, and predicted functional pathways were analyzed to identify microbial signatures associated with ESCC. Results ESCC patients exhibited significantly elevated microbial richness (observed amplicon sequence variants (ASVs), Chao1, ACE; p < 0.05) but comparable Shannon/Simpson diversity to controls. Unique amplicon sequence variants (ASVs) were more prevalent in ESCC samples, and principal component analysis confirmed distinct community structures (p < 0.05). Taxonomically, Streptococcus and Neisseria dominated both groups, but ESCC patients showed enrichment of Gemella (p = 0.0003) and Corynebacterium (p < 0.00001), alongside depletion of Prevotella_7 (p = 0.0002) and Moraxella (p < 0.001). Functional profiling revealed upregulated amino acid metabolism (e.g., beta-alanine and valine degradation) and downregulated carbohydrate metabolism in ESCC-associated microbiota. Conclusion This study uncovers unique oral microbial signatures in ESCC patients from a high-incidence region, characterized by increased richness, taxon-specific shifts, and metabolic reprogramming favoring amino acid catabolism. These findings highlight the potential of microbial biomarkers for ESCC detection and provide mechanistic insights into microbiome-driven carcinogenesis. The geographic specificity of the cohort underscores the urgency of tailored interventions in high-risk populations and advances our understanding of microbial contributions to esophageal cancer.

Tanshinones (TAs), well-known specialized diterpenoid metabolites in Salvia plants, exhibit distinct tissue-specific production in the root periderm. However, the mechanisms regulating this accumulation pattern remain unknown. Here, we employed a multi-omics analysis strategy to uncover the transcriptional regulatory network responsible for TA biosynthesis in Salvia miltiorrhiza roots. By integrating metabolic profiling, RNA-seq, and ATAC-seq, we profile the temporo-spatial dynamics of metabolic, transcriptional, and chromatin landscapes during early root development. Our results demonstrate that TAs biosynthesis and accumulation in S. miltiorrhiza roots display spatiotemporal patterns, marked by periderm-specific accumulation and initiation exclusively at specific developmental stages, tightly coordinated with dynamic changes in chromatin accessibility and transcriptional regulation. The constructed transcriptional regulatory network driving TA biosynthesis was found to be dominated by 211 key transcription factors (TFs). Experimental validations highlighted SmERF105 as a key positive regulator of TA, activating the transcription of KSL1, CYP76AH3, and the TA transporter ABCG1 to modulate the TA production. Our study uncovers novel, high-confidence regulators and offers an effective strategy for dissecting the genetic basis of plant specialized metabolites, offering value for advancing TA metabolic engineering.

ETHNOPHARMACOLOGICAL RELEVANCE:Female breast cancer ranks second in incidence rate and fourth in mortality globally. Tylophora yunnanensis Schlechter (Asclepiadaceae) is frequently used in folk medicine to treat irregular menses, falls caused injuries, rheumatoid arthritis, hepatitis, gastric ulcers, and gynecological tumors. AIMS OF THE STUDY:To explore the molecular mechanism of T. yunnanensis against breast cancer. MATERIALS AND METHODS:Numerous experiments were implemented for detection of cell proliferation, death, toxicity, MMP, cycle, apoptosis, DNA damage, and cholesterol levels. Transcriptomic analysis, proteomic analysis, exogenous cholesterol antagonism, gene overexpression and Western blot were performed to explore the mechanism of action. A tumor-bearing animal model was utilized. The chemical composition of T. yunnanensis extract (TYE) was analyzed using LC-MS/MS. RESULTS:TYE repressed the proliferation of BT549 and 4T1 cells, with IC50 values of 4.88, 2.98 μg/mL for 24 h and 4.70, 1.87 μg/mL for 48 h, respectively. Cell cycle arrest was also induced. The multi-omics analysis displayed that TYE suppressed cholesterol biosynthesis and interfered with DNA damage repair in TNBC cells. Exogenous cholesterol reversed these effects, counteracted the elevated intracellular TC, FC, γ-H2AX, and tail moment. TYE downregulated the expression of SQLE, MVK, FDPS, TM7SF2, and DHCR24 proteins, which was offset by addition of cholesterol. Overexpression of SQLE reduced the inhibition effects of TYE on cell viability but was not responsible for the expression inhibition of MVK, FDPS and DHCR24 proteins. TYE decreased the tumor growth, serum TC and TG, and in situ expression of ki67 and SQLE in tumor-bearing mice and had no acute toxicity to mice. No chemical components of TYE were identified. CONCLUSION:TYE inhibits SQLE transcription to decrease its protein expression, reduces cholesterol biosynthesis and accumulation, impedes DNA damage repair, leading to cell cycle arrest, and thus elicits cell death, followed by obstruction of breast cancer progression. T. yunnanensis may be a novel anti-breast cancer agent owing to its inhibitory effects on cholesterol biosynthesis.

Baphicacanthus cusia (Nee) Bremek, a perennial herbaceous plant with medicinal properties, has limited genomic insights regarding the genes involved in its indole alkaloid biosynthesis pathway. In this study, the BcSK gene was isolated and cloned from the transcriptome data of B. cusia. The full-length cDNA of BcSK is 1,657 bp, comprising a 265 bp 5’ UTR, a 507 bp 3’ UTR, and an 885 bp ORF encoding 295 amino acids. The exon-intron structure of BcSK consists of four exons and three introns. Bioinformatics and phylogenetic analyses revealed a high degree of homology between BcSK and its counterparts in various plant species. Quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed that BcSK expression was significantly altered under abiotic stress conditions, including methyl jasmonate (MeJA), abscisic acid (ABA), and ultraviolet (UV) radiation. The gene was predominantly expressed in flowers compared to roots, stems, and leaves. Subcellular localization analysis indicated that BcSK is primarily expressed in chloroplasts, confirming that the conversion of shikimic acid to shikimate-3-phosphate occurs in this organelle. Prokaryotic expression and enzyme activity assays demonstrated that the heterologously expressed BcSK protein catalyzed the conversion of shikimic acid to shikimate-3-phosphate. Furthermore, the ectopic overexpression of BcSK in Isatis indigotica significantly enhanced the biosynthetic flux toward indole alkaloids, including indole, indigo, and indirubin. In conclusion, this study identifies and characterizes a novel BcSK gene, providing new insights and potential applications for the metabolic engineering of B. cusia.

Paclitaxel (PTX), a valuable natural product derived from Taxus species, exhibits remarkable anti-cancer properties. It penetrates nanopores in microtubule walls, interacting with tubulin on the lumen surface and disrupting microtubule dynamics, thereby inducing cytotoxic effects in cancer cells. PTX and its derivatives have gained approval for treating various diseases due to their low toxicity, high efficiency, and broad-spectrum application. The widespread success and expanding applications of PTX have led to increased demand, raising concerns about accessibility. Consequently, researchers globally have focused on developing alternative production methods and applying nanocarriers in PTX delivery systems to enhance bioavailability. This review examines the challenges and advancements in PTX sourcing, production, physicochemical properties, anti-cancer mechanisms, clinical applications, trials, and chemo-immunotherapy. It aims to provide a comprehensive reference for the rational development and effective utilization of PTX.