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Objectives: Light intensity is a critical environmental factor regulating plant growth, development, and stress adaptation. However, the physiological and molecular mechanisms underlying light responses in Aconitum kusnezoffii, a valuable alpine medicinal plant, remain poorly understood. This study aimed to elucidate the adaptive strategies of A. kusnezoffii under different light intensities through integrated physiological and transcriptomic analyses. Methods: Two-year-old A. kusnezoffii plants were exposed to three controlled light regimes (790, 620, and 450 lx). Leaf anatomical traits were assessed via histological sectioning and microscopic imaging. Antioxidant enzyme activities (CAT, POD, and SOD), membrane lipid peroxidation (MDA content), osmoregulatory substances, and carbon metabolites were quantified using standard biochemical assays. Transcriptomic profiling was conducted using Illumina RNA-seq, with differentially expressed genes (DEGs) identified through DESeq2 and functionally annotated via GO and KEGG enrichment analyses. Results: Moderate light (620 lx) promoted optimal leaf structure by enhancing palisade tissue development and epidermal thickening, while reducing membrane lipid peroxidation. Antioxidant defense capacity was elevated through higher CAT, POD, and SOD activities, alongside increased accumulation of soluble proteins, sugars, and starch. Transcriptomic analysis revealed DEGs enriched in photosynthesis, monoterpenoid biosynthesis, hormone signaling, and glutathione metabolism pathways. Key positive regulators (PHY and HY5) were upregulated, whereas negative regulators (COP1 and PIFs) were suppressed, collectively facilitating chloroplast development and photomorphogenesis. Trend analysis indicated a “down–up” gene expression pattern, with early suppression of stress-responsive genes followed by activation of photosynthetic and metabolic processes. Conclusions: A. kusnezoffii employs a coordinated, multi-level adaptation strategy under moderate light (620 lx), integrating leaf structural optimization, enhanced antioxidant defense, and dynamic transcriptomic reprogramming to maintain energy balance, redox homeostasis, and photomorphogenic flexibility. These findings provide a theoretical foundation for optimizing artificial cultivation and light management of alpine medicinal plants.

In this study, we conducted an assembly and analysis of the organelle genomes of Aconitum carmichaelii . Our investigation encompassed the examination of organelle genome structures, gene transfer events, and the environmental selection pressures affecting A. carmichaelii . The results revealed distinct evolutionary patterns in the organelle genomes of A. carmichaelii . Especially, the plastome exhibited a more conserved structure but a higher nucleotide substitution rate (NSR), while the mitogenome displayed a more complex structure with a slower NSR. Through homology analysis, we identified several instances of unidirectional protein-coding genes (PCGs) transferring from the plastome to the mitogenome. However, we did not observe any events which genes moved from the mitogenome to the plastome. Additionally, we observed multiple transposable element (TE) fragments in the organelle genomes, with both organelles showing different preferences for the type of nuclear TE insertion. Divergence time estimation suggested that rapid differentiation occurred in Aconitum species approximately 7.96 million years ago (Mya). This divergence might be associated with the reduction in CO 2 levels and the significant uplift of the Qinghai-Tibet Plateau (QTP) during the late Miocene. Selection pressure analysis indicated that the dN/dS values of both organelles were less than 1, suggested that organelle PCGs were subject to purification selection. However, we did not detect any positively selected genes (PSGs) in Subg. Aconitum and Subg. Lycoctonum . This observation further supports the idea that stronger negative selection pressure on organelle genes in Aconitum results in a more conserved amino acid sequence. In conclusion, this study contributes to a deeper understanding of organelle evolution in Aconitum species and provides a foundation for future research on the genetic mechanisms underlying the structure and function of the Aconitum plastome and mitogenome.

Aconitum kusnezoffii is an important medicinal plant whose seeds exhibit deep physiological dormancy. To elucidate the endogenous hormonal regulation mechanisms and dynamic gene expression patterns during dormancy release, we conducted a comprehensive analysis integrating hormone quantification and transcriptome sequencing on seed samples subjected to cold stratification for 0, 14, and 42 days. Morphological observations indicated that the embryo development rate remained stable during the first 14 days of stratification but increased significantly after 21 days, displaying a sigmoidal growth pattern. This provided a rationale for delineating the dormancy maintenance, release initiation, and pre-germination phases and for selecting key sampling time points. Hormonal profiling revealed a continuous decline in abscisic acid (ABA) content, accompanied by a gradual increase in germination-promoting hormones such as gibberellins (bioactive GA, reported as GA₃-equivalents), indole-3-acetic acid (IAA), brassinolide (BL) and methyl jasmonate (MeJA). The ratios of GA/ABA (GA₃-equivalents), IAA/ABA, and zeatin riboside (ZR; ELISA-based total tZR + cZR equivalents)/ABA increased accordingly, suggesting that dormancy release is orchestrated through a synergistic interplay of multiple hormones. Transcriptome analysis yielded 79,251 Unigenes, with differentially expressed genes significantly enriched in pathways related to energy metabolism, hormone signal transduction, and carbohydrate metabolism. STEM clustering identified five representative temporal expression profiles. Further analysis of hormone-related pathways showed that ABA biosynthesis and signaling components were predominantly active at early stages but declined thereafter, whereas GA and IAA biosynthetic and signaling genes were markedly upregulated during the later stages of dormancy release. These findings highlight the close association between embryo development and hormonal dynamics during seed dormancy release in A. kusnezoffii , and clarify the stage-specific features of hormone synthesis and signaling pathways, providing a theoretical basis for understanding dormancy mechanisms and improving artificial germination techniques.

Diterpenoids exhibit remarkable structural diversity and bioactivities, shaped primarily by the tandem actions of skeleton-forming terpene synthases (TPSs) and cytochrome P450 monooxygenases. The e nt -kaurene and ent -atiserene are labdane-derived diterpene scaffolds for the biosynthesis of diverse bioactive diterpenoids and diterpene alkaloids, including the clinically used analgesic 3-acetylaconitine and anti-arrhythmic guan-fu base A in Aconitum spp., yet what and how P450s drive their structural and functional diversification remain largely unexplored. Here, via mining the transcriptomes of Aconitum carmichaelii and Aconitum coreanum followed by functional validation, we discover nine TPSs capable of furnishing the ent -kaurene, ent -atiserene or 16 α -hydroxy- ent -kaurene scaffold and 14 divergent P450s, of which eight are multifunctional, catalyzing oxidation at seven different sites of the scaffolds. Protein analysis and mutagenesis experiments reveal key residues tuning the P450 activity and product profiles, shedding light on how they diverge functionally. The discovered TPSs and P450s enable combinatorial biosynthesis of tripterifordin, guan-fu diterpenoid A, and 14 novel atiserenoids, with some exhibiting allelopathic activity. Intriguingly, Tripterygium wilfordii -derived tripterifordin was detected in A. coreanum , revealing convergent biosynthesis of tripterifordin. Our findings highlight P450 plasticity in selective diterpene oxidation and allow for accessing tripterifordin and derivatives, paving ways for elucidating downstream diterpenoid pathways via synthetic biology. Medicinal Aconitum species produce diterpene alkaloids with diverse oxidation catalyzed by yet unknown enzymes. Luo et al. discover that divergent multifunctional P450s are key to driving oxidative diversification, yielding cryptic bioactive diterpenoids in Aconitum spp.

Aconitum stands out among the Ranunculaceae family for its notable use as an ornamental and medicinal plant. Diterpenoid alkaloids (DAs), the characteristic compounds of Aconitum, have been found to have effective analgesic and anti-inflammatory effects. Despite their medicinal potential, the toxicity of most DAs restricts the direct use of Aconitum in traditional medicine, necessitating complex processing before use. The use of high-throughput omics allows for the investigation of Aconitum plant genetics, gene regulation, metabolic pathways, and growth and development. We have collected comprehensive information on the omics studies of Aconitum medicinal plants, encompassing genomics, transcriptomics, metabolomics, proteomics, and microbiomics, from internationally recognized electronic scientific databases such as Web of Science, PubMed, and CNKI. In light of this, we identified research gaps and proposed potential areas and key objectives for Aconitum omics research, aiming to establish a framework for quality improvement, molecular breeding, and a deeper understanding of specialized metabolite production in Aconitum plants.

Background Aconitum transsectum Diels. (Ranunculaceae) is an important medicinal plant that is widely used in traditional Chinese medicine, but its morphological traits make it difficult to recognize from other Aconitum species. No research has sequenced the chloroplast genome of A.transsectum , despite the fact that phylogenetic analysis based on chloroplast genome sequences provides essential evidence for plant classification . Results In this study, the chloroplast (cp) genome of A. transsectum was sequenced, assembled, and annotated. A. transsectum cp genome is a 155,872 bp tetrameric structure including a large single copy (LSC, 87,671 bp) and a small single copy (SSC, 18,891 bp) section, as well as a pair of inverted repeat sequences (IRa and IRb, 25,894 bp each). 131 genes are encoded by the complete cp genome, comprising 86 protein-coding genes, 37 tRNAs, and 8 rRNAs. The most favored codon in the A. transsectum cp genome is AUG, and 46 repeats and 241 SSRs were also identified. The A. transsectum cp genome is similar in size, gene composition, and IR expansion and contraction to the cp genomes of seven Ranunculaceae species. Phylogenetic analysis of cp genomes of 28 plants from the Ranunculaceae family shows that A. transsectum is most closely related to A. vilmorinianum , A . episcopale , and A. forrestii of Subgen. Aconitum . Conclusions Overall, this study provides complete cp genome resources for A. transsectum that will be beneficial for identifying potential.

Background Aconitum is an important medicinal genus widely used in traditional Chinese medicine, which produces types of diterpenoid alkaloids (DA) among different species. We performed whole genome resequencing (WGS) research in Aconitum spp., and wish to find diterpenoid alkaloids related genetic variations. Results In this study, we re-sequenced 150 Aconitum vilmorinianum (A. vilmorinianum) including 102 from the cultivation garden and 48 from the wild, as well as nine wild samples of Aconitum weixiense. The intra-population differentiation of A. vilmorinianum was detected by evolutionary tree and population structure inference. We identify 47 DA biosynthesis genes that might be highly associated with the specialization of DA based on whole-genome resequencing. Of 616 significant SNPs and 105 significant InDels among these genes could be developed as polymorphic molecular markers capable of effectively recognizing A. vilmorinianum from A. weixiense . Furthermore, the significant SNPs and InDels were almost homozygous alternates in A. weixiense , whereas they tended to be homozygous references in the A. vilmorinianum . Conclusions Our results discussed the difference in genetic background in A. vilmorinianum compared to A. weixiense and these high-quality DA biosynthesis-associated polymorphic locus provided useful genetic information for discrimination of A. vilmorinianum and could serve as a vehicle to study the mechanism of DA differentiation in Aconitum .

Chitosan (CTS) is recognized for enhancing a plant’s resilience to various environmental stresses, such as salinity and drought. Moreover, salicylic acid (SA) is acknowledged as a growth regulator involved in addressing metal toxicity. However, the effectiveness of both compounds in mitigating Cr-induced stress has remained relatively unexplored, especially in the case of Aconitum napellus , a medicinally and floricultural important plant. Therefore, the primary objective of this study was to investigate the potential of CTS and SA in alleviating chromium (Cr)-induced stress in A. napellus . To address these research questions, we conducted a controlled experiment using potted plants to evaluate the individual and combined impacts of CTS and SA on plants exposed to Cr stress. Foliar application of CTS (0.4 g/L) or SA (0.25 mmol/L) led to significant improvements in the growth, chlorophyll content, fluorescence, and photosynthetic traits of A. napellus plants under Cr stress. The most notable effects were observed with the combined application of CTS and SA, resulting in increases in various morphological parameters, such as shoot length (2.89% and 7.02%) and root length (27.75% and 3.36%) under the Cr 1 and Cr 2 treatments, respectively. Additionally, several physiological parameters, such as chlorophyll a (762.5% and 145.56%), chlorophyll b (762.5% and 145.56%), carotenoid (17.03% and 28.57%), and anthocyanin (112.01% and 47.96%) contents, were notably improved under the Cr 1 and Cr 2 treatments, respectively. Moreover, the combined treatment of CTS and SA improved the fluorescence parameters while decreasing the levels of enzymatic antioxidants such as catalase (27.59% and 43.79%, respectively). The application also notably increased osmoprotectant parameters, such as the total protein content (54.11% and 20.07%) and the total soluble sugar content (78.17% and 49.82%) in the leaves of A. napellus in the Cr 1 and 2 treatments, respectively. In summary, these results strongly suggest that the simultaneous use of exogenous CTS and SA is an effective strategy for alleviating the detrimental effects of Cr stress on A. napellus . This integrated approach opens promising avenues for further exploration and potential implementation within agricultural production systems. Key message This explains the interactive role of chitosan and salicylic acid in alleviating chromium stress in Aconitum napellus .

Aconitum kusnezoffii Reichb. is a medicinal plant widely used in traditional Asian medicine, especially in Korea, for its potent pharmacological effects. However, its toxic alkaloids pose significant risk, making careful processing essential to reduce its toxicity. This study reviewed the plant’s processing methods, pharmacological activities, phytochemistry, toxicology, and detoxification techniques. Data from several databases, including Google scholar, PubMed, Scopus, Web science, peer-reviewed journal articles, classic herbal medicine books, and Allied and Complementary Medicine Database (AMED) were critically retrieved, organized and analyzed. The article’s findings indicate that, various ethnic groups in Asia have utilized different techniques, involving fire, water, or a combination of both to maximize the plant’s therapeutic potential and ensure safety. To date, more than 70 alkaloids, categorized into diterpenoids, norditerpenoids, and benzylisoquinolines, have been isolated from different plant’s parts (roots, leaves, stems, and flowers). These compounds exhibit various pharmacological activities including anti-inflammatory, analgesics, anti-cancer, anti-tumor, anti-arhythmic and pain-relieving properties. Despite its therapeutic potential, A. kusenzoffii . has narrow therapeutic window, meaning even small doses can be toxic. The study explored methods for reducing toxicity and detoxifying the herb emphasizing the importance of modern technologies such as propagation techniques, Systematic Evolution of Ligands by Exponential Enrichment (SELEX)-aptamer technology, and Chinmedomics in herbal medicine development. While research on this herb is extensive, gaps remain in clinical trials and efficacy studies. Further research is recommended to evaluate the quality of medicinal materials, understanding the herb’s pharmacodynamic substances, and assess long-term toxicity and clinical efficacy. Graphical Abstract

Diterpene alkaloids (DAs) are characteristic compounds in Aconitum, which are classified into four skeletal types: C18, C19, C20, and bisditerpenoid alkaloids. C20-DAs are thought to be the precursor of the other types. Their biosynthetic pathway, however, is largely unclear. Herein, we combine metabolomics and transcriptomics to unveil the methyl jasmonate (MJ) inducible biosynthesis of DAs in the sterile seedling of A. gymnandrum, the only species in the Subgenus Gymnaconitum (Stapf) Rapaics. Target metabolomics based on root and aerial portions identified 51 C19-DAs and 15 C20-DAs, with 40 inducible compounds. The highest content of C20-DA atisine was selected for further network analysis. PacBio Isoform sequencing integrated with RNA sequencing not only provided the full-length transcriptome but also their response to induction, revealing 1994 genes that exhibited up-regulated expression. Further, 38 genes involved in terpenoid biosynthesis were identified, including 7 diterpene synthases. In addition to the expected function of the four diterpene synthases, AgCPS5 was identified to be a new ent-8,13-CPP synthase in Aconitum and could also combine with AgKSL1 to form the C20-DAs precursor ent-atiserene. Combined with multiple network analyses, six CYP450 and seven 2-ODD genes predicted to be involved in the biosynthesis of atisine were also identified. This study not only sheds light on diterpene synthase evolution in Aconitum but also provides a rich dataset of full-length transcriptomes, systemic metabolomes, and gene expression profiles, setting the groundwork for further investigation of the C20-DAs biosynthesis pathway.