Abstract Ranunculaceae, commonly known as the buttercup family, represents one of the earliest‐diverging lineages among the eudicots and contains many species with horticultural and pharmaceutical importance. However, a widely accepted generic classification for this family remains lacking. In this study, we present, to our knowledge, the most comprehensive phylogenetic analysis of Ranunculaceae (227 species), representing almost all currently recognized genera and generic synonyms to date based on five plastid and nuclear loci. We reconstructed the phylogenetic relationships within Ranunculaceae using likelihood, parsimony, and Bayesian approaches. We used the SOWH test to assess the alternative topologies. Our results indicate that Glaucidioideae is the earliest‐diverging lineage in Ranunculaceae, followed by Hydrastidoideae, and Coptidoideae and Thalictroideae are successive sisters to Ranunculoideae. The SOWH test rejects Glaucidioideae as sister to Hydrastidoideae. The sister relationship between Thalictroideae and Adonideae of Ranunculoideae is also rejected. Within Ranunculoideae, the monophyly of each of all 10 tribes is strongly supported. The majority of genera of Ranunculaceae are supported as monophyletic except for Anemonastrum , Enemion , and Oxygraphis . The monophyly of each of these three genera is also rejected by the SOWH tests. Paroxygraphis was included in a molecular phylogenetic analysis for the first time and was nested in Oxygraphis . Based on the results of our molecular data analyses, as well as the results of previous studies, a new subfamilial, tribal, and generic classification of Ranunculaceae is proposed, in which 5 subfamilies, 14 tribes (including one newly described tribe), and 58 genera (including three newly described genera) are recognized.

Chloroplast genomes provide conserved yet informative sequences useful for inferring plant evolution and species identification. Genus Aconitum consists of around 300 traditional Indian and Chinese medicinal plant species, many native to mountainous regions, and known to be highly poisonous due to toxic diterpene alkaloids. Their accurate identification and classification are vital for traditional medicine systems, particularly for ensuring their safe use. A consistent quadripartite structure was identified across all chloroplast genomes, comprising the typical large single copy (LSC), small single copy (SSC), and two inverted repeats (IR) regions. Pan-plastome analysis unveiled 72 core and nine accessory genes, indicating an open pan-plastome characteristic. In-depth nucleotide-level homology analysis revealed that homologous genes of all accessory genes are present in all other genomes, implying the need for better chloroplast genome annotation tools that can identify all putative genes from such conserved genomes. Notably, the order of all core and accessory genes remained highly conserved across all analysed genomes, underscoring the overall evolutionary stability despite the diversity of accessory genes. Members of some core pathways are relatively absent on the chloroplast genome, suggesting their potential presence on the nuclear genome, which will be revealed after their nuclear genome sequencing. Our phylogenetic results largely supported the morphological classification, with distinct Lycoctonum and Aconitum subgenera clustering, further validating the gross accuracy except for A. flavum, suggesting a putative morphological classification discrepancy or inaccurate classification. This comparative analysis reveals a highly conserved chloroplast genome architecture across Aconitum while documenting measurable plastome-level variation among 73 species. The chloroplast phylogeny highlights instances of non-monophyletic clustering among conspecific accessions and unexpected placement of certain taxa, indicating complex evolutionary histories within the genus. Such patterns may reflect a combination of processes, including incomplete lineage sorting, chloroplast capture, or sample misidentification, rather than definitive phylogenetic incongruence. While chloroplast genomes alone cannot fully resolve evolutionary relationships in a group shaped by hybridization and polyploidy, the results provide valuable insights into plastome evolution and identify key lineages and taxa that warrant further investigation using nuclear genomic and integrative approaches.

A comprehensive review of the traditional usages, phytochemistry, pharmacology, toxicology, quality control and other applications of Aconitum kusnezoffii Reichb.

Anemone sylvestris Linnaeus (1753) is a species in the Ranunculaceae family within the genus Anemone. It is mainly distributed in China, Croatia, and Poland. The chloroplast genome has a total length of 161,033 bp, comprising a large single-copy region of 81,144 bp, a small single-copy region of 17,393 bp, and two inverted repeat regions of 31,248 bp each. The GC content is 37.54%. A total of 134 genes were annotated, including 90 protein-coding genes, 8 rRNA genes, and 36 tRNA genes. Phylogenetic analysis using the maximum likelihood method robustly supported the close relationship of A. sylvestris with other Anemone species, namely A. raddeana, A. reflexa, and A. altaica, forming a well-defined monophyletic clade. The complete chloroplast genome of A. sylvestris reported here provides a useful resource for subsequent phylogenetic, identification, and resource related studies.

Black cumin ( Nigella sativa L.) is a spice crop from the Ranunculaceae family and is produced extensively across the Mediterranean region, Asia Minor, Northern Africa, and Southern Europe. It has significant nutritional and pharmaceutical values, and global popularity and demand are increasing due to its widespread use in traditional and modern medicine in diverse forms. In Ethiopia, black is the largest cash crop, ideal for its medicinal properties and economic benefits. Ethiopians continue to produce and consume a relatively small amount of black cumin, despite its widespread use for medicinal purposes and its cultural significance. This is due to a lack of improved technology, insufficient germplasm, and improved varieties, limiting research extension. The present review highlights the current status of black cumin cultivation, its therapeutic potential, and its traditional use in Ethiopia, as well as the challenges to its production and commercialization. The text emphasizes the significance of genetic resource conservation, sustainable utilization, and the need for further research to improve production and commercialization. Addressing these issues will allow black cumin to reach its full potential, contributing to food security and economic development in Ethiopia. Therefore, this review aims to assess and document the status, use, conservation, and sustainable utilization of black cumin, as well as their gaps and prospects.

Aconitum carmichaelii Debeaux is a perennial herbaceous plant within the Ranunculaceae family. The processed lateral roots of A. carmichaelii, known as "Fuzi", are widely used as traditional Chinese medicinal materials for the treatment of various human diseases such as rheumatoid arthritis, cardiovascular diseases, and tumors in Asian countries (He et al., 2023). A. carmichaelii has been cultivated for over 1,000 years (Yu et al., 2016). Jiangyou, in Sichuan, China, has been considered the geo-authentic cultivating area of A. carmichaelii (Zhou et al., 2015). An outbreak of soft rot in A. carmichaelii caused up to 60%-80% of plant death in Jiangyou in April, 2024. Soft rot blackened lesions may develop on infected stems, roots, or both, leading to the collapse and wilting of the leaves, and death of the plant. To isolate the causal agent, tissue fragments (5 mm × 5 mm) from lesion margins of 5 symptomatic plants were surface-sterilized by soaking in 75% ethanol for 30 s, and 2% NaOCl for 1 min, followed by rinsing with sterilized water for 5 times. The materials were then macerated in sterile water, and streaked onto Luria-Bertani (LB) agar plates. After incubation at 28°C for 48 h, four bacterial isolates (SR-1, SR-2, SR-3, and SR-5) were picked and purified on LB plates. Single colonies of these isolates were milky white, round, opaque, glossy, and smooth-edged, with 0.2-0.3 cm diameter. All the strains were Gram-negative. Genomic DNA was extracted from each strain using Bacterial DNA Kit (Sangon, Shanghai) and was used as template for the amplification of 16S rRNA, atpD, gyrA and recA genes (Menna et al., 2009; Wang et al., 2017). Sequences of strains SR-1, SR-2, SR-3, and SR-5 have been deposited in the GenBank under accession numbers PV776673-PV776676 (16S rRNA), PV799309-PV799312 (atpD), PV799313-PV799316 (gyrA), and PV799317-PV799320 (recA). Phylogenetic tress based on 16S rRNA gene and multilocus sequence analysis (MLSA) of concatenated 16S rRNA, atpD, gyrA, and recA sequences were constructed using the Neighbor-Joining method in MEGA X with 1,000 bootstrap replicates. SR-1, SR-2, SR-3, and SR-5 clustered together with Pectobacterium versatile CFBP 6051T (MK226529) with similarities ranging from 97.66% to 98.00% in the phylogenetic tree based on 16S rRNA gene. Consistently, MLSA-based phylogeny revealed that SR-1, SR-2, SR-3, and SR-5 clustered with P. versatile CFBP 6051T (RMBU01000000), sharing 98.73%, 98.70%, 98.70%, and 98.70% identities, respectively. To verify the pathogenicity of the strains, bacterial suspension of each isolate (1×10⁸ CFU/mL, 1 mL) was injected to the roots of A. carmichaelii plants using a sterilized syringe (5 plants per treatment, 3 biological replicates on cultivar MianFu No. 1). Sterilized water was used as a negative control. The control plants were healthy throughout the experiment. SR-1, SR-2, and SR-5 induced wilting of the plants at 3 days post inoculation (dpi), while SR-3 did at 5 dpi. Plants inoculated with test strains all died at 10 dpi. Strains were successfully reisolated from the symptomatic plants and confirmed as P. versatile by the methods mentioned above. P. versatile has been reported as causing pathogen of potato, cabbage, tobacco, and Hydrangea macrophylla (Chen et al., 2022; Handique et al., 2023; Lu et al., 2023; Su et al., 2023). To our knowledge, this is the first report of P. versatile causing soft rot in A. carmichaelii in China, expanding its known host range. Soft rot posed remarkable threat to the yield and quality of A. carmichaelii. Further researches on epidemiology and management are warranted.

‌Medicinal plant black cumin (Nigella sativa L.) is an annual herb from the Ranunculaceae family, which has been used us food, medicine, and therapeutic purposes for centuries. There are 20 to 24 species of black cumin in the world, and it is reported that 12 to 15 of these species are found in the flora of various countries. Black cumin is one of the valuable medicinal and aromatic plants that can be easily cultivated in the climatic conditions of Baghlan province. This research was conducted in completely randomized design (CRD) at the research farm of Agriculture Faculty, Baghlan University in 2024. In this study, four levels of phosphorus (0, 65, 130, and 195 kg/ha) were applied. The main objective of this experiment was to examine the morphological traits and yield of the medicinal plant black cumin under different levels of phosphorus. The results indicate that phosphorus fertilizer had a significant effect on the morphological traits of the medicinal plant black cumin. Phosphorus fertilizer at doses of 130and 195 kg/ha had a positive and significant impact on plant height, the number of capsules per plant, the number of seeds per capsule, and the yield of this plant. These results highlight the importance of adjusting appropriate phosphorus levels to improve the vegetative growth and yield of the medicinal plant black cumin.

Over the past two decades, the global use of medicinal plants as complementary and adjuvant therapies has increased significantly for the management of various diseases. Nigella sativa, commonly known as kalonji, black cumin, or Roman coriander, is among the most extensively studied evidence-based medicinal herbs. It belongs to the family Ranunculaceae and is an annual flowering plant characterized by bluish-white flowers and small, black, flattened, oblong seeds resembling onion seeds. Synonyms include Nigella indica and Nigella truncata, and the plant is widely cultivated in India, the Middle East, and Mediterranean regions, thriving in both cold dry and warm humid climates. The seeds, fruits, and seed oil of N. sativa are extensively used in both culinary and medicinal applications. Numerous pharmacological properties have been documented, including antihypertensive, diuretic, antidiarrheal, analgesic, antimicrobial, immunomodulatory, antioxidant, hepatoprotective, and gastroprotective effects. In the United States, N. sativa is classified as Generally Recognized as Safe (GRAS) by the Food and Drug Administration for consumption. Phytochemical analyses reveal the presence of fixed oils, proteins, alkaloids, saponins, and essential oils, with thymoquinone identified as the principal bioactive compound responsible for many therapeutic actions. This review aims to highlight the medicinal significance of Nigella sativa, promote greater scientific interest in herbal therapeutics, and encourage further exploration of its potential in disease prevention and treatment.

Abstract The genus Helleborus belongs to the family Ranunculaceae and comprises about 22 species, which are distributed across Europe and West Asia. Hellebores are popular as garden perennials, pot plants, cut flowers, and medicinal plants. The present study aimed to evaluate the effects of sucrose level (20, 30, 40, and 50 g·L −1 ), mineral salts content (50%, 100%, 150% of Murashige and Skoog 1962 medium – MS), growth regulators (BAP – 6-benzylaminopurine, 2iP – 6-(γ,γ-dimethylallylamino) purine, kinetin, GA 3 – gibberellic acid, IBA – indole-3-butyric-acid), and white light sources (fluorescent and LED) for multiplication and in vitro rooting Helleborus of ‘Molly’s White’ and ‘Anna’s Red’. Additionally, the post-effect of white, blue, and red light (LED) used during the storage of shoots at a temperature of 15°C for 3 months in glass jars and plastic vessels on the subsequent in vitro rooting and on ex vitro acclimatization was investigated. The results presented here showed that sucrose and nitrogen salts were more critical for axillary shoot branching and subsequent in vitro rooting in both hellebore genotypes than the concentration of growth regulators in the multiplication and rooting media. Low level of sucrose (20 g·L −1 ) and standard content of mineral salts (100% in the MS medium) were optimal for the development of axillary shoots. The medium composition used in the multiplication experiments markedly affected the rooting ability of both cultivars. The sucrose/nitrogen salts ratio in the multiplication medium was more critical for subsequent rooting than the type and concentration of growth regulators or the composition of the rooting media. Genotype ‘Molly’s White’ exhibited the best rooting after multiplication on the medium with high (40 g·L −1 ) and low (20 g·L −1 ) sucrose levels, but ‘Anna’s Red’ only on the medium with low sucrose content (20 g·L −1 ). The white LED increased the multiplication and rooting rates of both hellebores compared with white fluorescent light. Plastic vessels significantly increased root number in both hellebore genotypes. The post-effect of white, blue, and red light (LED) used during the storage of cultures had a weak influence on ex vitro acclimatization compared to in vitro rooting. Genotypic differences in multiplication rate, rooting ability, and ex vitro acclimatization efficiency were observed.

In this study, new distribution information is given for a total of 20 taxa, 11 of which are endemic, including Beta macrorhiza and Akhania canescens belonging to the Amaranthaceae family, Eupatorium cannabinum, Lactuca intricata and Rhaponticum altaicum belonging to the Asteraceae family, Leontice ewersmannii belonging to the Berberidaceae family, Onosma intertexta belonging to the Boraginaceae family, Velezia hispida, Gypsophila osmangaziensis, G. tubulosa, G. turcica, Silene aeoniopsis, S. caryophylloides subsp. masmenaea and S. nerimaniae belonging to the Caryophyllaceae family, Hyssopus officinalis, Marrubium sivasense, Sideritis amasiaca and S. gulendamii belonging to the Lamiaceae family, Ranunculus peltatus subsp. fucoides belonging to the Ranunculaceae family, and Alchemilla caucasica belonging to the Rosaceae family. In addition, IUCN threat categories are proposed for some of these taxa. New population discoveries are significant in contributing to the chorology and conservation of the species.

Clematis L., a significant genus of climbing plants within the Ranunculaceae family, boasts widespread germplasm resources distributed across temperate to tropical regions globally, with Asia preserving particularly abundant native populations. This review systematically summarizes recent advances in Clematis research: in terms of physiological characteristics, the research focuses on the evolution of plant classification, chromosomal evolutionary features revealed by karyotype analysis, and studies on genetic diversity and phylogenetic relationships based on molecular markers; in breeding methods, it summarizes the two major technical systems of sexual and asexual reproduction; regarding ornamental traits, it emphasizes the molecular mechanisms of flower color and form development, and synthesizes breakthroughs in techniques for flowering period regulation and research on the biosynthesis pathways of floral scent metabolites; in the field of stress resistance mechanisms, it thoroughly examines physiological responses and molecular adaptation mechanisms under abiotic stresses such as UV radiation, drought, high temperature, and intense light, and outlines research progress on pathogen types of major diseases; in studies of medicinal value, it highlights the material basis and mechanisms of pharmacological activities including anti-inflammatory, analgesic, and antitumor effects. Through multidimensional comprehensive analysis, this review aims to elucidate the comprehensive development potential of Clematis, providing theoretical foundations and practical guidance for germplasm resource innovation, breeding of high-ornamental-value cultivars, and stress resistance applications.

Ranunculus ternatus Thunb. (RT), a species in the Ranunculaceae family, has been officially recognized in traditional Chinese medicine and included in the Chinese Pharmacopoeia since 1977. Its dried root has been widely used in Chinese medicine to treat a variety of ailments, including stomatitis, tuberculosis, and other inflammatory conditions. More recent research has expanded its potential, showing promise in the treatment of modern diseases such as diabetes, gastric carcinoma, and ovarian cancer. It is widely cultivated in several Chinese provinces. In February 2025, leaf spot disease was observed in a 2-ha field in Funan County, Anhui Province (115°16'32''E, 32°24'25''N), with a disease incidence of 30%. Early symptoms included brown spots or streaks on the leaves. As the disease progressed, large, irregular brown spots merged, eventually covering the entire leaf and causing leaf death. A layer of grey mold was observed on affected leaves. Leaf tissue segments (1 cm²) were surface-sterilized with 75% ethanol for 30 s, followed by 5% NaClO for 1 min, rinsed three times with sterile water, and cultured on potato dextrose agar (PDA). After mycelial growth, hyphal tips were transferred to fresh PDA plates, purified through subculturing, and preserved. Five pathogenic strains with similar morphological features were isolated, and strain fy11-5 was selected for further characterization. After growing on PDA at 25 °C for 6 days, the colony produced white, fluffy mycelia that turned grey-brown. Microscopic examination revealed ellipsoidal to spherical, hyaline conidia (9.65–14.15 μm × 4.93–5.74 μm). Conidiophores were clustered, greyish, and branched, with subglobose conidiogenous cells, resembling a grape-like structure. Based on these features, the isolate was identified as Botrytis cinerea (Ellis 1971). Pathogenicity testing was conducted by inoculating R. ternatus Thunb. leaves with a 5 × 5 mm mycelial plug from an actively growing colony; control leaves received a sterile PDA plug. Disease progression was assessed at 3, 5, and 10 days post-inoculation in a controlled environment at 25 °C, 75% relative humidity, under a 12-h light/12-h dark cycle. Grey lesions developed on inoculated leaves, leading to wilting, while control leaves remained asymptomatic. The fungus was re-isolated from symptomatic tissue, and its morphology matched the original isolate, fulfilling Koch’s postulates. Genomic DNA of strain fy11-5 was PCR-amplified for the ITS, HSP60, RPB2, and G3PDH genes, yielding sequences of 519 bp, 1117 bp, 882 bp, and 1018 bp, respectively. The primers used were ITS1/4 (White et al. 1990), HSP60-F/HSP60-R, RPB2-F/RPB2-R, and G3PDH-F/G3PDH-R (Staats et al. 2005). BLAST analysis of the NCBI GenBank showed 99-100% identity with B. cinerea strains (GenBank accession nos. PV776292, PV791144, PV791145, PV791146 for ITS, G3PDH, HSP60, and RPB2, respectively). Phylogenetic analysis of the combined sequences confirmed that fy11-5 is B. cinerea. Thus, both morphological and molecular evidence identified strain fy11-5 as B. cinerea, responsible for leaf spot disease on R. ternatus Thunb. This is the first report of B. cinerea causing leaf spots on R. ternatus in Anhui Province, China, providing a basis for controlling the disease and its economic impact.

Berberine is a bioactive compound found in the rhizomes, stems, and fruits of plants belonging to the Berberidaceae, Ranunculaceae, Menispermaceae, Papaveraceae, and Rutaceae families. Although berberine is widely distributed throughout all parts of plants, the bark and root contain significantly higher concentrations of berberine compared with other plant parts. Berberine possesses various pharmacological properties, including antitumor, antibiotic, analgesic, antihyperlipidemic, and anti-inflammatory effects. The bioactive components of berberine are considered promising for the prevention and treatment of various diseases. Berberine has been shown to exert beneficial effects in the management of obesity, diabetes mellitus, cardiovascular and gastrointestinal diseases, cancer, and viral infections. It enhances insulin sensitivity, inhibits gluconeogenesis in the liver, and regulates gut microbiota. While some side effects associated with berberine use have been reported, its overall use is generally regarded as safe, with potential beneficial effects in the treatment of specific diseases. However, it is not appropriate to generalize its beneficial effects in disease prevention. This review highlights the chemical properties, dietary sources, absorption mechanisms, metabolism, biosynthesis, and the promising health benefits of berberine. Current evidence and discussions are examined, and several suggestions for future research are proposed.

Nigella sativa L. (black cumin) is an annual herb of the Ranunculaceae family, traditionally used in Middle Eastern and Asian medicine for the management of various health conditions. Its seeds and oils are rich in bioactive compounds, including alkaloids, essential fatty acids, nigellone, saponins, and thymoquinone (TQ), which contribute to diverse pharmacological activities such as anticancer, antidiabetic, anti-inflammatory, antimicrobial, antioxidant and hepatoprotective effects. In the present study, oils obtained from Turkish and Syrian N. sativa seeds through cold pressing were investigated. Phytochemical composition was characterized using Gas Chromatography (GC) and High Performance Liquid Chromatography (HPLC), with particular focus on fatty acid profiles and TQ content. Additionally, tyrosinase inhibitory activities of the fixed oils and TQ were evaluated. It was noted that variations in the composition of seed samples originating from different countries influence the phytochemical features of the plant materials. Accordingly, maintaining genetic diversity and implementing appropriate cultivation strategies are essential for the sustainability of agricultural production.

Background: Aconitum heterophyllum (Family Ranunculaceae) is a perennial herb native to the western Himalayas (2,500-4,000 m) and is used in Ayurvedic and Unani systems for a variety of disorders including diarrhoea, fevers, indigestion, respiratory ailments and nervous-system conditions.Its therapeutic value is underpinned by its tuberous roots, which contain a gamut of bioactive diterpene alkaloids.Objective: This work aims to synthesize available information on the botany, phytochemistry, pharmacology, traditional uses, propagation/cultivation and conservation status of A. heterophyllum, highlighting its medicinal promise and the urgency of sustainable use. Methods:A comprehensive literature survey was conducted, drawing on botanical monographs, traditional medicinal texts, and modern phytochemical/pharmacological research.Key data on morphology (roots, stem, leaves, flowers, seeds), habitat/distribution, active constituents (atisine, heteratisine, hetisine and related alkaloids), pharmacological activities (anti-diarrheal, antipyretic, anti-inflammatory, immune-modulatory) and conservation/propagation strategies were extracted and analyzed. Results:The species thrives in moist, humus-rich soils in temperate to alpine slopes of the Himalayas and has distinct tuberous roots which serve as the principal medicinal part.Phytochemical studies reveal numerous alkaloids (e.g., Atisine) that are comparatively less toxic than those in other Aconitum species, supporting its relatively safer medicinal profile.Pharmacological investigations confirm antisecretory, antimotility, anti-inflammatory and hepatoprotective effects.However, wild populations face serious threats from over-harvesting, habitat degradation and climate-driven range contraction, while propagation efforts (via seed and vegetative methods) remain limited but vital. Conclusion:Aconitum heterophyllum represents a valuable medicinal herb with strong ethnopharmacological grounding and encouraging modern scientific support.Nonetheless, translating its traditional promise into modern therapeutic application demands further work particularly standardization of active compounds, rigorous clinical evaluation, scalable cultivation/propagation, and robust conservation strategies to preserve natural populations.

BackgroundPulsatilla species, which belong to the Ranunculaceae family, have garnered significant attention due to their remarkable medicinal attributes and ornamental value. In the present study, four mitochondrial genomes (mitogenomes) of Pulsatilla species were assembled and analyzed. The aim was to lay a research foundation for unraveling the genetic interrelationships among these species and the identification of Traditional Chinese Medicine from Pulsatilla species.ResultsThe mitogenomes of P. chinensis, P. chinensis var. kissii, P. cernua, and P. dahurica were assembled into single circular DNA molecules, with lengths of 878,988 bp, 684,203 bp, 747,621 bp, and 824,625 bp, encoding 53, 40, 40, and 49 protein-coding genes, 13, 14, 20, and 33 transfer RNA genes, and 3, 3, 4, and 3 ribosomal RNA genes, respectively. Repeat sequence analysis found a large number of simple sequence repeats (SSRs) and long repeat sequences (LRSs) in four mitogenomes. P. chinensis, P. chinensis var. kissii, P. cernua and P. dahurica had 43, 4, 6, 3 LRSs longer than 1 kb, respectively. Codon bias and RNA editing sites in the protein-coding genes of the four mitogenomes were similar. Furthermore, numerous collinear sequences were identified among the four mitogenomes, and homologous fragments were found by comparing them with their plastome sequences. Based on the phylogenetic trees of mitochondrial PCGs, P. chinensis, P. chinensis var. kissii, P. cernua, and P. dahurica clustered into a common subclade.ConclusionsThe comprehensive analysis of the four Pulsatilla mitogenomes revealed that the genome size, gene composition, and distribution of repeat sequences display variability. This finding offers novel perspectives into the evolution of related species, thereby enriching our understanding of their genetic underpinnings and potential for further exploration in diverse fields.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-025-07527-1.

ABSTRACTThe Ranunculaceae family, encompassing approximately 2500 species across 50 genera, includes several taxa of medicinal importance. This study presents de novo assembled chloroplast (cp) genomes for three species: Aconitum violaceum Jacquem. ex Stapf, Caltha palustris L., and Delphinium denudatum Wall. ex Hook.f. & Thomson. All three cp genomes exhibited the typical angiosperm quadripartite structure—comprising a large single‐copy region, a small single‐copy region, and a pair of inverted repeats (IRa and IRb)—with total lengths of 154,523 bp (A. violaceum), 155,057 bp (C. palustris), and 154,228 bp (D. denudatum). Genome annotation identified 111–112 unique genes, including 77–78 protein‐coding genes, 30 tRNAs, and four rRNAs. Notably, rps16 and rpl32 were absent in A. violaceum and D. denudatum, whereas infA was missing in C. palustris. Comparative analysis revealed high synteny, with no major genomic rearrangements, although minor IR boundary shifts were observed involving rps19, ycf1, and ndhF. Codon usage showed a pronounced bias toward A/T‐ending codons (RSCU > 1.0), with leucine and isoleucine being the most frequently encoded amino acids. Simple sequence repeat (SSR) analysis detected 65–93 SSRs per genome, predominantly A/T‐rich mononucleotide motifs. Maximum likelihood phylogenetic analysis of 76 complete cp genomes confirmed the monophyly of Aconitum L., Caltha L., and Delphinium Tourn. ex L., placing D. denudatum within Delphinium and grouping A. violaceum with Aconitum tanguticum in subgenus Aconitum. These findings provide genomic resources to support improved phylogenetic resolution, molecular evolution studies, conservation genetics, and medicinal plant research within Ranunculaceae.

ABSTRACTBenzoxazinoids are secondary metabolites produced in monocotyledons and some dicotyledons from the Acanthaceae, Ranunculaceae, Scrophulariaceae, Plantaginaceae, and Lamiaceae families. Benzoxazinoids are commonly consumed in bread and cereal products that have been ingested for decades, which supports the safe use of benzoxazinoids as food ingredients. Powdered corn leaf extract (UP165), trade name Maizinol, containing 0.2%–0.3% w/w benzoxazinoid 6‐methoxy‐2‐benzoxazolinone (6‐MBOA), was the subject of genetic toxicity studies conducted according to Organisation for Economic Co‐operation and Development (OECD) Test Guidelines (TG). The toxicity tests evaluated the safety of corn leaf extract (as Maizinol containing 0.2%–0.3% 6‐MBOA) using a bacterial reverse mutation assay (OECD Test Guideline 471) at doses of 313, 625, 1250, 2500, or 5000 μg/plate; in vitro mammalian chromosomal aberration test (OECD TG 473) at concentrations of 9.77, 19.5, 39.1, or 78.1 μg/mL; and in vivo mammalian erythrocyte micronucleus test (OECD TG 474) at a dose of 750, 1500, or 3000 mg/kg body weight/day. Results demonstrate that Maizinol (UP165), at maximum test concentrations of 5000 μg/plate in the bacterial mutagenicity study, 78.1 μg/mL in the mammalian chromosomal aberration test, and a dose of 3000 mg/kg body weight in the rat micronucleus test, did not elicit genotoxic or mutagenic effects under the reported test conditions.

ABSTRACTMaize (Zea mays) has been consumed by humans for millennia and represents the third most abundant crop grown globally. Maize and maize‐derived products have a long history of safe consumption from bread and other cereal products in human diets worldwide. Aside from key dietary components like carbohydrates and proteins, the corn plant contains endogenous biomolecules like benzoxazinoids. Benzoxazinoids are a group of secondary metabolites produced in monocotyledons and some species of dicotyledons from the Acanthaceae, Ranunculaceae, Scrophulariaceae, Plantaginaceae, and Lamiaceae families, and benzoxazinoids play a vital role in plant physiology. The current research aims to evaluate the safety of Maizinol (UP165), an ethanolic corn leaf extract containing 0.2%–0.3% of the benzoxazinoid, benzoxazolinone 6‐methoxy‐2‐benzoxazolinone (6‐MBOA) in Sprague Dawley rats. Animals were orally dosed with 0‐, 750‐, 1500‐, or 3000‐mg Maizinol/kg body weight/day for 90 days and observed for mortality and morbidity. Results revealed that Maizinol was well tolerated at all tested doses, as no significant effects were observed in hematological, biochemical, and histological endpoints at all doses. A subchronic 90‐day toxicity no‐observed‐adverse‐effect level for UP165 in Sprague Dawley rats was determined to be 3000 mg/kg body weight/day.

A multi-level strategy based on comprehensive two-dimensional liquid chromatography-Q-Orbitrap-mass spectrometry combined with PLS regression model and RT-Ensemble Pred model to intelligently distinguish different geographical locations of Huanglian sample.