Long-term organogenic callus cultivation of Ranunculus illyricus L.: a blueprint for sustainable ex situ conservation of the species in urban greenery.

The growing trend of introducing wild plant species into urban environments necessitates the identification of novel species adapted to prevailing conditions. A promising reservoir of such species may be xerothermic communities where Ranunculus illyricus occurs. This study aimed to establish a micropropagation protocol for R. illyricus using indirect organogenesis. The protocol includes initiation of culture from various explants, callus proliferation, shoot regeneration, multiplication, and concurrent rooting. Callus appeared on most types of vegetative explants tested, but stolons were considered the best due to their good availability, high disinfection (85%), and robust callus production (maximum increase - 363.1%). The growth rate of the callus fresh matter (CFM) obtained from stolons was calculated. Greater CFM was obtained on the medium with the supplemented picloram 8.0 mg L- 1 with kinetin 5.0 mg L- 1 and in second part of experiment on medium with the addition of 2,4-D (2,4-dichlorophenoxyacetic acid) 2.0 mg L- 1 alone or picloram 6.0 mg L- 1 with kinetin 8.0 mg L- 1. Shoot organogenesis was observed on macronutrients B5 (Gamborg medium), micronutrients MS (Murashige and Skoog) medium with the addition of 2.0 mg L- 1 IBA (indole-3-butyric acid) and 4.0 mg L- 1 BAP (6-benzylaminopurine). To document the process of callus differentiation, microscopic preparations were prepared. Subsequently, the regenerated plants underwent acclimatisation and their growth in an ex situ collection was monitored over three growing seasons. In particular, in vitro-origin plants exhibited developmental patterns similar to those of their seed-origin counterparts. The incorporation of R. illyricus into urban landscapes not only increases aesthetic appeal, but also ensures the preservation of valuable genetic resources for this rare species, potentially contributing to effective ex situ conservation in the future. This marks the first scientific report on in vitro cultures of R. illyricus.

Ranunculin, Protoanemonin, and Anemonin: Pharmacological and Chemical Perspectives.

Ranunculin, a glucoside, serves as a chemotaxonomic marker in Ranunculaceae plants. When these plants are damaged, an enzyme ß-glucosidase triggers the conversion of ranunculin into protoanemonin through hydrolysis. Subsequently, protoanemonin undergoes cyclodimerization to form anemonin. The inherent instability of ranunculin and the rapid dimerization of protoanemonin render them unsuitable for use in biological assays. Conversely, anemonin stands out as the optimal molecule for bioassays and demonstrates diverse biological properties, including anti-inflammatory, anti-infective, and anti-oxidant effects. Among these, anemonin exhibits the greatest promise in addressing conditions such as arthritis, cerebral ischemia, and ulcerative colitis. Its potential medical uses are enhanced by its capacity to inhibit nitric oxide synthesis and successfully counteract lipopolysaccharide-induced inflammation. This review describes the chemistry and biological properties of anemonin and its precursors, including discussions on extraction, isolation, synthesis, and investigations into bioactivity and pharmacokinetics.

The complete chloroplast genome of Pulsatilla chinensis f. alba D. K. Zang (Ranunculaceae, Pulsatilla Miller).

Pulsatilla chinensis f. alba D. K. Zang 1993 is a forma of Pulsatilla chinensis (Bge.) Regel, the root of P. chinensis is traditional Chinese medicine called Pulsatillae radix. The biggest difference between P. chinensis f. alba and P. chinensis is that P. chinensis f. alba sepals is white. The complete chloroplast genome of P. chinensis f. alba was sequenced using the Illumina NovaSeq platform for the first time. The lengths of the genome, large single-copy (LSC), small single-copy (SSC), two inverted repeats (IRs), and GC content were 163,654 bp, 82,355 bp, 19,069 bp, 31,115 bp, and 37.2%, respectively. It had 134 genes, including 90 protein-coding genes, 36 tRNA genes, and eight rRNA genes. The maximum-likelihood tree indicated that P. chinensis f. alba had a closer relationship with P. chinensis. This study would provide a theoretical basis for the further study of Pulsatilla plants genetics phylogenetic research.

Micromorphological and phytochemical profiling of Delphinium suave Huth. from Hindukush range, Lower Dir Khyber Pakhtunkhwa Pakistan.

This is a very first attempt to study various parameters of a medicinal plant, Delphinium suave Huth. The plant is erect, geophytic, herbaceous, with tuberous root, trifid in a palmatipartite, strigose cuneate leaf and white spurred zygomorphic flower. The root was isodiametric phellem with single non-glandular trichomes. The stem revealed single-layered cuticle, multiseriate epidermis, cortex, pith ray and uniserate bowed non-glandular trichomes. The leaf was amphistomatic, showed tapering trichomes, prismatic crystals and ranunculaceous stomata with circumference 144.66-182.67 µm. Pollen grains in Light Microscopy (LM), were prolate, spheroidal trizonocolpate, isopolar, radiosymmetric, scabrate, elliptic and monads. Scanning Electron Microscope (SEM) pollen surface was scabrate, monad, size varied from 18.06 to 16.67 µm, colpus to inaperturate, tricolpate, ornamented, echinus, isopolar, isodiametric and circular. SEM roots showed sclerenchymatic tissues, stellate, glandular, non-glandular trichomes and crystals. The stem showed scalariform, pitted vessels, warty protuberances, unicellular, silicified, non-glandular trichomes. Leaves powder revealed, simple, unicellular, tapered headed, uniseriate, sessile, capitate, unbranched glandular, non-glandular, trichomes with crystals. Capitate, stellate, circular, unicellular, branchy trichomes were observed for the first time through SEM. Powder drug study of root, stem leaves through LM revealed different tissues. Preliminary phytochemical revealed alkaloids, anthocyanins, anthraquinones, coumarins, flavones, mucilages, saponins, steroids, terpenoids, volatile oils and proteins. GC/MS showed 36 compounds in roots, 33 in stem while 40 in leaves. Fluorescence analysis of roots, stem and leaves showed variations in color when treated with chemicals. This study will assist pharmacognostic exploration, authentication from adulterants/allied species for consistent quality, resulting in safe use, preservation and efficacy. RESEARCH HIGHLIGHTS: This was first attempt on pharmacognostic study on D. suave Huth. which could be used as a foundation for identifying and authenticating the specie from other allied species by these morphological, anatomical, GC/MS profiling, phytochemical analysis and fluorescence analysis.

Investigations on the morpho-anatomy and histochemistry of Aconitum napellus L.

Aconitum napellus L. is a popular medicinal plant extensively used in homeopathy. This article provides detailed morphology and microscopy, including the anatomical and histochemical features of the herb, to aid authentication and quality control. In cross-section, the root in secondary growth shows the phloem surrounded by pericyclic fibers and a well-developed xylem. The stem is irregular in outline, displaying unicellular trichomes and many free collateral vascular bundles encircling the pith. The leaf is dorsiventral, hypostomatic with anomocytic and anisocytic stomata, and shows non-glandular trichomes. The floral parts are characterized by uniseriate epidermises, homogeneous mesophyll, anomocytic stomata on the abaxial surface, trichomes, and oval pollen grains. The tissue fragments in powdered herbs show these characteristics and have numerous starch grains with thimble-shaped, linear or star-shaped hilum. The detailed macroscopic and microscopic analysis provided in this study can help in the authentication and quality control of A. napellus raw materials. RESEARCH HIGHLIGHTS: Key anatomical, micromorphological, and microchemical features of Aconitum napellus are described. The results of the study can support the taxonomy of the genus Aconitum. Morphological standardization of the species reported here is helpful in the quality control of this herb.

A new C19-diterpenoid alkaloid in Aconitum georgei Comber.

Nine diterpenoid alkaloids were isolated from Aconitum georgei Comber belonging to the genus Aconitum in Ranunculaceae family. Their structures were determinated by using HR-ESI-MS and 1 D/2D NMR spectra as geordine (1), yunaconitine (2), chasmanine (3), crassicauline A (4), forestine (5), pseudaconine (6), 14-acetylalatisamine (7), austroconitine B (8), and talatisamine (9). Among them, compound 1 is a previously undescribed aconitine-type C19-diterpenoid alkaloid, and compounds 3, and 5-9 have not previously been isolated from this species. The results of in vitro experiments indicated that new compound 1 possesses mild anti-inflammatory activity, which inhibited the production of NO in LPS-activated RAW 264.7 cells with an inhibition ratio of 29.75% at 50 µM.

Complete chloroplast genome of Adonis pseudoamurensis W.T.Wang (Ranunculaceae).

Adonis pseudoamurensis W.T. Wang 1980 is an important traditional medicinal plant used for the treatment of cardiac diseases. The complete chloroplast (cp) genome of Adonis pseudoamurensis is reported for the first time in this study. The circular cp genome is 156,917 bp in length, consisting of a large single-copy region (86,262 bp), a small single-copy region (18,067 bp), and two inverted repeat regions (26,294 bp). The genome encodes 129 genes, comprising 84 protein-coding genes, 37 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. Phylogenetic analysis showed that A. pseudoamurensis is closely related to A. amurensis.

Complete chloroplast genome data of Ranunculus membranaceus (Ranunculaceae), an important medicinal plant species.

The perennial alpine herb Ranunculus membranaceus (Ranunculaceae) has significant medicinal value. The complete chloroplast genome of R. membranaceus was sequenced by high-throughput Illumina sequencing Platform Illumina NovaSeq 6000. The circular genome is 156,028 bp in size, including two inverted repeats (IRs) of 25,361 bp, a large single-copy (LSC) region of 85,491 bp, and a small single-copy (SSC) region of 19,815 bp. A total of 128 genes were annotated, namely 84 protein-coding genes (PCGs), 36 tRNA genes, and eight rRNA genes. Two phylogenetic trees of 18 species of the tribe Ranunculeae species were constructed with Meconopsis punicea as the outgroup based on the whole chloroplast genomes and the concatenated sequence of PCGs, respectively. Phylogeny showed that R. membranaceus was closely related to R. yunnanensis. These data enrich knowledge of Ranunculaceae genetics and will contribute to further studies of R. membranaceus in molecular breeding, genetic transformation, species identification, genetic engineering and phylogenetic research.

Two new ß-carboline alkaloids from the roots of Anemone altaica.

Two new ß-carboline alkaloids, anemonilins A and B (1-2), and two known ß-carboline alkaloids, flazine (3) and 4-(9H-ß-carbolin-l-yl)-4-oxo-butyric acid (4), were isolated from the roots of Anemone altaica. The structures of the isolated compounds were elucidated with spectroscopic and spectrometric methods (1D and 2DNMR, HRESIMS). Compounds 2 and 4 significantly attenuated the growth inhibition induced by lipopolysaccharide (LPS) in normal rat kidney tubule epithelioid (NRK52e) cells (p < 0.05 or p < 0.01). Furthermore, compound 2 significantly reduced the apoptosis (p < 0.05) and the caspase-3/9 expression of NRK52e cells induced by LPS.

Antinociceptive C19-diterpenoid alkaloids from the roots of Aconitum nagarum.

Three new C19-diterpenoid alkaloids, nagarumines A-C (1-3), together two known alkaloids, deoxyaconitine (4) and N-deethyldeoxyaconitine (5), were isolated from the roots of Aconitum nagarum. The structures of the new compounds were elucidated by spectral methods such as 1D and 2D (1H-1H COSY, HMQC, and HMBC) NMR spectroscopy, as well as high resolution mass spectrometry. The in vivo pharmacological studies revealed that nagarumine C (3) possessed comparable antinociceptive activity (ED50 = 76.0 µmol/kg) with the positive control drugs aspirin and acetaminophen.

Two new bis-diterpenoid alkaloids from Aconitum weixiense.

Further investigation on the roots of Aconitum weixiense led to the isolation of two new bis-diterpenoid alkaloids, named as weisaconitines E and F (1-2), which were elucidated by IR, HR-ESI-MS, 1D- and 2D-NMR analyses. Their structures are characterized as denudatine-atisine-type bis-diterpenoid alkaloids.

Dimeric benzylisoquinoline alkaloids from Thalictrum delavayi and their biological activities.

A phytochemical investigation of the whole plants of T. delavayi led to the isolation of five new dimeric benzylisoquinoline alkaloids, thalidelavines A-E (1-5), together with six known congeners (6-11). The structures and absolute configurations of new compounds were established based on analyses of spectroscopic data, ECD calculations, and single crystal X-ray crystallography. Thalidelavines A-E (1-5) were structurally complex bisbenzylisoquinoline alkaloids with various configurations. These isolated alkaloids were evaluated for their cytotoxic and immunosuppressive effects. Among them, both 9 and 10 displayed significant cytotoxicities against T98G cell lines with an IC50 value of 2.1 µM, compared with the positive CPT-11 (IC50 = 3.0 µM). In addition, 5-7 showed remarkable immunosuppressive effects. These findings not only enrich the structural diversity of bisbenzylisoquinoline alkaloids, but also provide potential candidates for the further development of the antitumor and immunosuppressive agents.

Grandiflolines A-F, new anti-inflammatory diterpenoid alkaloids isolated from Delphinium grandiflorum.

Delphinium grandiflorum L. (family Ranunculaceae), one of the most important and widely distributed Delphinium species, has received considerable interest due to its extremely high medicinal value. The discovery of novel metabolites from D. grandiflorum supported and broadened its application as an herbal medicine. In this study, the whole herb of D. grandiflorum was phytochemically investigated to obtain fourteen C19-lycaconitine-type diterpenoid alkaloids (1-14), including six undescribed alkaloids, grandiflolines A-F (1-6). The structural elucidation of them was accomplished by detailed spectroscopic analyses, mainly including HR-MS, 1D and 2D NMR (1H-1H COSY, NOESY, HMBC and HSQC), and IR spectra. New alkaloids 1-3 and 5 possess a characteristic △2,3 functional group in the A ring, while compounds 5 and 6 feature a rare OH-16 substituent. In addition, known compounds 7-12 were isolated from D. grandiflorum for the first time. Moreover, according to its medicinal use, new alkaloids 1-6 were estimated for their potential in vitro anti-inflammatory effects, and some of them exhibited inhibitory effects on NO production in LPS-activated RAW 264.7 macrophages. Our work enriched the chemical diversity of D. grandiflorum and the genus Delphinium and presented beneficial information for further investigations.

Characterization of the complete chloroplast genome of Clematis orientalis (Ranunculaceae).

The medicinal plant Clematis orientalis L. belongs to the family Ranunculaceae. In this study, we determined the complete chloroplast genome sequence of C. orientalis and its phylogenetic relationships with other species. The complete chloroplast genome of C. glauca is 159,518 bp in length, circular in structure, and has four regions including a large single-copy (LSC) region of 79,453 bp; a small single-copy (SSC) region of 17,997 bp; and two inverted repeat (IR) regions of 31,034 bp. The GC content of the genome is 38%, and those of LSC, SSC, and IR regions are 36.2, 31.4, and 42%, respectively. The genome encodes 129 unique genes, including 85 protein-coding genes, 36 tRNA genes, and eight rRNA genes. Phylogenomic analysis reveals that C. orientalis is most closely related to C. aethusifolia. This study contributes to better understanding of phylogenetic relationships of Ranunculaceae.

The ethnobotany, phytochemistry, and biological properties of Nigella damascena - A review.

This review is a systematic scientific work on medicinal and traditional use, on the chemical composition of specialized metabolites, volatile and non-volatile, on aspects related to toxicology and phytotherapy of Nigella damascena L. The genus Nigella (Ranunculaceae) is distributed throughout the Mediterranean basin, extending to northern India, and has been divided into three sections. Nigella damanscena L. is traditionally used as an ingredient in food, for example, as flavouring agents in bread and cheese, but is also known in folk medicine, used to regulate menstruation; for catarrhal affections and amenorrhea; as a diuretic and sternutatory; as an analgesic, anti-oedematous, and antipyretic; and for vermifuge and its disinfectant effects. This paper reviews the most dated to the latest scientific research on this species, highlighting the single isolated metabolites and exploring their biological activity. Fifty-seven natural compounds have been isolated and characterised from the seeds, roots, and aerial parts of the plant. Among these constituents, alkaloids, flavonoids, diterpenes, triterpenes, and aromatic compounds are the main constituents. The isolated compounds and the various extracts obtained with solvents of different polarities presented a diverse spectrum of biological activities such as antibacterial, antifungal, antitumour, antioxidant, anti-inflammatory, antipyretic, anti-oedema, and antiviral activities. Various in vitro and in vivo tests have demonstrated the pharmacological potential of ß-elemene and alkaloid damascenin. Unfortunately, the largest number of biological studies on this species and its metabolites have been conducted in vitro; therefore, further investigation is necessary to evaluate the toxicological aspects and real mechanisms of action of crude extracts to confirm the therapeutic potential of N. damascena.

The complete chloroplast genome of Clematis florida Thunb. (Ranunculaceae), an ornamental and medicinal plant from Henan province, China.

Clematis florida Thunb. is a herbaceous and perennial plant native to East Asia. The plant is resistant to cold but sensitive to heat. It is an ornamental and medicinal plant that has great commercial potential. Here, we assembled and characterized the complete chloroplast (cp) genome of C. florida. The cp genome of C. florida was characterized by Illumina pair-end sequencing and is 159,606 bp in total length. The genome includes a large single-copy (LSC) region of 79,467 bp, a small single-copy (SSC) region of 18,057 bp, and a pair of inverted repeats (IR) regions of 31,041 bp. The genome contains 135 genes including 91 protein-coding, 36 tRNA, and eight rRNA genes. Phylogenetic analysis based on 18 Clematis species indicates that C. florida is closely related to C. fusca in the Ranunculaceae. The phylogenetic relationships and taxonomic status of C. florida revealed by cp genome were consistent with the previous molecular studies, and can serve as a reference for future studies on molecular biology, evolution, and taxonomy in the genus Clematis.

New triterpenoid saponins from the whole plants of Clematis heracleifolia.

Three new triterpenoid saponins, heracleifolianosides A-C (1-3), together with seven known compounds (4-10), were isolated from the whole plants of Clematis heracleifolia. Moreover, three new secondary saponins (1a, 2a and 3a), two known secondary metabolites (5a and 7a) were obtained by alkaline hydrolysis. Their structures were elucidated by extensive spectroscopic analysis and chemical evidences. The cytotoxicity of eight native saponins and five prosapogenins against human breast tumor MDA-MB-231 and gastric carcinoma SGC-7901 cell lines were evaluated by MTT method. Remarkably, the prosapogenin monodesmosidic saponin 7a showed significant cytotoxicity against MDA-MB-231 or SGC-7901 cell lines with IC50 values in the range of 6.05-6.32 µmol/L. It is suggested that it might be a feasible way to change the inactive bisdesmosic triterpenoid saponins to active monodesmosic saponins by a simple procedure of alkaline hydrolysis.

Secondary Metabolites with Anti-Inflammatory from the Roots of Cimicifuga taiwanensis.

The genus Cimicifuga is one of the smallest genera in the family Ranunculaceae. Cimicifugae Rhizoma originated from rhizomes of Cimicifuga simplex, and C. dahurica, C. racemosa, C. foetida, and C. heracleifolia have been used as anti-inflammatory, analgesic and antipyretic remedies in Chinese traditional medicine. Inflammation is related to many diseases. Cimicifuga taiwanensis was often used in folk therapy in Taiwan for inflammation. Phytochemical investigation and chromatographic separation of extracts from the roots of Cimicifuga taiwanensis has led to the isolation of six new compounds: cimicitaiwanins A-F (1-6, respectively). The structures of the new compounds were unambiguously elucidated on the basis of extensive spectroscopic data analysis (1D- and 2D-NMR, MS, and UV) and comparison with the literature data. The effect of some isolates on the inhibition of NO production in lipopolysaccharide-activated RAW 264.7 murine macrophages was evaluated. Of the isolates, 3-6 exhibited potent anti-NO production activity, with IC50 values ranging from 6.54 to 24.58 µM, respectively, compared with that of quercetin, an iNOS inhibitor with an IC50 value of 34.58 µM. This is the first report on metabolite from the endemic Taiwanese plant-C. taiwanensis.

Anti-rheumatic, and analgesic effects by the parent tuberous roots of Aconitum jaluense in adjuvant induced arthritis rats.

AIM OF THE STUDY: The aim of this study was to test the anti-rheumatic effects of A. jaluense tubers in acute and chronic arthritis rats, and to assign its ingredients through UHPLC-TOF/MS. MATERIALS AND METHODS: Subcutaneous injection of carrageenan for acute arthritis and complete Freund's adjuvant (CFA) for chronic arthritis was carried out in the hind paw of SD rats. The paw volume was measured by a plethysmometer thermal hyperalgesia was tested using a thermal plantar tester, and mechanical hyperalgesia was evaluated by ankle flexion evoked vocalizations. The expression of c-Fos in the brain hippocampus was measured with the avidin-biotin-peroxidase technique. The ingredients were assigned by UHPLC-TOF/MS, chromatography was performed by UHPLC system with DAD detector and BEH C18 column, and spectroscopy was conducted by ESI-MS system. RESULTS AND DISCUSSION: The 80% ethanoic extract of A. jaluense tubers showed an acute anti-inflammatory effect by suppressing the edema volume in the hind paw of carrageenan-stimulated rats. In addition, A. jaluense tubers exerted an anti-rheumatic activity by reducing the secondary swelling volume from an immunological reaction in the left hind paw of CFA-induced chronic arthritis rats. Additionally, oral treatment with the 80% ethanoic extract -showed potent analgesic effects in the arthritis rats by recovering the paw withdrawal latency stimulated by the thermal hyperalgesia and by reducing the vocalization scores evoked by ankle flexion on both hind paws. Moreover, its treatment also indicated an anti-psychiatric effect by controlling the c-Fos protein expression of the brain hippocampus in CFA-stimulated arthritis rats. These results suggested that these therapeutic effects were exhibited by less toxic mono-esterified diterpenoid alkaloids (MDAs), and nontoxic non-esterified diterpenoid alkaloids (NDAs). CONCLUSION: A. jaluense tubers may act as viable therapeutic or preventive candidates for acute and chronic arthritis, particularly, for immune-inflammatory rheumatoid arthritis to suppress the pain and psychiatric condition.

The complete chloroplast genome sequence of Japanese buttercup Ranunculus japonicus Thunb.

Ranunculus japonicus is an important medicinal herb widely used in East Asia. In this study, we report the first complete chloroplast genome sequence of Ranunculus japonicus using next-generation sequencing technology. The chloroplast genome size of R. japonicus was 156,981 bp. A total of 129 genes were included, consisting 84 protein-coding genes, eight rRNA genes, and 37 tRNA genes. Thirteen protein-coding genes had intron (ycf3 gene, rps12 gene, rps12 gene, clpP gene contained two introns). A further phylogenomic analysis of Ranunculaceae, including 10 taxa, was conducted for assessing the placement of R. japonicus. It will provide valuable genetic information for this medicinally important species.