Phytochemical Characterization Research Articles

Bioactive phytochemicals from leaf extract of Erlangea tomentosa S. Moore (Bothriocline longipes (Oliv. & Hiern.) used to treat bacterial infections in Uganda

BackgroundThe leaves of Erlangea tomentosa (ET) are used to treat several bacterial infections like respiratory tract infections, skin infections, diarrhea and gastrointestinal disorders. However, there is limited information about the compounds responsible for its bacterial activities. The study aimed at isolation and characterisation of phytochemicals from the leaf extracts of ET.ResultsThree compounds were isolated from leaf extracts of ET and characterised. Two compounds; eriodictyol-7-O-β-glucoside (1) and 5, 7, 3′, 4′-tetrahydroxy-7-O-[6"-O-(acetyl)-β-D-glucopyrancsyloxy]-flavanone (coccinoside A) (2) were identified from ethyl acetate extract while one compound; di-(2-ethylhexyl) phthalate (3) was identified from dichloromethane extract.ConclusionCompounds; 1, 2 and 3 were isolated and identified from ET and genus Erlangea for the first time. The reported antibacterial properties of compound 3 in literature could be responsible for the antibacterial activities of ET and the traditional use of ET.

Phytochemical Characterization and Evaluation of Antioxidant Activity in Differently Polished Proso Millet

Phytochemical Characterization and Evaluation of Antioxidant Activity in Differently Polished Proso Millet

Phytochemical Analysis, Isolation, and Characterization of Gentiopicroside from Gentiana kurroo and Cytotoxicity of Biosynthesized Silver Nanoparticles Against HeLa Cells.

Gentiana kurroo Royle, a critically endangered Himalayan herb, is valued in treating leucoderma, syphilis, bronchial asthma, hepatitis, etc. The current investigation performed quantitative and qualitative phytochemical analysis of G. kurroo root extracts prepared in chloroform, methanol, and ethyl acetate. The phenolic and flavonoid contents were the highest in methanol and chloroform extract, respectively. Several pharmacologically important compounds were identified through gas chromatography-mass spectrometry. Antioxidant analysis revealed methanolic extract to be the most efficient scavenger of 2,2-diphenyl-1-picrylhydrazyl (IC50 = 114µgmL-1), hydrogen peroxide (IC50 = 109.9µgmL-1), and superoxide (IC50 = 74.63µgmL-1) radicals. Gentiopicroside was isolated from the methanolic root extract through silica-gel column-chromatography, and the characterization of concentrated fractions was achieved employing various analytical techniques. Pertaining to silver nanoparticle (GkAgNPs) synthesis, different physicochemical parameters were optimized and it was observed that root extract treated with silver-nitrate (0.5mM) at 60°C and incubated in dark for at least 120min after initial color change, yielded GkAgNPs optimally. GkAgNPs were anisotropic and polydisperse and exhibited characteristic surface plasmon resonance (424nm), crystalline face-centered cubic geometry, size (50-300nm), and zeta-potential (- 16.3mV). FT-IR spectra indicated the involvement of phenols and flavonoids in AgNPs synthesis. GkAgNPs were evidenced as strongly cytotoxic (IC50 = 1.964µgmL-1) against HeLa cells and also showed deformed cellular morphology, a significant reduction in viable cell counts and colony-forming efficiency (4.08%). The findings suggest potential applications in drug development for treating serious human diseases. To the best of our knowledge, this study represents the first report on the isolation of gentiopicroside, the bio-fabrication of GkAgNPs using G.kurroo root extract, and their strong bioefficacy against HeLa cells.

Antimicrobial Activity of Biohardened Plant Extracts of Albizia Amara

Albizia amara is an important medicinal plant commonly found in dry forests of India. It belongs to leguminaceae and its common name in telugu is Nallaregoo (or) Chigaraku. All of the plant parts i.e., roots, shoots, leaves, flowers, seeds and bark, were exclusively used in traditional medicine for curing different diseases like diarrhea, skin diseases gonorrhea, poisonous bites and leprosy. Due to its wide pharmacological activities and presence of several therapeutic constituents, Albizia amara has been selected for tissue culture studies in the present investigation. A systemic approach for increasing the survival rate of tissue culture raised A.amara plantlets using bioinoculants was explored . The studies reported that biohardening of tissue cultured plantlets with mixed inoculum of Pseudomonas fluorescens and Trichoderma viride enhanced plant survival rate (82 %). This microbial association helped in inducing systemic resistance in plants to overcome abiotic and biotic stress caused during acclimatization. The phytochemical studies of biohardened plants exhibited high antimicrobial activity due to the presence of more secondary metabolites. These secondary metabolites probably due to their antagonistic properties gave protection against pathogenic attack during transplantation. Phytochemical analysis of different plant extracts like, ethyl acetate, chloroform, methanolic, and petroleum ether extracts were studied. The ethyl acetate extract of dual inoculant treated plants showed maximum antimicrobial activity. In bacteria, it showed maximum zone of inhibition on B.cereus (11.5±1.73), followed by E.coli (10±0.82 mm) . In fungi, it showed maximum inhibition on F.oxysporum (13.25±0.28mm) followed by C.albicans (8.5±0.17mm). The above strategy helped in overall improvement of morphological, physiological and phytochemical characteristics of tissue culture plantlets.

Phytochemical characterization and evaluation of anti-amnesic activity of two source plants of Shankhapushpi

BackgroundShankhapushpi is an important Ayurvedic drug used for treating various disease conditions of nervous system. Convolvulus prostratus Forssk. is the genuine source plant for Shankhapushpi as per Ayurvedic Pharmacopoeia of India; however Clitorea ternatea L. is widely used as Shankhapushpi in southern part of India. In this study, comparative phytochemical and pharmacological evaluation has been done in two plants such as Convolvulus prostratus and Clitorea ternatea used as Shankhapushpi. MethodsPhytochemical comparison was done by High Performance Thin Layer Chromatography (HPTLC) and High Performance Liquid Chromatography (HPLC). Detailed metabolite profiling was performed using Q-TOF-LC/MS-MS analysis. Antiamnesic activity of selected plants has been evaluated against scopolamine induced amnesia model in Wistar rats. ResultsPhytochemical studies showed that only a few chemical constituents are common for both species. Most of the phytochemicals are different for selected species. LC/MS analysis showed presence of genipin and 7-hydroxyflavone in both species. Pharmacological study showed that both plants possess antiamnesic activity against Scopolamine induced memory loss; However C. ternatea possesses significant antiamnesic activity than that of C. prostratus. ConclusionThe study provided valuable information about the selected species in terms of its phytochemical composition and pharmacological properties. The study also provided a scientific support for using both species as Shankhapushpi.

Extraction and characterization of phytochemicals from Bauhinia variegata flowers using ultrasound and microwave techniques

This research investigates the efficacy of three distinct extraction techniques; ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), and conventional extraction (CE) for the isolation of phytochemicals from the flowers of Bauhinia variegata. The findings demonstrated that both UAE and MAE surpass CE, yielding higher concentrations of bioactive compounds while significantly reducing the extraction time. UAE demonstrated the highest extraction efficiency, followed by MAE. The UAE yielded significantly higher levels of total phenolic content (43.97 mg GAE/g), total flavonoid content (24.08 mg QE/g), total anthocyanin content (16.37 mg CGE/g), DPPH antioxidant activity (19.29 mg TE/g), and FRAP antioxidant activity (16.75 mg TE/g) compared to MAE and conventional extraction methods. GC-MS analysis revealed a diverse array of phytochemicals, including flavonoids, phenolic compounds, and terpenoids, with some compounds showing higher concentrations in UAE. Reverse phase HPLC profiling further confirmed the enhanced extraction of phenolic compounds using UAE, including gallic acid, caffeic acid, coumaric acid, quercetin, and kaempferol. This study reveals the efficiency, time-saving, and eco-friendliness of UAE for extracting valuable bioactive compounds from B. variegata flowers. The findings optimize extraction processes in the food, pharmaceutical, and nutraceutical industries.

Phytochemical Screening, Isolation and Characterization of Bioactive Compounds from Ethanol Extract of Memecylon lushingtonii Gamble

The primary goal of this study was to isolate and characterize the bioactive compounds present in the M. lushingtonii Gamble leaves using ethanol as solvent for the extraction using Soxhlet apparatus for 8 h. Based on the chemical analysis, column chromatography using different solvents depending on polarity further isolated the components. Isolation of the extract was carried out using toluene and ethyl acetate solvents with varying concentrations and showed two single spots by TLC at the rentation time (Rt) of 0.66 and 0.72 for the solvent’s toluene and ethyl acetate (8:12 and 2:18), respectively. Various spectroscopy techniques (UV, NMR, IR, and MASS) were employed for the characterisation of the compounds and elucidation of their structures. The results confirmed the presence of alkaloids, phenolics, flavonoids, tannins, sterols and terpenoids. The spectroscopic analysis revealed the compounds were quercetin (Rf: 0.66) and luteolin (Rf: 0.72) which were both flavonoid group and contains five and four –OH groups, respectively in their basic structure (C6-C3-C6). The results demonstrated that the leaves of M. lushingtonii Gamble exhibited a higher concentration of polyphenolic chemicals, which contribute to the diverse therapeutic properties.

Chemical Characterization and Antioxidant Activity of Rosa canina L. From Çekerek Region of Yozgat, Turkey

Rosehip, renowned for its bioactive richness, including phenolic acids, flavonoids, carotenoids, and ascorbic acid, exhibits significant antioxidant and health-promoting potential. This study investigated the physicochemical, mineral, and phytochemical characteristics of rosehips sourced from the Çekerek region of Yozgat, Turkey, to evaluate their suitability as a functional food. The rosehips exhibited high dry matter content and low water activity, suggesting excellent stability for storage and processing. Furthermore, the rosehips contained substantial mineral content, including potassium, calcium, magnesium, iron, and zinc, which enhances their nutritional value. Phytochemical analysis revealed a significant phenolic content, potent antioxidant activity, and exceptional vitamin C levels, positioning Çekerek rosehips as a superior natural antioxidant source. These findings underscore the influence of Çekerek's unique environmental conditions on rosehip composition and highlight their potential applications in the functional food, nutraceutical, and pharmaceutical sectors, addressing the increasing demand for natural antioxidants. This research contributes to the understanding of regional variations in Rosa canina and provides a basis for further investigation into the bioavailability, clinical applications, and development of value-added products from Çekerek rosehips.

Improved Skin Barrier Function Along with Hydration Benefits of Viola yedoensis Extract, Aesculin, and Schaftoside and LC-HRMS/MS Dereplication of Its Bio-Active Components

The skin hydration level is a key factor that influences the physical and mechanical properties of the skin. The stratum corneum (SC), the outermost layer of the epidermis, is responsible for the skin’s barrier function. In this study, we investigated the role of a unique composition of Viola yedoensis extract for its ability to activate CD44, a cell-surface receptor of hyaluronic acid, and aquaporin-3, a water-transporting protein, in human keratinocytes (HaCaT). An ELISA assay evaluating the protein expression levels of CD44, aquaporin-3 (AQP3), filaggrin, and keratin-10 revealed that V. yedoensis extract upregulated the levels of CD44 and AQP3 by 15% and 78%, respectively. Additionally, V. yedoensis extract demonstrated a comparative effect on water vapor flux in TEWL and lipid perturbation in DSC versus the reference, glycerin. In light of this new biological efficacy, a detailed phytochemical characterization was undertaken using an integrated LC-HRMS/MS-based metabolomics approach, which provided further insights on the chemistry of V. yedoensis. This led to the identification of 29 secondary metabolites, 14 of which are reported here for the first time, including esculetin, aesculin, apigenin and kaempferol C-glycosides, megastigmane glycosides, roseoside, platanionoside B, and an eriojaposide B isomer, along with the rare, calenduloside F and esculetin diglucoside, which are reported for the first time from the genus, Viola. Notably, two active components identified in the V. yedoensis extract, namely, aesculin and schaftoside, showed an upregulation of the protein expression of CD44 in HaCaT cells by 123% and 193% within 24 h of treatment, respectively, while aesculin increased AQP3 levels by 46%. Aesculin and schaftoside also significantly upregulated the expression of K-10 levels by 299% and 116%, which was considerably higher than sodium hyaluronate, the positive control. The rationale used to characterize the new structures is outlined along with the related biosynthetic pathways envisioned to generate roseoside and Eriojaposide B. These findings provide new molecular insights to deepen the understanding of how V. yedoensis extract, along with the biomarkers aesculin and schaftoside, restores the skin barrier and skin hydration benefits.

Phytochemical Characterization and Antibacterial Activity of Carthamus Caeruleus L. Aqueous Extracts: In Vitro and In Silico Molecular Docking Studies.

In order to valorize natural resources and the traditional use of medicinal plants in Algeria, this study exploits the antibacterial effect of Carthamus caeruleus L. Since there are few studies on this plant despite its notable therapeutic potential, this work aims to characterize the chemical composition of Carthamus caeruleus L. leaf and root aqueous extracts and to evaluate their antibacterial activity through an in vitro and in silico studies. Spectrophotometric assays and HPLC results revealed 22 components in the roots and 16 in the leaves. Disc diffusion and microdilution methods were used to study the antibacterial properties against nine standard bacterial strains. The results showed that roots exhibited the best activity on most tested strains. Both extracts were also able to inhibit the growth of Staphylococcus aureus ATCC 25923 and Escherichia coli ATSC 25922. Furthermore, no nucleic acid leakage or membrane damage was detected. However, molecular docking of the molecules indicates that some constituents have significant affinity and stability for DNA gyrase. Gallic acid, luteolin, myricetin, and orientin were found to have the highest score. The molecular docking data suggest, for the first time, that the antibacterial activity may be caused by the inhibition of DNA gyrase.

Phytochemical and pharmacological overview on Paerida foetida L. and Sphaeranthus amarnthoides Burm.f.: A Review

The phytochemistry and pharmacological activities of the plants Paerida foetida L. and Sphaeranthus amaranthoides Burm.f. are the goals of this review. The current study's findings are supported by the review literature and published research on the pharmacological and phytochemical characteristics of Sphaeranthus amaranthoides Burm.f. and Paerida foetida L. These plants are useful for antibacterial, anthelmintic, antioxidant, cytotoxic, anti-diabetic, hypolipidaemic, and hepatoprotective effects. The plant extracts were regarded as safe for animals according to toxicity studies. Both plants contain a variety of phytoconstituents, including tannins, terpenoids, steroids, phenols, flavonoids, and saponins, each of which has a distinct pharmacological action. For many years, people have been exploring the natural world to find novel phytoconstituents that have been employed in the treatment of various illnesses. It also offers a way to discover novel semisynthetic and synthetic molecules as well as new natural entities. This review article examines the claims, traditional knowledge, and phytochemical and pharmacological analyses of these plants, as well as their potential uses in the future.

Preparation and Phytochemical Characterization of Grewia Asiatica Fruit Extract by Successive Extraction Method

The present study aimed on the preparation and phytochemical characterization of Grewia asiatica fruit extract using the successive extraction method. Grewia asiatica, commonly known as Phalsa, is a tropical fruit rich in bioactive compounds with potential health benefits. The extraction process involved successive use of solvents with increasing polarity petroleum ether, benzene, chloroform, ethyl acetate and methanol. Each extract was subjected to qualitative phytochemical screening to identify and quantify various secondary metabolites, including alkaloids, flavonoids, tannins, phenols, saponins, and glycosides. The results indicated that the different extracts exhibited varying phytochemical profiles, with methanol extract showing the highest concentration of flavonoids, tannins, and phenols.

Phytochemical characterization and evaluation of anti-inflammatory and anticancer activities of Cistus laurifolius originated propolis

Phytochemical characterization and evaluation of anti-inflammatory and anticancer activities of Cistus laurifolius originated propolis

Phytochemical Characterization and Assessment of Antioxidant and Anti-Alzheimer Effects of Algerian Seseli Tortuosum.

HPLC analysis, phytochemical screening, thin layer chromatography, polyphenols and flavonoid contents were conducted to determine the bioactive contents of the Algerian Seseli tortuosum plant. Antioxidant activity was tested using DPPH and ABTS scavenging assays, reducing power, phenanthroline and silver nanoparticle (SNP) assays. The anti-Alzheimer's activity of extracts was also performed in vitro and in silico. Phytochemical analysis highlighted the richness ofthe extracts in terms of coumarins, terpenoids and quercetin. The quantitative determination of total polyphenols and flavonoids showed that the highest amounts occurredinthe dichloromethane (DCME) and methanolic (MeOH) extracts. The antioxidant activities indicated a moderate potential. Compared with galantamine, DCME had a significantly greater anti-Alzheimer effect on BChE(CI50=9.14±1.74 μg/ml and 34.75±1.99 μg/ml respectively). An in silicostudy of anti-Alzheimer activity revealed a significant effect of quercetin(-30,13 KJ/mol). Conclusion: This study demonstrated the richness of the phytochemical components of seseli tortuosum,which are responsible for several biological properties, mainly their anti-Alzheimer potential.

Phytochemical and Biochemichal Profile of Raw Extracts and Oil from Curcuma longa Rhyzomes Grown in Diabo (Côte d'Ivoire)

Aims: The aim of this study was to determine the phytochemical and biochemichal profile of Curcuma longa grown in Diabo, central Côte d'Ivoire. Place and Duration of Study: Department of Biochemistry-Genetics, Unit for Training and Research in Biological Sciences, Peleforo University GON COULIBALY Korhogo, BP 1328 Korhogo; The first part of this study was carried out from April 1, 2023 to December 31, 2023; the second part was conducted from March 01, 2024 to July 31, 2024. Methodology: Several phytochemical tests based on colorimetric reactions were used to perform qualitative and quantitative analyses of polyphenols, flavonoids and tannins. To characterize the oil and crude extracts, the indices of iodine and refractive as well as pH and Brix were determined. Results: The results show the presence of several secondary metabolites in the crude extracts, including polyphenols (53.1 ± 2.08 mg Eq AG/g extract), flavonoids (2.7 ± 0.22 mg Eq AG/g) and catechic tannins (9.1 ± 0.7 mg Eq C/g) in variable quantities. Biochemical analyses showed a pH close to neutral (pH = 6.68). The Brix value (23.20 ± 0.26 min), shows that Curcuma longa extracts, in particular EHCL, contain several soluble solids such as reducing sugars, acids, salts, pectins and nitrogenous matter. As far as the oil is concerned, the iodine value obtained was 84.1 ± 0.1 g I2 /100 g fat; this high value is close to 100 g. The refractive index was 1.52 ± 0.61. Conclusion: Phytochemical and biochemical characteristics show that Curcuma longa rhizomes could have interesting pharmacological potential.

Phytochemical characterization, toxicity and pharmacological profile of the central effects of the fixed fruit pulp oil of Mauritia flexuosa L.F. (buriti)

Anxiety and depression are mental disorders that have been exponentially increasing over the last decades. Psychopharmacology emerged to try to alleviate the symptoms of these disorders; however, the side effects and the time it takes to achieve the desired effect are factors that decrease the search for and adherence to treatment. To remedy this situation, new compounds capable of improving the performance of these medications and reducing their adverse effects have been sought. The use of medicinal plants has been widely employed for this purpose. Mauritia flexuosa F.L., a palm tree with high incidence in Brazil, has been heavily targeted as all its parts are usable. The objective of this study is to evaluate the effects of fixed oil from the fruit of the buriti palm in models of depression and anxiety. The phytochemical profile of this oil and its toxicity were also investigated. The experiments conducted included the open field, rotarod, forced swim, and elevated plus maze tests. As a result, it was observed that the fixed oil from buriti palm presented 18 compounds, with elaidic acid being the major one, and showed no signs of toxicity. However, it demonstrated a possible stimulating activity in the open field test and had no effect on the motor system in the rotarod test. Furthermore, it exhibited an antidepressant-like effect in the forced swim test but an anxiety-like effect in the elevated plus maze test. Therefore, buriti oil may potentially be used in new formulations to assist in the treatment of anxiety and depression.

Corema album Berry Juice as a Protective Agent Against Neurodegeneration.

Background/Objectives: Corema album berries are edible fruits from the Iberian Atlantic coast, characterized by a rich polyphenolic composition, which endows their juice with potential protective effects against neurodegeneration. This study aimed to evaluate the potential of the relatively lesser-known C. album berries as a novel neuroprotective agent against neurodegenerative diseases. Methods: The phenolic compounds of the juice were characterized using UHPLC-HRMS (Orbitrap). The SH-SY5Y neuroblastoma line was used to determine the preventive effect of the juice against H2O2-induced oxidative stress. Furthermore, neuronal cells were differentiated into dopaminergic and cholinergic lines and exposed to 6-hydroxydopamine and okadaic acid, respectively, to simulate in vitro models of Parkinson's disease and Alzheimer's disease. The ability of the juice to enhance neuronal viability under toxic conditions was examined. Additionally, its inhibitory effects on neuroprotective-related enzymes, including MAO-A and MAO-B, were assessed in vitro. Results: Phytochemical characterization reveals that 5-O-caffeoylquinic acid constitutes 80% of the total phenolic compounds. Higher concentrations of the juice effectively protected both differentiated and undifferentiated SH-SY5Y cells from H2O2-induced oxidative damage, reducing oxidative stress by approximately 20% and suggesting a dose-dependent mechanism. Moreover, the presence of the juice significantly enhanced the viability of dopaminergic and cholinergic cells exposed to neurotoxic agents. In vitro, the juice inhibited the activity of MAO-A (IC50 = 87.21 µg/mL) and MAO-B (IC50 = 56.50 µg/mL). Conclusions: While these findings highlight C. album berries as a promising neuroprotective agent, further research is required to elucidate its neuroprotective mechanisms in cell and animal models and, ultimately, in human trials.

Isolation and characterization of phytochemicals from leaf extract of Memecylon lushingtonii Gamble and their evaluation for biological properties

Memecylon lushingtonii Gamble (MLG) belongs to the Melastomataceae family and is used in Ayurvedic and Siddha medicine. Several Memecylon species have been recorded to be utilized by tribal in the treatment of various maladies. This plant is not explored scientifically except for the preliminary phytochemicals, antioxidant, and antimicrobial activities in the methanol leaves extract (Sivu et al., 2013). Discovery of new phytochemicals may help in drug discovery. The present study was an attempt to determine the phytochemical constituents in MLG leaves ethyl acetate (EtOAc) extract and to isolate bioactive compounds and evaluate their potential therapeutic applications. Ethyl acetate extract was evaluated for total phenolics and flavonoids; phytochemical constituents, antibacterial and antioxidant activities. Further, bioactive compound from 4th fraction of column chromatography was isolated and characterized by HPLC,LC-MS and H1NMR. The results indicated 37.5 µg/mL of phenolics, 60 µg/mL of flavanoids and twenty-one bioactive compounds by GCMS– analysis. The extract demonstrated considerable antibacterial activity with different concentrations like 50, 100 and 150 µg against Staphylococcus aureus, Bacillus licheniformis, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. Highest antibacterial activity was found against K.pneumoniae (Zone of inhibition was 11 mm) at 150 µg .In addition, the EtOAc extract displayed free radical scavenging activity (IC50= 39.83 µg/ml). The isolated compound from 4th fraction of column chromatography was characterized as a triterpenoid, ursolic acid. In conclusion, our findings show that M. lushingtonii Gamble leaf EtOAc extract contains triterpenoid, ursolic acid, and also, many types of active compounds with therapeutic activities, particularly antibacterial and free radical scavenging activity.

Isolation, Identification, Phytochemical Characterization, and In Vitro Anti-Inflammatory Property of an Endophytic Trametes Versicolor CL-1 Isolated from Rosa Roxburghii.

This study explores the anti-inflammatory potential of an endophytic fungus, Trametes versicolor CL-1, isolated from the fruit tissues of Rosa roxburghii. Morphological and molecular analyses confirmed the identity of CL-1. An ethyl acetate extract (CL-E) from its fermentation broth was subjected to UPLC-HRMS and GNPS molecular networking. The analysis revealed a diverse array of secondary metabolites, including 11 terpenes, 7 flavonoids, 10 cinnamic acid derivatives, 6 oligopeptides, and 9 fatty acids, as verified by LC-MS/MS. Notably, CL-E exhibited significant in vitro anti-inflammatory activity in RAW264.7 cells. Furthermore, molecular docking studies predicted favorable binding interactions of key compounds 1 within CL-E with the NLRP3 inflammasome (PDB ID: 6NPY). These findings suggest T. versicolor CL-1 as a promising source of natural anti-inflammatory agents and unveil R. roxburghii as a potential reservoir for discovering novel bioactive metabolites.

Genetic and phytochemical evaluation of M2 generation mutants of fenugreek (Trigonella foenum-graecum L.) induced by gamma rays and Ethyl Methane Sulphonate (EMS).

Fenugreek (Trigonella foenum-graecum L.) is highly esteemed for its therapeutic properties and is widely used in traditional medicine and modern pharmacology. Enhancing its genetic traits and phytochemical profile, particularly its trigonelline content, can significantly increase its medicinal and agricultural value. This study aims to investigate the effects of gamma rays and Ethyl Methane Sulphonate (EMS) as mutagenic agents on the genetic and phytochemical characteristics of the M2 generation of fenugreek, focusing on genetic diversity and desirable trait enhancement. To achieve this, various concentrations of EMS and gamma rays were administered to fenugreek seeds, and 27 traits were assessed in the resulting M2 generation. These traits were analyzed for variance, mean values, and correlations. The genetic diversity of 23 M2 offspring was investigated using nine Start Codon Targeted (SCoT) markers. The genetic diversity assessment involved Principal Coordinate Analysis (PCoA) and cluster analysis, utilizing the Dice similarity coefficients and the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). A Bayesian model provided deeper insights into the genetic structure. Results revealed that lower doses of gamma rays (100Gy) and EMS (0.2%) positively impacted specific traits. In comparison, higher doses (200Gy and 0.4% EMS) increased seed trigonelline content to 0.71mg/g dry weight. Among the SCoT markers, SCoT-9 was the most efficient, segregating the populations into three clusters. The first three principal components in the PCoA explained 20% of the total variance, leading to seven subgroup populations distinction. These findings underscore the potential of induced mutagenesis in enhancing desirable traits in fenugreek.