Suspension culture of stem cells established of Calendula officinalis L.

Plant stem cell cultures have so far been established in only a few plant species using cambial meristematic cells. The presence of stem cells or stem cell-like cells in other organs and tissues of the plant body, as well as the possibility of de novo generation of meristematic cells from differentiated cells, allow to consider the establishment of stem cell cultures in a broader range of species. This study aimed to establish a stem cell culture of the medicinal plant Calendula officinalis L. Callus tissues were induced from leaf and root explants, and already at this stage, stem and dedifferentiated cells could be identified. The cell suspension cultures established both from the root- and leaf-derived calli contained a high proportion of stem cells (92-93% and 72-73%, respectively). The most effective combination of growth regulators for the development of stem cells in calli as well as cell cultures was 1.0 mg/L 2,4-D and 0.5 mg/L BAP. The highest amount of stem cells (5.60-5.72 × 105) was in cell suspension derived from the roots. An effective protocol for the establishment of marigold stem cell suspension culture was developed. The ratio of root-derived stem cells against dedifferentiated cells exceeded 90%.

Green and eco-friendly biosynthesis of zinc oxide nanoparticles using Calendula officinalis flower extract: Wound healing potential and antioxidant activity.

This study aimed to produce zinc oxide nanoparticles with Calendula officinalis flower extract (Co-ZnO NPs) using the green synthesis method. In addition, the antioxidant and wound healing potential of synthesized ZnO NPs were evaluated. The absorbance band at 355 nm, which is typical for ZnO NPs, was determined from the UV-Vis absorbance spectrum. The energy-dispersive X-ray spectroscopy (EDS) measurements revealed a high zinc content of 42.90%. The x-ray diffractometer data showed Co-ZnO NPs with an average crystallite size of 17.66 nm. The Co-ZnO NPs did not have apparent cytotoxicity up to 10 µg/mL (IC50 25.96 µg/mL). C. officinalis ZnO NPs showed partial cell migration and percent wound closure (69.1%) compared with control (64.8%). In addition, antioxidant activities of Co-ZnO NPs with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2 diphenyl-1 picrylhydrazil (DPPH) were evaluated and radical scavenging activity of 33.49% and 46.63%, respectively, was determined. These results suggest that C. officinalis extract is an effective reducing agent for the green synthesis of ZnO NPs with significant antioxidant and wound healing potential.

Study the Effect of Calendula officinalis Extract Loaded on Zinc Oxide Nanoparticle Cream in Burn Wound Healing.

The skin, the body's largest organ, acts as a protective barrier against pathogens and environmental damage. Skin burns can result from heat, chemicals, friction, or electricity. Nanoscience has recently been utilized to create ointments and creams for burns. Zinc oxide nanoparticles are crucial due to their antimicrobial and antioxidant properties. In this study, a cream containing nanoparticles was loaded with calendula extract, and its ability to promote tissue healing was investigated in Wistar rats with skin burns. The zinc oxide nanoparticles were chemically synthesized and loaded with calendula extract. The morphology and physicochemical properties of the nanoparticles were confirmed by SEM, ZETA size, XRD, and FTIR assays. The MTT technique was employed to assess the cream's impact on fibroblast growth. The antimicrobial activity of the nanoparticles was investigated against Pseudomonas using the MIC method. Real-time PCR was used to determine the expression of the Bax and Bcl-2 genes in aeruginosa. The results showed that zinc oxide nanoparticles at high concentrations increased the proliferation of the fibroblast cells. Histopathological studies showed granulation and epithelialization of the tissue without any hemorrhage or tissue infection during the first days of treatment with this cream. The animal models treated with the cream showed an increase in Bcl-2 gene expression and a decrease in Bax expression. We concluded that zinc oxide nanoparticles loaded with calendula extract have a practical effect in healing burn wounds due to their unique antibacterial properties of zinc oxide nanoparticles and their anti-inflammatory and wound-healing effects. The synergistic effect of these two substances significantly improved the healing process. This newly developed cream can be introduced as a successful and viable treatment option in burn wounds.

Exploring the Sequential-Selective Supercritical Fluid Extraction (S3FE) of Flavonoids and Esterified Triterpenoids from Calendula officinalis L. Flowers.

One of the many advantages of supercritical fluid extraction (SFE) is the possibility of using it in sequential and selective approaches. This is due to the use of a dynamic extraction mode in addition to the possibility of altering the composition of the modifier during the extraction process. In this study, the optimization of Calendula officinalis L. extraction of non-polar and polar compounds was achieved using three-level Box-Behnken designs (BBD). For non-polar compounds, the factors were pressure, temperature, and EtOH percentage. As for the polar compounds, the three variables were temperature, the total modifier percentage, and H2O added in the modifier as an additive. The recovery of selectively rich extracts in triterpendiol esters and narcissin was possible using a sequential two-step SFE. The first step was performed at 80 °C and 15% EtOH, and the second at 40 °C and 30% EtOH:H2O 80:20 v:v with a total of 60 min of extraction. Additionally, the SFE extraction of non-polar compounds was scaled up on a pilot-scale extractor, demonstrating similar results. Finally, the SFE results were compared to ultrasound-assisted extraction (UAE).

Effects of Calendula officinalis extract on liver histopathology, lipid profile, and oxidative stress in rats submitted to a diet rich in cholesterol and carbohydrates.

PURPOSE: To evaluate the modulatory properties of Calendula officinalis L. (Asteraceae) (C. officinalis) extract on cafeteria diet-fed rats. METHODS: A cafeteria diet was administered ad libitum for 45 days to induce dyslipidemia. Then, the rats were treated with the formulations containing C. officinalis in the doses of 50, 100, and 150 mg/kg or only with the vehicle formulation; the control group received a commercial ration. RESULTS: The cafeteria diet decreased glutathione S-transferase activity and high-density lipoprotein plasmatic levels and damaged the hepatic architecture. The C. officinalis extract was able to reduce lipid infiltration in liver tissue and to modulate oxidative stress and lipid profile markers. CONCLUSIONS: The correlations between the variables suggest a pathological connection between oxidative stress markers and serum lipid profile.

The potential of gamma irradiation on antioxidant capacity and genomic alterations in Calendula officinalis.

There are lines of evidence that ionizing radiations such as gamma rays can cause different biological effects on plants. Marigold (Calendula officinalis L.) is a member of the family Asteraceae. It possesses profound amounts of active ingredients. The aim of this study was to evaluate the changes imposed upon different dose levels of gamma radiation on some features of Calendula officinalis such as antioxidant activity, total phenolic compounds and flavonoid contents, antibacterial activity and genomic alterations. Calendula officinalis seeds were exposed to different doses of Gamma radiation (0, 10, 15, 20 and 25 GY). Total phenolics, flavonoids, antioxidant activity (measured by DPPH assay) using methanolic extracts of plants and antibacterial activity measured by the disc diffusion assay showed significant differences to the control samples. The samples treated with 10 GY gamma rays showed the highest total phenol and flavonoid contents. Antioxidant activity significantly differed between Gamma rays dose levels and it was the highest at 25 GY. Four bacterial strains including E. coli, Bacillus subtilis and Pseudomonas aeroginosa were used for the antibacterial assay. Extracts from plants treated with 25 GY gamma rays showed the highest antibacterial activity against the 4 bacterial strains. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to study the genetic variation. The polymorphism information content (PIC) for RAPD primers ranged from 3% to 13% and ranged from 6 to 13% for ISSR primers. Results indicated that ISSR markers were more efficient than RAPD markers, as they detected 25.57% polymorphic DNA bands compared to 21.31% polymorphism for RAPD markers.

Analysis of Lutein Content in Microencapsulated Marigold Flower Extract (Tagetes erecta L.) Using UHPLC-Q-Orbitrap-HRMS and Its Cytotoxicity in ARPE-19 Cells.

Organic solvents are commonly used to extract lutein. However, they are toxic and are not environmental-friendly. There are only a few reports on the quantification of lutein. Therefore, this study aimed to determine a suitable extraction method by which to obtain lutein from marigold flower (Tagetes erecta L.), using coconut oil to evaluate the cytotoxicity of extract in ARPE-19 cells, to optimize the encapsulation process for the development of microencapsulated marigold flower extract, and to develop the method for analysis of lutein by using UHPLC-Q-Orbitrap-HRMS. Coconut oil was used for the extraction of marigold flowers with two different extraction methods: ultrasonication and microwave-assisted extraction. The UHPLC-Q-Orbitrap-HRMS condition for the analysis of lutein was successfully developed and validated. Marigold flower extract obtained using the microwave method had the highest lutein content of 27.22 ± 1.17 mg/g. A cytotoxicity study revealed that 16 µM of lutein from marigold extract was non-toxic to ARPE-19 cells. For the development of microencapsulated marigold extract, the ratio of oil to wall at 1:5 had the highest encapsulation efficiency and the highest lutein content. Extraction of lutein using coconut oil and the microwave method was the suitable method. The microencapsulated marigold extract can be applied for the development of functional ingredients.

Modifications in steroid and triterpenoid metabolism in Calendula officinalis plants and hairy root culture in response to chitosan treatment.

BACKGROUND: Chitosan, a deacetylated derivative of chitin, is one of the most preferred biopolymers for use as biostimulants and biofertilizers in organic agriculture and as elicitors to enhance the productivity of plant in vitro cultures. Valued as a non-toxic, biodegradable, and environment-friendly agent, it is widely applied to improve plant growth and yield, the content of bioactive specialized metabolites, and resistance to stress conditions and pathogens. However, the influence of chitosan on the growth-defense trade-off, particularly the interplay between steroid and triterpenoid metabolism, has not been extensively investigated. RESULTS: In this study, Calendula officinalis pot plants and hairy root cultures exposed to chitosan treatment displayed reduced biomass and altered steroid and triterpenoid metabolism. Biosynthesis and accumulation of free forms of sterols (particularly stigmasterol) were inhibited, while the content of sterol esters increased remarkably. The content of some triterpenoids (mainly free triterpenoid acids) was slightly enhanced; however, the biosynthesis of triterpenoid saponins was negatively affected. CONCLUSIONS: These results indicate that in certain plants, chitosan treatment might not positively influence the growth and metabolite production. Therefore, to avoid unexpected effects, initial studies of the conditions of chitosan treatment are recommended, including the dose and the number of chitosan applications, the type of treatment (e.g., foliar or soil), and the vegetative stage of the treated plants.

Prenylated acetophenones from the roots of Calendula officinalis and their anti-inflammatory activity.

Calendula officinalis is a medicinal plant in the Asteraceae family, and it has a broad range of biological activities. In this study, we focused on the roots of C. officinalis, which have remarkable anti-inflammatory properties. By using a bioassay-guided fractionation approach, prenylated acetophenones 1 and 2-of which 1 was previously unknown-were isolated, and their structures were determined by spectroscopic analysis. Both compounds decreased lipopolysaccharide-stimulated NO production in J774.1 cells. This study could lead to the use of the Calendula roots as a natural source of inflammatory mediators.

In Vitro α-Amylase and α-Glucosidase Inhibitory Effects, Antioxidant Activities, and Lutein Content of Nine Different Cultivars of Marigold Flowers (Tagetes spp.).

Marigolds (Tagetes spp.) are major sources of bioactive compounds. The flowers are used to treat a variety of illnesses and have both antioxidant and antidiabetic effects. However, marigolds exhibit a wide range of genetic variations. Because of this, both the bioactive compounds and biological activities of the plants differ between cultivars. In the present study, nine marigold cultivars grown in Thailand were evaluated for their bioactive compound content, as well as for their antioxidant and antidiabetic activities, using spectrophotometric methods. The results showed that the Sara Orange cultivar possessed the highest total carotenoid content (431.63 mg/100 g). However, Nata 001 (NT1) had the highest amount of total phenolic compounds (161.17 mg GAE/g), flavonoids (20.05 mg QE/g), and lutein (7.83 mg/g), respectively. NT1 exhibited strong activities against the DPPH radical and ABTS radical cation, and had the highest FRAP value as well. Moreover, NT1 demonstrated the most significant (p < 0.05) α-amylase and α-glucosidase inhibitory effects (IC50 values of 2.57 and 3.12 mg/mL, respectively). The nine marigold cultivars had reasonable correlations between lutein content and the capacity to inhibit α-amylase and α-glucosidase activities. Hence, NT1 may be a good source of lutein; it may also be beneficial in both functional food production and medical applications.

Hydrological performance of rain gardens having Calendula officinalis plant with varied planting mixtures.

The rain gardens (RGs) have been one of the best management practices in cities to reduce the impact of urban flooding. However, very little is known about various design parameters of RGs, viz., the type of plantation, planting mixtures, and RG dimensions. This study pertains to examining the influence of planting mixtures on the variations of percolation rates of the RG with Calendula officinalis plant and without plants. Six types of planting mixtures in different experimental RGs have been tried. It has been observed that the percolation rate increases with a higher percentage of compost in the planting mixture for RGs with and without plants. The percolation rate is highest for the planting mixture having 25% compost. The runoff rate reduces with a higher percentage of compost in the planting mixture for RGs with C. officinalis and bare surfaces. No runoff is produced in RGs with plant having a compost of more than 20% in the planting mixture. The outcome of the study will be useful in deciding the composition of the planting mixture which will keep the RG plant healthy and at the same time improve the hydrological performance leading to lowering urban flooding magnitude.

Effects of Calendula officinalis and Capsicum annum glycolic extracts on planktonic cells and biofilms of multidrug-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa.

Empirical knowledge of natural plant extracts is increasingly proving to be a promising field. The effect of Calendula officinalis L. (CO) and Capsicum annum (CA) glycolic extracts (GlExt) have potential that should be further developed in microbial tests. The effect of CO-GlExt and CA-GlExt was evaluated on eight multidrug-resistant clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa, as well as collection strains for each bacterial. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined in comparison with 0.12% chlorhexidine. The tests were performed on single species biofilms, at 5 min and 24 h, using the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. The MIC and MBC of the extract ranged from 1.56 to 50 mg mL-1 for all strains evaluated. Analysis of the MTT assay revealed a strong antimicrobial potential of CA-GlExt, comparable to chlorhexidine. The findings suggest that CA-GlExt is effective against multidrug-resistant strains of K. pneumoniae and P. aeruginosa in planktonic state and biofilms.

Burkholderia cepacia CS8 improves phytoremediation potential of Calendula officinalis for tannery solid waste polluted soil.

Microbes have shown potential for the bioremediation of tannery waste polluted soil. During our previous study, it was observed that heavy metal resistant Burkholderia cepacia CS8 augmented growth and phytoremediation capability of an ornamental plant. Objective of the present research work was to evaluate the capability of B. cepacia CS8 assisted Calendula officinalis plants for the phytoremediation of tannery solid waste (TSW) polluted soil. The TSW treatment significantly reduced growth attributes and photosynthetic pigments in C. officinalis. However, supplementation of B. cepacia CS8 which exhibited substantial tolerance to the TSW amended soil, augmented growth traits, carotenoid, proline, and antioxidant enzymes level in C. officinalis under toxic and nontoxic regimes. Inoculation of B. cepacia CS8 augmented plant growth (shoot length 13%, root length 11%), physiological attributes (chlorophyll a 14%, chlorophyll b 17%), antioxidant enzyme activities (peroxidase 24%, superoxide dismutase 31% and catalase 19%), improved proline 36%, phenol 32%, flavonoids 14% and declined malondialdehyde (MDA) content 15% and hydrogen peroxide (H2O2) level 12% in C. officinalis at TSW10 stress compared with relevant un-inoculated plants of TSW10 treatment. Moreover, B. cepacia CS8 application enhanced labile metals in soil and subsequent metal uptake, such as Cr 19%, Cd 22%, Ni 35%, Fe 18%, Cu 21%, Pb 34%, and Zn 30%, respectively in C. officinalis plants subjected to TSW10 stress than that of analogous un-inoculated treatment. Higher plant stress tolerance and improved phytoremediation potential through microbial inoculation will assist in the retrieval of agricultural land in addition to the renewal of native vegetation.

During the current study, it was observed that combination of Calendula officinalis and metal tolerant Burkholderia cepacia CS8 not only improved plant growth but also helped phyto-extraction of pollutants present in the tannery solid waste polluted soil. According to our information, research work describing the phytoremediation potential of native metal tolerant microbes and ornamental plants has not been reported in Pakistan.

Heterologous Production of Calendic Acid Naturally Found in Calendula officinalis by Recombinant Fission Yeast.

Calendic acid (CA) is a conjugated fatty acid with anti-cancer properties that is widely present in seed oil of Calendula officinalis. Using the co-expression of C. officinalis fatty acid conjugases (CoFADX-1 or CoFADX-2) and Punica granatum fatty acid desaturase (PgFAD2), we metabolically engineered the synthesis of CA in the yeast Schizosaccharomyces pombe without the need for linoleic acid (LA) supplementation. The highest CA titer and achieved accumulation were 4.4 mg/L and 3.7 mg/g of DCW in PgFAD2 + CoFADX-2 recombinant strain cultivated at 16 °C for 72 h, respectively. Further analyses revealed the accumulation of CA in free fatty acids (FFA) and downregulation of the lcf1 gene encoding long-chain fatty acyl-CoA synthetase. The developed recombinant yeast system represents an important tool for the future identification of the essential components of the channeling machinery to produce CA as a high-value conjugated fatty acid at an industrial level.

Phytoextraction of nickel, lead, and chromium from contaminated soil using sunflower, marigold, and spinach: comparison of efficiency and fractionation study.

Heavy metals in soil pose a serious threat through their toxic effect on the human food chain. Phytoremediation is a clean and green potentially cost-effective technology in remediating the heavy metal-contaminated soil. However, the efficiency of phytoextraction is very often limited by low phytoavailability of heavy metals in soil, slow growth, and small biomass production of hyper-accumulator plants. To solve these issues, accumulator plant(s) with high biomass production and amendment(s) which can solubilize metals in soil is required for better phytoextraction. A pot experiment was conducted to assess the efficiency of phytoextraction of sunflower, marigold, and spinach as affected by the incorporation of Sesbania (solubilizer) and addition of gypsum (solubilizer) in nickel (Ni)-, lead (Pb)-, and chromium (Cr)-contaminated soil. A fractionation study was conducted to study the bioavailability of the heavy metals in contaminated soil after growing the accumulator plants and as affected by using soil amendments (Sesbania and gypsum). Results showed that marigold was the most efficient among the three accumulator plants in phytoextraction of the heavy metals in the contaminated soil. Both sunflower and marigold were able to reduce the bioavailability of the heavy metals in the post-harvest soil, which was reflected in their (heavy metals) lower concentration in subsequently grown paddy crop (straw). The fractionation study revealed that carbonate and organically bound fractions of the heavy metals control the bioavailability of the heavy metals in the experimental soil. Both Sesbania and gypsum were not effective in solubilizing the heavy metals in the experimental soil. Therefore, the possibility of using Sesbania and gypsum for solubilizing heavy metals in contaminated soil is ruled out.

Catechol-O-methyltransferase Inhibitors from Calendula officinalis Leaf.

Calendula officinalis is commonly known as marigold and its flowers are used in herbal medicines, cosmetics, perfumes, dyes, pharmaceutical preparations, and food products. However, the utility of its leaves has not been studied in depth. The purpose of the present study was to identify the major compounds in C. officinalis leaves and to determine the inhibitory properties of the isolated compounds toward human catechol-O-methyltransferase (COMT), a key neurotransmitter involved in Parkinson's disease and depression. We isolated and identified ten compounds, including two phenylpropanoids and seven flavonoids, from C. officinalis leaf extracts, of which four flavonoids were identified from C. officinalis leaves for the first time. Eight compounds exhibited COMT inhibitory activities with IC50 values of less than 100 µM. Our results indicate that compounds in C. officinalis leaves are potentially effective for preventing Parkinson's disease and depression. Thus, C. officinalis leaves may hold promise as dietary supplements.

The volatile organic compounds and palatability of mixed ensilage of marigold (Tagetes erecta L.) crop residues.

With increasing acreage of cash crops, the use of their by-products as supplements for livestock feed becomes an important factor. Marigold (Tagetes erecta L.) account for more than half of the world's loose flower production. However, there is no precedent for the abundantly available marigold crop residue (MCR) being used as feed in agricultural production, probably because of its strong pungent taste. This study aimed to evaluate the biotransformation of the volatile organic compounds (VOCs) of MCR by mixed ensilage and assess its palatability by cattle. Caryophyllene, the most prevalent VOC in MCR, decreased by 29.11% (P < 0.05), 38.85% (P < 0.05), 37.15% (P < 0.05), and 28.36% (P < 0.05) ensilage with corn meal (CM), bran (BR), crop corn (CC), and straw (ST), respectively. The acetic acid content increased by 686.05% (P < 0.05), 1337.21% (P < 0.05), 1244.19% (P < 0.05), and 1795.34% (P < 0.05) after mixed ensilage with CM, BR, CC, and ST, respectively. The total amount of alcoholic VOCs followed an overall increasing trend during mixed storage and 10 new alcohols were obtained. Over seven days, feed intake of mixed ensilage MCR by cattle differed significantly (P < 0.05) among treatments compared with MCR and was highest in MCRCM. Combined with palatability trials, the best MCR feed intake was achieved with MCRCM. The findings shed light on how feed odor can be improved and how degradation of terpenes can be enhanced in practical applications by mixed ensilage.

Wound healing and antibacterial capability of electrospun polyurethane nanofibers incorporating Calendula officinalis and Propolis extracts.

In order to produce a healing accelerator antibacterial wound dressing, different electrospun polyurethane (PU)-based nanofibers inclusive Calendula officinalis and Propolis ethanolic extracts were fabricated. The measurement of minimum inhibitory concentrations (MIC) against Methicillin-resistant Staphylococcus aureus (MRSA) determined the concentrations of incorporating extracts. Then the morphological properties of the produced polyurethane (PU), polyurethane/C. officinalis (PU/CO), polyurethane/Propolis (PU/PR), polyurethane/C. officinalis/Propolis (PU/CO/PR) were analyzed by scanning electron microscopy (SEM). The physicochemical features and biological characteristics of the fabricated nanofibers were evaluated. Subsequently, the antibacterial and wound-healing efficiency of electrospun wound dressings were tested under in vivo situation. The electrospun PU/CO/PR nanofiber illustrated the most degree of antibacterial, antioxidant, and cell proliferation efficiencies. In vivo examination and histological analysis confirmed significant improvement in the complete, well-organized wound-healing process in MRSA-infected wounds treated with PU/CO/PR. These outcomes described PU/CO/PR electrospun nanofibers as a wound dressing that can significantly facilitate wound healing with notable antibacterial, antioxidant, and cell proliferation properties.

Accumulation and distribution characteristics of rare earth elements (REEs) in the naturally grown marigold (Tagetes erecta L.) from the soil.

Rare earth elements (REEs) are considered environmental pollutants that have received extensive attention recently. The accumulation of REEs in plants is important because REEs can eventually enter the human body via the food chain. Marigolds are widely utilized as medicinal and commercial plants in medicine, feed, and therapeutics. Due to the extremely high demand for marigold in global, it is urgent to investigate the accumulation and distribution of REEs in marigold plants to reduce human and animal health risks. Marigold leaves tended to bioaccumulate the highest amounts of REEs from soil compared with other tissues. The distribution patterns of REEs in marigold were similar to those in the rhizosphere soil, which was enriched in light rare earth elements. Cerium accumulated most in marigold and soil, accounting for nearly 50% of ΣREEs, followed by lanthanum, neodymium, and yttrium. Roots were the most susceptible tissue affected by soil REE concentration, and a significant positive correlation was observed for REEs in the roots of marigold and soils (R = 0.87), while no significant correlation was observed for REEs in soils and other tissues. REEs were poorly transferred from soil to marigold, with bioaccumulation factor values for all tissues of marigold less than one. Additionally, REEs exhibited a positive correlation with Al and Fe in the roots, stems, leaves, and flowers of marigold. The present research revealed the biological interactions between marigold and soil and the distribution of REEs in various parts of marigold. It provides a reference for large-scale commercial cultivation and potential environmental risk in the future.

Growth, biochemical, and antioxidant response of pot marigold (Calendula officinalis L.) grown in fly ash amended soil.

The present study was carried out to determine the impact of FA application on growth performance, biochemical parameters, and antioxidant defense activity of Calendula officinalis. The results revealed that under a low dose of FA (40%) amended soil, the plant growth performance and metal tolerance index (MTI) were increased compared to control plants and further decreased with increased FA application (60%, 80%, and 100% FA). In addition, the incorporation of 40% FA in soil not only improved the physicochemical properties of soil but also increased the biochemical parameters in the Calendula plant, however, these parameters declined under high FA applications. It was also observed that antioxidant enzyme activity (SOD, CAT, POD, and APX) in leaves of Calendula officinalis increased at high FA application (100% FA) to combat heavy metal stress from FA. The overall study suggests that 40% FA amended soil is the best suitable dose for growing Calendula officinalis and can be considered as metal tolerant species for phytoremediation of 40% FA amended soil.Novelty statement: Fly ash (FA) management is a major problem nowadays. The present study was carried out for FA utilization and to determine the impact of FA amended soil on growth performance, antioxidant properties, and biochemical attributes of Calendula officinalis. This is a sustainable approach in which waste (FA) utilization was done simultaneously with the enhancement in response of the medicinally potent Calendula species. The novelty of this study also suggests that Calendula has phytoremediation potential for remediation of heavy metal polluted soil. Further, the relationship between the growth, biochemical parameters, and antioxidant defense mechanism of Calendula grown on FA amended soil was studied which has not been studied so far. It was found that Calendula is a hyperaccumulator that can adapt to heavy metal stress from FA due to its ability to mitigate oxidative damage. Statistical analysis (ANOVA, Duncan's multiple range test, and PCA) was done for the results obtained using SPSS (11.5) and Origin 8 Pro software.