Combined transcriptomics and metabolomics to analyse the response of Cuminum cyminum L. under Pb stress.

Lead (Pb) can disrupt plant gene expression, modify metabolite contents, and influence the growth of plants. Cuminum cyminum L. is highly adaptable to adversity, but molecular mechanism by which it responds to Pb stress is unknown. For this study, transcriptomic and metabolomic sequencing was performed on root tissues of C. cyminum under Pb stress. Our results showed that high Pb stress increased the activity of peroxidase (POD), the contents of malondialdehyde (MDA) and proline by 80.03 %, 174.46 % and 71.24 %, respectively. Meanwhile, Pb stress decreased the activities of superoxide dismutase (SOD) and catalase (CAT) as well as contents of soluble sugars and GSH, which thus affected the growth of C. cyminum. In addition, Pb stress influenced the accumulation and transport of Pb in C. cyminum. Metabolomic results showed that Pb stress affected eight metabolic pathways involving 108 differentially expressed metabolites, primarily amino acids, organic acids, and carbohydrates. The differentially expressed genes identified through transcriptome analysis were mainly involved the oxidation reductase activity, transmembrane transport, phytohormone signaling, and MAPK signaling pathway. The results of this study will help to understand the molecular mechanisms of C. cyminum response to Pb stress, and provide a basis for screening seeds with strong resistance to heavy metals.

The role of cur ole of curcumin against paclitax cumin against paclitaxel-induced o el-induced oxidativ xidative stress and DNA damage in testes of adult male rats.

BACKGROUND: Paclitaxel is a widely used drug for the treatment of cancer, but it possesses toxic effects on male reproductive system. Administering paclitaxel with an antioxidant has become a strategy for preventing the side effects of paclitaxel. Although curcumin is an antioxidant, data concerning the effect of curcumin on paclitaxel-induced testis tissue are lacking. The present study was established to examine the protective impact of curcumin against testicular damage induced by paclitaxel. METHODS: In the study, 40 Wistar albino male rats were used and randomly divided into 4 groups (n:10). The control group received only saline solution; the curcumin group received curcumin throughout the experiment; the paclitaxel group received a total of four doses of paclitaxel on days 1, 7, 14, and 21 of the experiment; curcumin + paclitaxel group received curcumin throughout the experiment and a total of four doses of paclitaxel on days 1, 7, 14, and 21 of the experiment. At the end of the experiment, the rats were decapitated under xylazine and ketamine anesthesia and their testicles were removed. The sections obtained from the testicles were stained with Hematoxylin & Eosin and histopathological damage was evaluated. The TUNEL method was applied to determine apoptotic cells. Testosterone levels were measured in the blood serum. The Johnsen testicular biopsy score (JTBS) was used to evaluate testicular tubules. DNA damage was evaluated in sperm samples taken from the ductus epididymis using the comet assay technique. RESULTS: Testicular tissue was severely damaged in the paclitaxel group. In the curcumin + paclitaxel group, it was determined that the administration of curcumin with paclitaxel reduced the histological damage in the testicular tissue. Moreover, according to the JTBS, the value was significantly higher in the testicular tubules (p < 0.05). Testosterone levels were higher in curcumin + paclitaxel group than in paclitaxel group. DNA damage also decreased significantly in curcumin + paclitaxel group when compared to paclitaxel group (p < 0.05). DISCUSSION: The results showed that curcumin may be protective against damage caused by paclitaxel in the testicles of rats.

Effects of dietary supplementation of cumin (Cuminum cyminum L.) essential oil on expression of genes related to antioxidant, apoptosis, detoxification, and heat shock mechanism in heat-stressed broiler chickens.

This study was carried out to evaluate the impact of cumin essential oil (CEO) supplementation on levels of certain gene expression related to antioxidant, apoptotic, detoxific, and heat shock mechanisms in the breast meat and ileum of heat-stressed broilers. The study was conducted on a 2 × 6 factorial design (heat stress + feed additive) on 600 day-old male broiler chicks for a period of 42 days. From day 7 to 42, although broilers in heat stress groups (HT) were exposed to constant chronic heat stress (36 °C), others were housed at thermoneutral ambient temperature (TN). The chicks in both conditions were fed with 6 experimental diets: C0 (basal diet with no additive), ANTIB (basal diet + 100 mg/kg chloramphenicol), VITE (basal diet + 50 IU α-tocopherol), C2 (basal diet + 200 mg/kg CEO), C4 (basal diet + 400 mg/kg CEO), C6 (basal diet+ 600 mg/kg CEO). The results showed that heat stress upregulated (except for Bcl-2) the genes related to antioxidant, apoptosis, detoxification, and heat shock mechanism. However, cumin essential oil increased the dose-dependently positive effect on certain genes in tissues of the heat-stressed broilers and downregulated (except for Bcl-2) these genes.

Nigellidine (Nigella sativa, black-cumin seed) docking to SARS CoV-2 nsp3 and host inflammatory proteins may inhibit viral replication/transcription and FAS-TNF death signal via TNFR 1/2 blocking.

Tissue damage occurs in COVID-19 patients due to nsp3-induced Fas-FasL interaction/TNF-related apoptosis. Presently, possible therapeutic-drug, nigellidine against was screened by bioinformatics studies COVID-19. Atomic-Contact-Energy (ACE) and binding-blocking effects were explored of nigellidine (Nigella sativa L.) in the active/catalytic sites of viral-protein nsp3 and host inflammatory/apoptotic signaling-molecules Fas/TNF receptors TNFR1/TNFR2. A control binding/inhibition of Oseltamivir to influenza-virus neuraminidase was compared here. In AutoDock, Oseltamivir binding-energy (BE) and inhibition-constant (KI) was -4.12 kcal/mol and 959.02. The ACE values (PatchDock) were -167.02/-127.61/-124.91/-122.17/-54.81/-47.07. The nigellidine BE/KI with nsp3 was -7.61 and 2.66, respectively (ACE values were -221.40/-215.62/-113.28). Nigellidine blocked FAS dimer by binding with a BE value of -7.41 kcal/mol. Its strong affinities to TNFR1 (-6.81) and TNFR2 (-5.1) are demonstrated. Our present data suggest that nigellidine may significantly block the TNF-induced inflammatory/Fas-induced apoptotic death-signaling in comparison with a positive-control drug Oseltamivir. Further studies are necessary before proposing nigellidine as medical drug.

A nanotechnology-based new approach in the treatment of breast cancer: Biosynthesized silver nanoparticles using Cuminum cyminum L. seed extract.

The present study reports the anticancer activities of Cuminum cyminum L. (Cumin) seed extract, chemically synthetized silver nanoparticles (AgNPs) and biosynthesized silver nanoparticles (Bio-AgNPs) from Cumin seeds on human breast adenocarcinoma cell line (MCF-7) and human breast adenocarcinoma metastatic cell line (AU565). The synthetized nanoparticles were characterized by dynamic light scattering (DLS), UV-visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The cytotoxic and anticancer effects of AgNPs and Bio-AgNPs were determined by MTT assay. According to the cytotoxicity analysis, Bio-AgNPs appears to be less toxic against J774 macrophage cells than AgNPs since IC50 values were measured as 0.75 and 1.25 µg/ml for AgNPs and Bio-AgNPs, respectively. On the other hand, Bio-AgNPs demonstrated significant inhibitory effects on human breast cancer cells at non-toxic concentrations such as 0.25 and 0.5 µg/ml. However, at increased concentrations, the lethal effects of AgNPs on breast cancer cells were higher than Bio-AgNPs. When cytotoxic and anticancer characteristics of Cumin extract were investigated, it was established that it did not show any inhibitory effect on J774 cells, while killing the half of MCF-7 cells at investigated concentrations. Interestingly, Cumin extract gave rise to no inhibitory effects against AU565 cells. On the other hand, AgNPs and Bio-AgNPs exhibited considerable anticancer activities on both cell lines. The inhibition percentages of AgNPs on MCF-7 and AU565 cell lines were respectively evaluated as 95% and 97% at the highest concentrations applied (12.5 µg/ml). Similarly, we determined that 87.5% and 96% of MCF-7 and AU565 cells were respectively inhibited when they were exposed to the highest concentrations of Bio-AgNPs. Considering relatively toxic-free features of Bio-AgNPs prepared from Cuminum cyminum L. seed extracts, it can be thought that this formulation will be a pioneer in development of nanotechnology-based new anticancer drug for the treatment of breast cancer in near future.

Isolation, structural elucidation and immuno-stimulatory properties of polysaccharides from Cuminum cyminum.

The aim of the present study was to evaluate the effect of water-soluble polysaccharides from Cuminum cyminum to induce inflammatory response in immune cells and understand their underlying mechanisms. Weight average molecular weight (Mw) of polysaccharides varied between 191.4-512.2 × 103 g/mol. Polysaccharides induced RAW264.7 cells to release nitric oxide and express TNF-α, IL-1ß, IL-6 and IL-12 inflammatory cytokines. Polysaccharides activated NK-92 cells to produce TNF-α, IFN-γ, perforin, granzyme B, NKG2D and FasL. Activations of RAW264.7 and NK-92 cells were through NF-κB and MAPKs signal pathways indicated by the presence of phosphorylated NF-κB, ERK, JNK and p38 proteins. The polysaccharide structure was mainly constituted of →4)-Galp-(1→, →3)-Galp-(1→, →2)-Arap-(1→ and →2)-Arap-(1→ glycosidic linkages. Overall results suggested that polysaccharides from C. cyminum possessing lower MW and greater expanded conformation more effectively stimulate RAW264.7 and NK-92 cells and thus could be considered for further studies on their biomedical applications.

Yield, growth and Fe uptake of cumin (Cuminum cyminum L.) affected by Fe-nano, Fe-chelated and Fe-siderophore fertilization in the calcareous soils.

Cumin (Cuminum cyminum L.) from the Apiaceae family and as an important medicinal plant is greatly used for food production and medicinal purposes. The plant requires macro- and micro-nutrients including iron (Fe), which is not available under calcareous conditions. Accordingly, it was hypothesized the proper source (including the new methods of fertilization) and concentration of Fe can provide cumin with its required amounts of Fe under calcareous soils. The objectives were to determine: 1) the most efficient source of Fe fertilization (by spraying) on cumin yield and Fe uptake, and 2) the most optimum Fe concentration for cumin growth, yield production and seed fortification. A field experiment, as a completely randomized block design, with three replicates was conducted in the city of Aligudarz, Lorestan province, Iran. The cumin plants were sprayed twice during the season, before and after flowering (with a 10 day interval) according to the manufacturing Company. Three different types of Fe fertilization including Fe-nano-chelated (Fe-N), Fe-chelated (Fe-C) and Fe-siderophore (Fe-S) with the concentrations of 0 (control), 0.5 and 1 g/l were used for the experiment. Different plant parameters including grain Fe, crop yield, weight of 1000 grains, grain length, root length and stem length were determined. The highest yield of single plant was resulted by Fe-N1 (250 mg). Fe-N1 (3.8 g) and Fe-C1 (4.0 g) resulted in the highest weight of 1000 grains. However, the highest Fe concentration was related to treatment Fe-S1 (9.4 mg/kg). Plants treated with Fe-N (24.9 cm) and Fe-C (25.0 cm) treatments had the highest plant height. The highest root length was resulted by the Fe-N (8.9 cm) and Fe-C (9.1 cm) treatments. The control treatment resulted in the highest rate of root length/stem length (0.41). Treating the plants with Fe-C treatments resulted in the highest and significantly different grain length (6.8 mm). The concentration of 1 g/l of nano, chelated and siderophores were the most effective, significantly enhancing cumin yield and grain fortification. The results indicated the significant effects of Fe-N on plant yield and Fe uptake followed by the Fe-C method. The findings of this research work indicated that the Fe-N and the Fe-C methods were the most efficient methods enhancing cumin growth and yield. However, the most efficient method for seed fortification was the Fe-S method. The findings are of great nutritional, environmental and economical significance.

Protein Profiling of Non-model Plant Cuminum cyminum by Gel-Based Proteomic Approach.

INTRODUCTION: Cumin (Cuminum cyminum), a popular spice has been widely used in traditional medicine to cure various ailments. Despite the existence of scientific literature about its pharmacological properties, no successful proteome profiling has yet been attempted. OBJECTIVE: To optimise extraction of cumin proteins and analyse its profile by shotgun proteomics, using one-dimensional electrophoresis coupled with nano-ESI-LC-MS/MS. METHODOLOGY: As a first step, we have compared three extraction protocols for total proteins extraction from cumin. Extracted proteins were separated on one-dimensional gel and analysed by state-of-the-art linear ion trap (LTQ)-Orbitrap Velose and Q Exactive HF mass spectrometer. RESULTS: Evaluation of extraction method revealed significant differences in protein yield and proteome composition between the three extracts. LC-MS/MS allowed identification of several proteins with functional significance in various biological processes. CONCLUSION: This study provides identification of a large number of proteins and offers a molecular basis for future research on potential pharmacologically active cumin proteins. Copyright © 2017 John Wiley & Sons, Ltd.

Effects of growth regulators on callus induction and secondary metabolite production in Cuminum cyminum.

Cumin (Cuminum cyminum) is an annual plant from Apiaceae family that is cultivated in Iran as landraces. The most important chemical composition of the cumin essential oil was cuminaldehyde. In this research, the effect of different landraces and growth regulators was evaluated on callus induction, and best callus was used for amount of cuminaldehyde content. Node, root, leaf and hypocotyl explant from seedlings of Birjand and Qaen landraces were cultured on MS and MS5 medium supplemented with different concentrations of 2, 4-D and Kin. This experiment has been carried out in a completely randomised design with 3 replications. Percentage of callogenesis, callus volume, fresh and dry weight were measured. The best treatment for callus induction was 2.5 mg/L 2, 4-D and 0.5 mg/L Kin in MS5 medium. The best callus result was evaluated for cuminaldehyde content. An amount of 5.7% cuminaldehyde was measured using hydrodistillation method.

Development of a workflow for screening and identification of α-amylase inhibitory peptides from food source using an integrated Bioinformatics-phage display approach: Case study - Cumin seed.

The main objective of this study was to develop an efficient workflow to discover α-amylase inhibitory peptides from cumin seed. A total of 56 unknown peptides was initially found in the cumin seed protein hydrolysate. They were subjected to 2 different in silico screenings and 6 peptides were shortlisted. The peptides were then subjected to in vitro selection using phage display technique and 3 clones (CSP3, CSP4 and CSP6) showed high affinity in binding α-amylase. These clones were subjected to the inhibitory test and only CSP4 and CSP6 exhibited high inhibitory activity. Therefore, these peptides were chemically synthesized for validation purposes. CSP4 exhibited inhibition of bacterial and human salivary α-amylases with IC50 values of 0.11 and 0.04µmol, respectively, whereas CSP6 was about 0.10 and 0.15µmol, respectively. Results showed that the strength of each protocol has been successfully combined as deemed fit to enhance the α-amylase inhibitor peptide discovery.

Genetic linkage between protein and DNA polymorphisms and antioxidant capacity of Cuminum cyminum L. accessions.

This study aimed to link the genetic variation observed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and random amplified polymorphic DNA (RAPD) analysis among 11 Cuminum cyminum L. accessions, collected from diverse ecogeographical areas in Saudi Arabia, with their antioxidant capacity to better identify potential genotypes for breeding programs for this medicinal spice. SDS-PAGE analysis revealed genetic variation among cumin germplasms and distinct polymorphisms (100%). Protein polymorphisms were identified based on the number of polypeptide bands (288) with molecular weights ranging from 13.85 to 350 kDa, band intensity, the appearance of new bands, and the absence of other bands. RAPD analysis revealed 363 amplified DNA products with a high polymorphism value (98.88%) based on DNA band type (unique, non-unique, and monomorphic), DNA 90 to 1085-bp long, and band intensity. The unweighted pair group method with arithmetic mean clustering based on SDS-PAGE or RAPD and Jaccard's similarity coefficient divided cumin accessions into similar but distinct clusters with respect to their location of collection. The antioxidant potential of cumin accessions based on 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity, the ß-carotene-linoleate model system, and total phenolic and flavonoid contents revealed distinct variability. These data indicate that cumin is a valuable genetic resource with high antioxidant activity. Additionally, clustering based on antioxidant activity was not identical to that based on SDS-PAGE and RAPD. Data and clustering of SDS-PAGE and RAPD, combined with the high antioxidant capacity of cumin accessions, are important for the efficient use of genetic resources of cumin in breeding strategies and genetic improvement programs.

In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance.

Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas.

Physio-Biochemical Composition and Untargeted Metabolomics of Cumin (Cuminum cyminum L.) Make It Promising Functional Food and Help in Mitigating Salinity Stress.

Cumin is an annual, aromatic, herbaceous, medicinal, spice plant, most widely used as a food additive and flavoring agent in different cuisines. The study is intended to comprehensively analyse physiological parameters, biochemical composition and metabolites under salinity stress. Seed germination index, rate of seed emergence, rate of seed germination, mean germination time, plant biomass, total chlorophyll and carotenoid contents decreased concomitantly with salinity. In contrast, total antioxidant activity, H2O2, proline and MDA contents increased concurrently with stress treatments. Total phenolic and flavonoid contents were decreased initially about 1.4-fold at 50 mM, and thereafter increased about 1.2-fold at 100 mM NaCl stress. Relative water content remained unchanged up to 50 mM NaCl stress, and thereafter decreased significantly. About 2.8-fold electrolyte leakage was found in 50 mM, which increases further 4-fold at 100 mM NaCl stress. Saturated fatty acids (FAs) increased gradually with salinity, whereas unsaturation index and degree of unsaturation change arbitrarily along with the percent quantity of unsaturated FAs. Total lipid and fatty acid composition were significantly influenced by salinity stress. A total of 45 differentially expressed metabolites were identified, including luteolin, salvianolic acid, kaempferol and quercetin, which are phenolic, flavonoid or alkaloids in nature and contain antioxidant activities. Additionally, metabolites with bioactivity such as anticancerous (docetaxel) and antimicrobial (megalomicin) properties were also identified. The study evidenced that plant shoots are a rich source of metabolites, essential amino acids, phenolic compounds and fatty acids, which unveil the medicinal potential of this plant, and also provide useful insight about metabolic responses under salinity stress.

Water-deficit impact on fatty acid and essential oil composition and antioxidant activities of cumin (Cuminum cyminum L.) aerial parts.

This study is designed to examine the effect of water deficit on growth, fatty acid and essential oil composition, and antioxidant activities of Cuminum cyminum aerial part extracts. Plants were treated with different levels of water deficit: control (C), moderate water deficit (MWD), and severe water deficit (SWD). Plant growth (height, fresh and dry matter weights) as well as yield components were significantly increased under moderate water deficit and conversely reduced at severe level. Total fatty acid content decreased significantly with severity of constraint. Drought reduced considerably the proportions of major fatty acids and the unsaturated to saturated fatty acid ratio. The essential oil yield was 0.14% (based on the dry weight); it increased by 2.21-fold at MWD but decreased by 42.8% under SWD in comparison to the control. Drought results in the modification of the essential oil chemotype from 1-phenyl-1-butanol to 1-phenyl-1,2-ethanediol. Antioxidant activities of the acetone extracts were determined by two complementary test systems, namely, DPPH and ß-carotene/linoleic acid. The highest activity was exhibited by moderately stressed plants and was reduced significantly under SWD. In control plants, the total phenolic amount was 10.23 mg GAE/g DW, which increased by 1.5-fold under MWD and decreased by 42% under SWD.

RP-HPLC analysis of Jirakadyarishta and chemical changes during fermentation.

Jirakadyarishta, an Ayurvedic formulation prepared by the fermentation of a decoction of Cuminum cyminum (seeds) is traditionally used for intestinal disorders. RP-HPLC analysis of the decoction and the final processed formulation revealed that apigenin-7-O-[galacturonide (1 --> 4)-O-glucoside] and luteolin-4'-O-glucoside-7-O-galacturonide) were the two major constituents of the decoction of C. cyminum. Selective hydrolysis of 7-O-glucosides of luteolin and apigenin during fermentation resulted in an increase in the amount of luteolin and apigenin. The 4'-O-glucoside-7-O-galacturonide of luteolin and galacturonide derivative of apigenin were not hydrolyzed during fermentation. Monomeric phenolics, together with 5-hydroxymethyl furfural (5-HMF), were also introduced into the formulation through the jaggery and other plant materials during fermentation. This communication highlights the importance of the ancient processing methods used in Ayurveda.