Influence of calcium channel modulators on the production of serotonin, gentisic acid, and a few other biosynthetically related phenolic metabolites in seedling leaves of salt tolerant rice variety Nonabokra.

Rice, a most salt-sensitive cereal plant, adopts diverse pathways to withstand sodium chloride-induced salinity-related adversities. During the present study, attempt was made to understand the role of calcium on metabolite profile of the leaves of salt tolerant rice seedlings of variety of Nonabokra under sodium chloride induced salinity, by Gas Chromatography-Mass Spectrometry-based metabolomics approach. Calcium availability in the seedlings was reduced or enhanced applying inhibitors (vanadyl sulfate, lanthanum chloride, and verapamil) or promoters of calcium influx (calcimycin also known as calcium ionophore A23187) in the sodium chloride (100 mM) supplemented growth medium. Growth medium of ten-day-old seedlings was replaced by sodium chloride supplemented hydroponic solution with promotor or inhibitors of calcium channel. Fifteen days old seedlings were harvested. It was observed that depletion of calcium availability increased the level of serotonin and gentisic acid whereas increased calcium level decreased these metabolites. It was concluded from the results that production of the signaling molecules serotonin and gentisic acids was elevated in calcium-deficient seedlings under salt stress the condition that was considered as control during the experiment. The two signaling molecules probably help this tolerant rice variety Nonabokra to withstand the salt-induced adversities.

Influence of sodium chloride on growth and metabolic reprogramming in nonprimed and haloprimed seedlings of blackgram (Vigna mungo L.).

Salinity hinders agricultural productivity worldwide by distressing plant metabolism. Growth of blackgram (Vigna mungo L. var. Sulata), an adverse climate-resistant pulse, is arrested under salinity. Present research integrates study of physio-biochemical parameters and non-targeted metabolomics approach to explore the alterations in metabolic pathway during adaptive responses of nonprimed and haloprimed blackgram seedlings grown hydroponically under NaCl stress. Salinity provoked accumulation of peroxides, compatible solutes and phenolics which increased free radical scavenging activities of nonprimed seedlings under salinity. Pre-germination seed halopriming abrogated NaCl-mediated adversities in haloprimed plantlets favouring better growth. Thus, farmers may adopt seed halopriming technique to improve blackgram productivity in saline-prone fields. Additionally, metabolomics study uncovered numerous metabolites amongst which 35 compounds altered significantly under salinity. The candidate metabolites were aspartic acid, L-glutamic acid, L-proline, L-asparagine, DL-isoleucine, L-homoserine, citrulline, L-ornithine, D-altrose, D-allose, N-acetyl-D-mannosamine, fructose, tagatose, sucrose, D-glucose, maltose, glycerol-1-phosphate, D-sorbitol, benzoic acid, shikimic acid, 4-hydroxycinnamic acid, arbutin, succinic acid, pipecolic acid, fumaric acid, nicotinic acid, L-pyroglutamic acid, oxalic acid, glyceric acid, maleamic acid, adenine, guanosine, lauric acid, stearic acid and porphine. Comparing metabolic responses of nonprimed and haloprimed seedlings, it was clear that efficient alteration in carbohydrate metabolism, phenolics accumulation, amino acid, organic acid and nucleic acid metabolism were the key places of metabolic reprogramming for tolerating salinity. Overall, we report, for the first time, 35 contributory candidate compounds that constituted core fundamental metabolome invoking salinity tolerance in nonprimed and haloprimed blackgram. These metabolites may be targeted by biotechnologists to produce high vigour salt-tolerant transgenic blackgram via genetic engineering.

Metabolite profiling and in-vitro colon cancer protective activity of Cycas revoluta cone extract.

The methanolic extract of Cycas revoluta cone (MECR) was analyzed by GC-MS and UHPLC for metabolite profiling and was evaluated for anti-colon cancer property by using in vitro assays like Cell Viability Assay, Colony Formation Assay, ROS Determination, Flowcytometry, DAPI staining assay, Tunel assay. GC-MS and HPLC analysis confirmed the presence of different phytochemicals in the extract of Cycas revoluta cone. In-vitro studies showed MECR extract showed significant anti-colon cancer activity by reducing proliferation and inducing apoptosis in colon cancer cell (HCT-8) line, but no such activity was seen in normal colon cell (CCD-18Co) line. The investigation confirms that MECR may be a promising candidate in colon cancer protection.

Phytochemical composition, ß-glucuronidase inhibition, and antioxidant properties of two fractions of Piper betle leaf aqueous extract.

RATIONALE: Piper betle leaf, used as masticatory in South Asia, is also medicinally important. OBJECTIVE: This work was done to analyze phytochemical composition of two solvent fractions (chloroform and ethyl acetate) of the aqueous extracts obtained from eight varieties of P. betle leaves and to identify the active components against ß-glucuronidase by chemometric analysis. RESULTS: Twenty-four phenolic compounds, in addition to different organic acids, fatty acids, amino acids, sugars, and polyols, were identified from the solvent fractions. The extracts inhibited the enzyme ß-glucuronidase. Piceatannol was the most active constituent against the enzyme (activity 12 times higher than that of silymarin), Chlorogenic acid also inhibited ß-glucuronidase (activity 4.4 times higher when compared to silymarin). 2,2'-Diphenyl-1-picrylhydrazyl and superoxide free radical scavenging activities of both the fractions of eight varieties of P. betle leaf extracts showed very strong antioxidant potentiality. CONCLUSION: The findings validated some medicinal properties of the said leaves. PRACTICAL APPLICATIONS: Edible leaves of Piper betle are medicinally and economically important. Leaves of different local varieties are reported to be used for the treatment of different diseases. The leaves have many biological properties, hepatoprotection being one of them. A large number of rural population is economically dependent on the cultivation of betel vine. But with a rapid change in lifestyle, the chewing habit of P. betle is decreasing ultimately affecting the livelihood of farmers dependent on betel cultivation. Knowledge on ß-glucuronidase inhibitory activity and the mechanism for hepatoprotection of different P. betle varieties may validate the medicinal properties of betel, which would increase consumption of these leaves.

Metabolomics analysis of Cajanus cajan L. seedlings unravelled amelioration of stress induced responses to salinity after halopriming of seeds.

Soil salinity has become a major concern for agriculture. Such constraints not only reinforce the urgent need to understand the underlying mechanisms by which plants cope during salt stress but also to develop cost-effective and farmer friendly halopriming technique to alleviate the adverse effects of salinity to some extent. Metabolomics approach was used to explore different responses to physiological metabolites and pathway variations that occur during salt stress responses in Cajanus cajan L. var. Rabi and to understand the role of halopriming in ameliorating stress at the level of metabolite. Seedlings raised from non-primed and haloprimed seeds, grown in hydroponic solution, were subjected to different concentrations of NaCl. After 21 days, metabolites were extracted, derivatized and analyzed by GC-MS. The data were analysed by different multivariate analyses. Chemometric study of the identified metabolites indicated that the leaves responded most to NaCl induced stress than the stem and root with production of beta-cyano-L-alanine and also increased level of different compatible solutes. O-Acetylsalicylic was also found to increase in all the parts upon facing stress but, such upregulated metabolite production was downregulated in the leaves when the seeds were haloprimed before germination, although many of the metabolites, including beta-cyanoalanine, showed a trend of increase with increase in salt concentrations. Important metabolites produced by C. cajan seedlings in response to salinity were unravelled. Pre-germination haloprimimg of seeds resulted in amelioration of NaCl induced stress, as the levels of stress induced metabolites were lowered.

GC-MS based metabolite profiling and angiotensin I-converting enzyme inhibitory property of black tea extracts

Abstract Angiotensin I-converting enzyme inhibitors are used as therapeutic agents for the treatment of hypertension. Regular consumption of black tea (Camellia sinensis (L.) Kuntze, Theaceae) has been reported to lower blood pressure. The aims of the present work were to compare chemical composition and angiotensin I-converting enzyme inhibitory properties of infusion and decoction of four samples of black tea. GC/MS based metabolomics approach helped in identification of fifty-one metabolites including ten organic acids, one inorganic acid, sixteen amino acids, two sugars, five sugar alcohols, fifteen phenols and flavonoids, two fatty acids from infusions and decoctions of four black tea samples. Partial least squares discriminant analysis and orthogonal partial least squares discriminant analysis models showed good classification among the two groups, diffusion and infusion, based on metabolites. Both infusion and decoction inhibited the enzyme. However, the activity differed with samples. Multivariate analysis also segregated extracts on the basis of activity. Thearubigin, theaflavin, catechin inhibited the enzyme. Epicatechin, epigallocatechin gallate, gallic acid, caffeine showed lower activity.

Differential responses of cell wall bound phenolic compounds in sensitive and tolerant varieties of rice in response to salinity.

In plants, cell wall bound phenolics change in response to stress. The aim of the study was to investigate the effect of NaCl induced stress on wall bound phenolics in four rice varieties, of which two (Bhutnath, Nonabokra) were salt tolerant and two (MTU 7029, Sujala) were salt sensitive. After germination, seedlings were grown in hydroponic solution and subjected to salinity stress (25 mM, 50 mM, 100 mM and 150 mM NaCl) on day 12. Wall bound phenolic compounds were determined by GC-MS based metabolite analysis. Total seven wall bound phenols were identified from the leaf tissues and eight from the root tissues. Ferulic acid and 4-hydroxycinnamic acid were found in all the four varieties. After NaCl treatment, these two wall bound phenols increased in the leaves of tolerant varieties only. Significant inverse correlation between leaf length and leaf fresh weight with wall bound ferulic acid and 4-hydroxycinnamic acid in Nonabokra suggests the positive role of these wall bound phenolics in salt tolerance.

Metabolomics analysis of rice responses to salinity stress revealed elevation of serotonin, and gentisic acid levels in leaves of tolerant varieties.

A GC-MS based analytical approach was undertaken to understand the metabolomic responses of seedlings of 2 salt sensitive (Sujala and MTU 7029) and 2 tolerant varieties (Bhutnath, and Nonabokra) of indica rice (Oryza sativa L.) to NaCl induced stress. The 4 varieties responded differently to NaCl treatment with respect to the conserved primary metabolites (sugars, polyols, amino acids, organic acids and certain purine derivatives) of the leaf of rice seedlings. However, there were significant differences in salt induced production of chorismic acid derivatives. Serotonin level was increased in both the salt tolerant varieties in response to NaCl induced stress. In both the salt tolerant varieties, increased production of the signaling molecule gentisic acid in response to NaCl treatment was noticed. Salt tolerant varieties also produced increased level of ferulic acid and vanillic acid. In the salt sensitive varieties, cinnamic acid derivatives, 4-hydroxycinnamic acid (in Sujala) and 4-hydroxybenzoic acid (in MTU 7029), were elevated in the leaves. So increased production of the 2 signaling molecules serotonin and gentisic acid may be considered as 2 important biomarker compounds produced in tolerant varieties contributing toward NaCl tolerance.

Metabolic profile and β-glucuronidase inhibitory property of three species of Swertia

ABSTRACT β-Glucuronidase inhibitors are suggested as potential hepatoprotective agents. Swertia chirayita (Roxb.) Buch.-Ham. ex C.B. Clarke, Gentianaceae, is known for its hepatoprotective and anti-hepatotoxic activity in Ayurvedic system of medicine for ages. This plant is substituted by other species like S. decussata Nimmo ex C.B. Clarke and S. bimaculata (Siebold & Zucc.) Hook. f. & Thomson ex C.B. Clarke. The aim of the study was to compare metabolite profile and β-glucuronidase inhibitory activity of these three important species of Swertia and to identify the active constituents. S. chirayita (IC50 210.97 µg/ml) and S. decussata (IC50 269.7 µg/ml) showed β-glucuronidase inhibitory activity significantly higher than that of silymarin, the known inhibitor of the enzyme. The activity of S. bimaculata was low. The metabolites present in the three species were analyzed by HPLC and GC-MS based metabolomics approach. Five amino acids, twenty one organic acids, one inorganic acid, eight fatty acids, twenty one phenols including xanthones, eight sugars, seven sugar alcohols, five terpenoids and amarogentin were identified. Activities of the xanthones mangiferin (IC50 16.06 µg/ml), swerchirin (IC50 162.84 µg/ml), decussatin (IC50 195.11 µg/ml), 1-hydroxy-3,5,8-trimethoxy xanthone (IC50 245.97 µg/ml), bellidifolin (IC50 390.26 µg/ml) were significantly higher than that of silymarin (IC50 794.62 µg/ml). Quinic acid (IC50 2.91 mg/ml), O-acetylsalicylic acid (IC50 48.4 mg/ml), citric acid (IC50 1.77 mg/ml), D-malic acid (IC50 14.82 mg/ml) and succinic acid (IC50 38.86 mg/ml) also inhibited the enzyme β-glucuronidase. The findings suggest that constituents, in addition to the xanthones, probably also contribute to the bioactivity of different Swertia species by synergistic effect. Further in vivo study is required to support the claim.

Sansevieria roxburghiana Schult. & Schult. F. (Family: Asparagaceae) Attenuates Type 2 Diabetes and Its Associated Cardiomyopathy.

BACKGROUND: Sansevieria roxburghiana Schult. & Schult. F. (Family: Asparagaceae) rhizome has been claimed to possess antidiabetic activity in the ethno-medicinal literature in India. Therefore, present experiments were carried out to explore the protective role of edible (aqueous) extract of S. roxburghiana rhizome (SR) against experimentally induced type 2 diabetes mellitus (T2DM) and its associated cardiomyopathy in Wistar rats. METHODS: SR was chemically characterized by GC-MS analysis. Antidiabetic activity of SR (50 and 100 mg/kg, orally) was measured in high fat diets (ad libitum) + low-single dose of streptozotocin (35 mg/kg, intraperitoneal) induced type 2 diabetic (T2D) rat. Fasting blood glucose level was measured at specific intermissions. Serum biochemical and inflammatory markers were estimated after sacrificing the animals. Besides, myocardial redox status, expressions of signal proteins (NF-κB and PKCs), histological and ultrastructural studies of heart were performed in the controls and SR treated T2D rats. RESULTS: Phytochemical screening of the crude extract revealed the presence of phenolic compounds, sugar alcohols, sterols, amino acids, saturated fatty acids within SR. T2D rats exhibited significantly (p < 0.01) higher fasting blood glucose level with respect to control. Alteration in serum lipid profile (p < 0.01) and increased levels of lactate dehydrogenase (p < 0.01) and creatine kinase (p < 0.01) in the sera revealed the occurrence of hyperlipidemia and cell destruction in T2D rats. T2DM caused significant (p < 0.05-0.01) alteration in the biochemical markers in the sera. T2DM altered the redox status (p < 0.05-0.01), decreased (p < 0.01) the intracellular NAD and ATP concentrations in the myocardial tissues of experimental rats. While investigating the molecular mechanism, activation PKC isoforms was observed in the selected tissues. T2D rats also exhibited an up-regulation in nuclear NF-κB (p65) in the cardiac tissues. So, oral administration of SR (50 and 500 mg/kg) could reduce hyperglycemia, hyperlipidemia, membrane disintegration, oxidative stress, vascular inflammation and prevented the activation of oxidative stress induced signaling cascades leading to cell death. Histological and ultra-structural studies of cardiac tissues supported the protective characteristics of SR. CONCLUSIONS: From the present findings it can be concluded that, SR could offer protection against T2DM and its associated cardio-toxicity via multiple mechanisms viz. hypoglycemic, antioxidant and anti-inflammatory actions.

Effect of Acacia catechu (L.f.) Willd. on Oxidative Stress with Possible Implications in Alleviating Selected Cognitive Disorders.

In human body, several categories of degenerative processes are largely determined by free radicals originating in cell. Free radicals are also known to have correlated with a variety of cognitive disorders (CDs) resulting in neuronal injury and eventually to death. Alzheimer's disease (AD) and Parkinson's disease (PD) are such kind of killer CDs that occur due to dysfunction of cholinergic and dopaminergic neurons. Plant parts of Ginkgo biloba, Bacopa monnieri etc. are being used for the treatment of cognitive disorders in several countries. The present study was aimed to explore the detailed antioxidant and anti-cholinesterase activity of Acaciacatechu leaf (ACL) over CDs. Gas chromatography-Mass spectroscopy (GC-MS) analysis and Nuclear Magnetic Resonance (NMR) were employed to identify the bioactive components present in ACL. Furthermore, the extract was evaluated to check the cytotoxic effects of ACL on normal cells. Amongst several antioxidant assays, DPPH assay, hydroxyl radical, nitric oxide radical and hypochlorous acid inhibitory activities were found to be greater in ACL than that of the respective standards while other assays exhibited a moderate or at per inhibitory activity with standards. Total phenolic and flavonoid content were also found to be present in decent amount. In addition, we found, a greater acetylcholinesterase (AChE) inhibitory activity of ACL when compared to other medicinally important plants, indicating its positive effect over CDs. Forty one bioactive components were explored through GC-MS. Of these, gallic acid, epicatechin, catechin, isoquercitrin etc. were found, which are potent antioxidant and a few of them have anti-neurodegenerative properties. Eventually, ACL was found to be nontoxic and safer to consume. Further studies with animal or human model however, would determine its efficacy as a potential anti-schizophrenic drug.

Metabolic diversity in the grains of Indian varieties of rice.

The aim of the present work was to analyze metabolic diversity in 26 different indica varieties of rice grains. Seventy-six metabolites could be identified in the methanol extracts of each of the rice varieties analyzed by gas chromatography-mass spectrometry. These metabolites included 9 sugars/sugar alcohols, 17 amino acids/derivatives, 18 fatty acids, 5 free phenolic acids and 19 other organic acids, 3 phytosterols, 5 other constituents. Cluster analyses to extract information for similarity and differences in metabolites unveiled diversity in metabolite profile. Two hierarchical clusters were generated based on the metabolite contents of the rice varieties. The first cluster (cluster I) consisted of one variety only. The second cluster again segregated into four clusters (clusters II, III, IV and V). Very distinct differences were visible amongst the clusters with respect to their sugars/sugar alcohols, organic acid, amino acid and fatty acid, phenol, and sterol profiles. Metabolites determine nutritional quality, taste, aroma. This and future efforts on the metabolomic information would help biochemists and nutritionists to better understand the nutritional quality of such grains at varietal level and correlating metabolites and long term human health related issues.

Elicitation of diosgenin production in Dioscorea floribunda by ethylene-generating agent.

Dioscorea floribunda cells aggregates were cultured in liquid modified MS medium supplemented with 2,4-D (2 mg/l) and kinetine (0.1 mg/l). The cells were treated with different concentrations of ethylene-generating-agent 2-chloroethylphosphonic acid (2-CEPA). 2-CEPA at concentrations of 50 mg/l, 100 mg/l elicited production of diosgenin. The level of diosgenin increased to 72-fold higher than those of non-treated cells with 100 mg/l of 2-CEPA.

Steroidal compounds from in vitro regenerated shoots of Datura metel.

Regeneration of adventitious shoots from young leaves of Datura metel is described. Shoot buds developed on MS medium with 2 mg/l and elongated on hormone-free solid basal medium. The microshoots failed to produce alkaloids, but a number of steroidal compounds were detected. The C(28) sterol 3beta,24xi-dihydroxy-ergosta-5,25-dienolide, regarded as the precursor of withanolides and related steroidal lactones, was identified. The withanolide 12-deoxywithastramonolide was also detected.