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Multidrug and toxic compound extrusion (MATE) is a family of membrane transporters that transport ions and secondary metabolites like anthocyanins in plants. Although various research on MATE has been carried out on different plant species, this is the first comprehensive report to exploit the Daucus carota genome to identify the MATE gene family. We investigated the chromosomal distribution, gene duplication, motif identification, cis-element analysis, in vitro cultures, stress treatment, protein docking, and simulation of DcMATEs. Our studies identified 45 DcMATEs through genome-wide analysis and detected five segmental and six tandem duplications from its genome. The chromosome distribution, phylogenetic analysis, and cis-regulatory elements revealed the structural diversity and numerous functions associated with the DcMATEs. Further, we quantified anthocyanin content and identified the co-expression of DcMATE21 with anthocyanin biosynthesis genes in in vitro cultures under ABA, SA, MJ, SNP, and PHE stress treatments. In addition, membrane dynamics of DcMATE21 with anthocyanin (cyanidin-3-glucoside) identified the binding pocket showing extensive H-bond interactions with ten amino acids present in the transmembrane helix of 7, 8, and 10 of DcMATE21. This investigation speculates the involvement of DcMATE21 in anthocyanin accumulation in in vitro cultures of D. carota.

Serotonin and melatonin are important signaling and stress mitigating molecules. However, their role and molecular mechanism in the accumulation of isoflavones are not clearly defined. To elucidate...

Folate (vitamin B

Serotonin (SER) and melatonin (MEL) are signalling molecules involved in signalling in various stress response mechanisms in plants. However, their role and molecular mechanism in the accumulation of isoflavones and biomass under temperature stress are not clearly defined. To explicate their function, SER and MEL at different ratios were applied to the in vitro cultures of soybean at 16, 24, and 32 °C and were analyzed for the accumulation of bioactive compounds, isoflavones content, and culture biomass followed by transcript levels of cell division, isoflavones, and ethylene biosynthesis genes. An increase in biomass, total phenolics, total flavonoids, and different forms of isoflavones content was evident in the treatments. The expression level of critical genes of isoflavone (chalcone synthase, chalcone reductase, chalcone isomerase, and isoflavone synthase), ethylene (s-adenosyl methionine and 1-aminocyclopropane-1-carboxylic acid oxidase), cell division (cyclin and cyclin-dependent kinase), and MEL biosynthesis genes (acetyl serotonin O-methyltransferase and serotonin N-acetyltransferase) and transcription factors (melatonin 2 hydroxylase, myeloblastosis like protein) were co-expressed during temperature stress in the presence of SER and MEL. At 32 °C, the SER and MEL application showed maximum biomass and isoflavones content and was substantiated with the corresponding gene expression. We hypothesize that the SER and MEL ratio may play a role in increasing the biomass and isoflavones content under temperature stress in soybean cell culture.

Serotonin and melatonin are important signaling and stress mitigating molecules. However, their role and molecular mechanism in the accumulation of isoflavones are not clearly defined. To elucidate their functions, serotonin and melatonin were applied to

The present study investigated the potential of a small molecule inhibitor, 1-deoxynojirimycin (DNJ), to extend the shelf life of tomatoes. The optimum concentration of DNJ and the proper ripening stage for treatment were standardized using response surface methodology, following a central composite design. The concentration of DNJ used for the analysis was 0.15 mM, and 0.30 mM and the ripening stages of the tomato fruit analysed were immature green, mature green, breaker, ripen and over-ripen. Analysis of the influence of the DNJ treatment of the fruit using quadratic multiple regression models considering the factors colour, texture, and free sugars revealed significant responses. A DNJ concentration of 0.30 mM and fruit-ripening stage of mature green was found to be optimal for the treatment. DNJ-treatment maintained fruit firmness throughout ripening with a significant reduction in reducing sugar formation. Enzyme activity of the N-glycan processing enzymes involved in cell wall softening, α-mannosidase and β-d-N-acetylhexosaminidase revealed a significant reduction in their activity by 2 and 3.5-fold, respectively. Down-regulation of expression of important ripening-related and softening process-associated genes, aminocyclopropane carboxylic synthase-4, aminocyclopropane carboxylic oxidase, polygalacturonase and pectin methylesterases at 4, 5, 6 and 5-fold, respectively, was also observed. The present results showed that the treatment of mature green tomato fruit with DNJ at a concentration of 0.30 mM can delay the ripening of the tomato fruit by inhibiting cell wall and N-glycan processing enzymes.

The present study investigated the potential of a small molecule inhibitor, 1-deoxynojirimycin (DNJ), to extend the shelf life of tomatoes. The optimum concentration of DNJ and the proper ripening stage for treatment were standardized using response surface methodology, following a central composite design. The concentration of DNJ used for the analysis was 0.15 mM, and 0.30 mM and the ripening stages of the tomato fruit analysed were immature green, mature green, breaker, ripen and over-ripen. Analysis of the influence of the DNJ treatment of the fruit using quadratic multiple regression models considering the factors colour, texture, and free sugars revealed significant responses. A DNJ concentration of 0.30 mM and fruit-ripening stage of mature green was found to be optimal for the treatment. DNJ-treatment maintained fruit firmness throughout ripening with a significant reduction in reducing sugar formation. Enzyme activity of the N-glycan processing enzymes involved in cell wall softening, α-mannosidase and β-d-N-acetylhexosaminidase revealed a significant reduction in their activity by 2 and 3.5-fold, respectively. Down-regulation of expression of important ripening-related and softening process-associated genes, aminocyclopropane carboxylic synthase-4, aminocyclopropane carboxylic oxidase, polygalacturonase and pectin methylesterases at 4, 5, 6 and 5-fold, respectively, was also observed. The present results showed that the treatment of mature green tomato fruit with DNJ at a concentration of 0.30 mM can delay the ripening of the tomato fruit by inhibiting cell wall and N-glycan processing enzymes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-020-02196-3) contains supplementary material, which is available to authorized users.

The interaction between pentoxifylline (PTX) and bovine serum albumin (BSA) was studied under physiological condition by spectroscopic, voltammetry and molecular modelling techniques. The results of fluorescence studies revealed that the quenching mechanism of BSA by PTX was a static procedure. Binding constant of PTX-BSA was calculated and its value found to be 8.895 × 104 M−1, which is in close agreement with the results obtained from UV-Visible and voltammetry approach. The negative values of thermodynamic parameters (ΔG, ΔH and ΔS) indicated that van der Waals force and hydrogen bonding play major roles in the interaction of PTX with BSA. Based on Forster’s theory of non-radiation energy transfer, the binding distance (r) between the donor (BSA) and acceptor (PTX) was found to be 5.38 nm (298 K). The results of UV-Visible and circular dichroism (CD) spectroscopy showed that PTX interacts with BSA and induces conformational changes by reducing the α-helix content. The results of the UV-visible and voltammetry studies were further verified by the molecular docking method. Molecular modelling studies revealed possible residues involved in the drug-protein interaction and indicated that PTX binds to Site IIA of BSA.

The interaction of covellite hexagonal phase of copper sulphide nanoparticles (CuS NPs) with bovine serum albumin (BSA) was examined systematically by using fluorescence, UV–visible, circular dichroism (CD), Fourier transform infrared (FTIR), dynamic light scattering (DLS) and molecular modelling techniques. Electrochemical method was studied to further confirm the interaction of BSA with CuS NPs. The results of fluorescence studies demonstrated that fluorescence of BSA was quenched by CuS NPs via a static quenching mechanism. The negative values of thermodynamic parameters (ΔG, ΔH and ΔS) indicated that the binding process is spontaneous, exothermic and van der Waals force or hydrogen bonding plays major roles in the interaction of CuS NPs with BSA. The interaction of CuS NPs with Trp residue was established by synchronous studies, and competitive binding studies revealed that Trp-212 of subdomain IIA was involved in the interaction with these nanoparticles. Further, the efficiency of energy transferred and the distance between fluorophore (BSA) and acceptor (CuS NPs) were calculated using Forster’s resonance energy transfer theory. The results of UV–visible, CD, FTIR and DLS revealed that the CuS NPs interact with BSA by inducing the conformational changes in secondary structure and reducing the α-helix content of BSA. Molecular modelling studies suggested that CuS NPs bind to site I of sub domain IIA of BSA. The results of spectroscopic and molecular docking studies were complimented by the electrochemical techniques.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease. The pathogenesis of interstitial lung diseases, including its most common form, IPF, remains poorly understood. Alveolar epithelial cell (AEC) apoptosis, proliferation, and accumulation of myofibroblasts and extracellular matrix deposition results in progressive loss of lung function in IPF. We found induction of tumor suppressor protein, p53, and apoptosis with suppression of urokinase-type plasminogen activator (uPA) and the uPA receptor in AECs from the lungs of IPF patients, and in mice with bleomycin, cigarette smoke, silica, or sepsis-induced lung injury. Treatment with the caveolin-1 scaffolding domain peptide (CSP) reversed these effects. Consistent with induction of p53, AECs from IPF lungs or mice with diverse types of lung injuries showed increased p53 acetylation and miR-34a expression with reduction in Sirt1. This was significantly reduced after treatment of wild-type mice with CSP, and uPA-deficient mice were unresponsive. Bleomycin failed to induce miR-34a in p53- or plasminogen activator inhibitor-1 (PAI-1)-deficient mice. CSP-mediated inhibition of miR-34a restored Sirt1, suppressed p53 acetylation and apoptosis in injured AECs, and prevented pulmonary fibrosis (PF). AEC-specific suppression of miR-34a inhibited bleomycin-induced p53, PAI-1, and apoptosis and prevented PF, whereas overexpression of precursor-miR-34a increased p53, PAI-1, and apoptosis in AECs of mice unexposed to bleomycin. Our study validates p53–miR-34a feedback as a potential therapeutic target in PF.

Nitric oxide (NO) is known to antagonize ethylene by various mechanisms, one of such mechanisms is reducing ethylene levels by competitive action on S-adenosyl-L-methionine (SAM)&mdash, a common precursor for both ethylene and polyamines (PAs) biosynthesis. In order to investigate whether this mechanism of SAM pool diversion by NO occur towards PAs biosynthesis in banana, we studied the effect of NO on alterations in the levels of PAs, which in turn modulate ethylene levels during ripening. In response to NO donor sodium nitroprusside (SNP) treatment, all three major PAs viz. putrescine, spermidine and spermine were induced in control as well as ethylene pre-treated banana fruits. However, the gene expression studies in two popular banana varieties of diverse genomes, Nanjanagudu rasabale (NR, AAB genome) and Cavendish (CAV, AAA genome) revealed the downregulation of SAM decarboxylase, an intermediate gene involved in ethylene and PA pathway after the fifth day of NO donor SNP treatment, suggesting that ethylene and PA pathways do not compete for SAM. Interestingly, arginine decarboxylase belonging to arginine-mediated route of PA biosynthesis was upregulated several folds in response to the SNP treatment. These observations revealed that NO induces PAs via l-arginine-mediated route and not via diversion of SAM pool.

In view of exploring the potential of pyrimidine derivatives as anticancer agents, a series of 4-methyl-6-morpholinopyrimidine derivatives was synthesised and characterised by NMR (H-1 & C-13), SC-XRD and mass spectral analysis. The in vitro anticancer activity of these compounds was investigated using different human cancer cell lines, namely HeLa (cervix), NCI-H460 (lung), MCF-7 (breast), HepG2 (liver) and IMR-32 (brain). Compounds 4c and 5h exhibited potent anticancer activity in a dose-dependent manner as compared to other derivatives, with IC50 values of 5.88 +/- 1.22 and 6.11 +/- 2.12 mu M on HeLa and NCI-H460, cells respectively. The inhibitory effect of 4c and 5h on cancer cell proliferation was shown to be a consequence of reactive oxygen species (ROS) generation and subsequent induction of cellular apoptosis, as evidenced by an increase in hypodiploid (subG1) population, early apoptotic cell population, caspase-3/7 activity, loss of mitochondrial membrane potential and degradation of nuclear DNA. Furthermore, molecular docking studies revealed that 4c and 5h compounds bind to the ATP binding pocket of the mammalian target of rapamycin (mTOR). Based on our results, we conclude that 4-methyl-6-morpholinopyrimidine derivatives prevent cancer cell proliferation by inducing apoptosis and thus have potential to be further explored for anticancer properties.

Increasing the quantity of natural folates in plant foods is recently gaining significant interest, owing to their acute deficiencies in various populations. This study observed that foliar salicylic acid treatment enhanced the accumulation of folates in Arabidopsis, which correlated with the increase in a folate binding protein (FBP) and the expression of mRNA of a putative folate binding protein At5G27830. A protein band corresponding to ∼43 kDa was observed after resolving the affinity-purified protein on SDS-PAGE, and the partial amino acid sequence indicated that the protein is indeed At5G27830. Docking studies performed with At5G27830 confirmed specific binding of folic acid to predicted site. Heterologous expression of At5G27830 in the yeast resulted in significant uptake and accumulation of folic acid in cells. This novel study of a plant FBP will be useful for folate metabolic engineering of a wide range of crops.

Coriander (Coriandrum sativum L.), a commonly used annual herb that accumulates carotenoids upon methyl jasmonate (MeJA) treatment, provides an excellent model to investigate carotenogenesis and gene regulation. To explore key mechanisms involved in enhancing carotenoids, transcriptional expression profile of ten carotenogenic genes in the presence of MeJA and various gene specific inhibitors were investigated. Foliar application of MeJA (10 μM) increased expression levels of CsPDS (phytoene desaturase), CsZDS (ς-carotene desaturase), CsCHYE (carotene e - hydroxylase) and CsLCYE (lycopene β-cyclase) genes, and their transcript levels were strongly associated with carotenoid content, where, three days after treatment, 3.9 & 6.1 fold increase was observed for β-carotene and lutein respectively. The regulatory effect of key genes, CsPDS, CsZDS, CsLCYE and LCYB were further confirmed by using gene-specific inhibitors fosmidomycin, norflurazon and amitrol. Norflurazon- the phytoene desaturation inhibitor leads to a decrease in β-carotene and lutein content correlated with CsPDS, CsZDS gene induction. Our results clearly demonstrate that MeJA induced-signalling network evokes carotenogenic genes, leading to the accumulation of carotenoids. This knowledge may help to develop precise strategies for remodelling carotenoid pathway so that desired levels of a particular carotenoid in leafy vegetables is achievable.

Pulmonary surfactant protein C (SP-C) expression by type II alveolar epithelial cells (AECs) is markedly reduced in diverse types of lung injuries and is often associated with AEC apoptosis. It is unclear whether loss of SP-C contributes to the increased p53 and urokinase-type plasminogen activator (uPA) system cross-talk and apoptosis of AECs. Therefore, we inhibited SP-C expression in human and murine AECs using lentivirus vector expressing shRNA and tested p53 and downstream changes in the uPA-fibrinolytic system. Inhibition of SP-C expression in AECs induced p53 and activated caspase-3, indicating AEC apoptosis. We also found that bleomycin or cigarette smoke exposure failed to inhibit SP-C expression or apoptosis in AECs in p53- and plasminogen activator inhibitor-1 (PAI-1)-deficient mice. Depletion of SP-C expression by lentiviral SP-C shRNA in PAI-1-deficient mice failed to induce p53 or apoptosis in AECs, whereas it increased both AEC p53 and apoptosis in wild-type and uPA-deficient mice. SP-C inhibition in AECs also increased in CXCL1 and CXCL2 and their receptor CXCR2 as well as ICAM-1 expression, which is indicative of a proinflammatory response. Overexpression of p53-binding 3′-UTR sequences in AECs inhibited PAI-1 induction while maintaining uPA and uPAR protein and mRNA expression. Furthermore, caveolin-1 expression and phosphorylation were increased in AECs, indicating an intricate link between caveolin-1 and Src kinase-mediated cell signaling and AEC apoptosis due to loss of SP-C expression through p53 and uPA system-mediated cross-talk. The role of uPA, PAI-1, and p53 in the regulation of AEC apoptosis after injury was also determined in knockout mice.

The present study reports the presence of folate binding proteins (FBPs) in the plants, coriander and Arabidopsis, and their contributions toward folate enhancement. After observing that salicylic acid (SA) enhanced the accumulation of folates in coriander, a study was conducted in Arabidopsis, where twofold increase in folates occurred in foliage upon SA treatment. For obtaining insights into genes involved in SA-induced folate accumulation, microarray data of responsive genes in Arabidopsis were screened. Based on the expression profiles, 19 genes were further analysed by qPCR. The results revealed that folate biosynthetic genes were largely down-regulated, whereas a gene of a putative folate-binding protein (FPB) was up-regulated, which correlated with a significant increase of FPBs in foliage. This new information on a plant FBP appears useful for metabolic engineering of a wide range of crops to enhance the content and stability of the folates during post-harvest storage.

Foliage of coriander crop is a rich source of pro-vitamin-A carotenoids and phenolics of relevance to human health. Foliar-application of plant growth regulators e methyl jasmonate and salicylic acid, differentially elicited total carotenoids, b-carotene, lutein, chlorophylls, total phenolics and chlorogenic acid in coriander, GS4 Multicut and Mahak, the cultivars with high and low carotenoids respectively. Carotenoids and total phenolics increased 6.8 and 3 folds respectively when treated with methyl jasmonate (10 mmol/L), whereas salicylic acid (500 mmol/L) showed 5.4 and 3.5 folds of respective compounds. These treatments also enhanced levels of b-carotene, lutein, chlorophylls and chlorogenic acid, as observed by HPLC/MS analyses. Carotenoid and phenolic extracts of the best treatments showed significant increase in hydroxyl and superoxide radical scavenging antioxidant activities compared with controls. The observations made here indicate that the precise use of plant growth regulators is a simple method for naturally augmenting the nutritionally important compounds in coriander crop.

Since vitamin A deficiency is prevalent in many developing countries, we sought to focus research on local, affordable and well-accepted sources of provitamin A carotenoids. As dessert bananas are consumed fresh round the year and processed as products, this study investigated whether postclimacteric biochemical changes are linked to carotenoid degradation in four Indian varieties, one commercial (Cavendish, AAA), one Red banana (genome AAA) and two locally-important ones (genome AAB). Despite large differences in their ripening characteristics, textural loss was lesser in AABs than AAAs. High levels of carotenoids (21.0 mg/g FW), b-carotene (9.14 mg/g FW) and a-carotene (9.32 mg/g FW) were found in Red banana accounting for retinol activity equivalent of 114 mg/100 g FW. The carotenoid levels were lower in two local varieties and lowest in Cavendish, with no post-climacteric loss. Dry grits, prepared using Red banana pulp, milk powder and sugar, retained about 70% carotenoids and appeared useful in confectionaries.

Although populations of several tropical countries are under severe vitamin-A deficiency, traditional sources of pro-vitamin-A carotenoids have not been exploited due to the lack of relevant research data. In this study, ten commercial varieties of coriander (Coriandrum sativum L.), grown at identical conditions, were evaluated for carotenoids, their bio-efficacy and stability during drying, with the main emphasis on β-carotene (vitamin-A precursor) analysis by HPLC-MS. In all varieties, β-carotene content was higher in foliage at mature stage, than in seedlings and seeds. Variety GS4 Multicut produced highest biomass (6.18 ± 0.73 g/plant), total carotenoids (217.50 ± 5.6 mg/100 g DW) and β-carotene (73.64 ± 0.3 mg/100 g DW) at pre-flowering stage. Carotenoids extract showed a high antioxidant activity with IC50 value of 14.29 ± 1.68 μg/ml, scavenging hydroxyl radicals and rendering higher protection to DNA than by standard gallic acid (IC50 = 357.21 ± 4.29). Microwave drying of foliage was rapid with better retention of pigments, high intactness of trans-β-carotene and higher extractability of pigments when compared with oven drying.

Folate (vitamin B9) content was evaluated in 10 varieties of coriander with the aim of enhancing its concentration and stability, because of three reasons: 1) coriander is among a few widely used greens in the world and suits many cuisines, 2) folate deficiency is prevalent in developing countries causing anaemia, infant mortality and neural tube closure defects, and 3) natural folate is preferred due to doubts about health risks associated with the synthetic form. In C. sativum, the highest folate content of 1,577 μg/100 g DW was found in var. GS4 Multicut foliage of mature plants (marketable stage) with an insignificantly higher content (1,599.74 μg/100 g DW) at flowering, which is a stage not preferred in markets. In callus cultures treated with plant growth regulators (GRs) (6-benzylaminopurine, kinetin and abscisic acid) substantial increase in folate occurred after 6 h, whereas elicitors (methyl jasmonate and salicylic acid) caused rapid 2-fold increase of folate, particularly in response to salicylic acid. Based on these observations, foliar applications were done for in vivo plants, where salicylic acid (250 μM, 24 h) also enhanced folate level by 2-folds (3,112.33 μg/100 g DW), although the content varied with diurnal rhythms. Stability of folates in treated coriander foliage was 10 % higher than in untreated foliage when stored at 25 °C and 4 °C. This study has established for the first time that coriander foliage is rich in folates, which can be doubled by elicitation and impart 10 % more stability than control during processing and storage.