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Heavy metals are ubiquitously present in nature, including soil, water, and thus in plants, thereby causing a potential health risk. This study has investigated the role and efficiency of the chickpea metallothionein 1 (MT1) gene against the major toxic heavy metals, i.e., As [As(III) and As(V)], Cr(VI), and Cd toxicity. MT1 over-expressing transgenic lines had reduced As(V) and Cr(VI) accumulation, whereas Cd accumulation was enhanced in the L3 line. The physiological responses (WUE, A, Gs, E, ETR, and qP) were noted to be enhanced in transgenic plants, whereas qN was decreased. Similarly, the antioxidant molecules and enzymatic activities (GSH/GSSG, Asc/DHA, APX, GPX, and GRX) were higher in the transgenic plants. The activity of antioxidant enzymes, i.e., SOD, APX, GPX, and POD, were highest in the Cd-treated lines, whereas higher CAT activity was observed in As(V)-L1 and GRX in Cr-L3 line. The stress markers TBARS, H2O2, and electrolyte leakage were lower in transgenic lines in comparison to WT, while RWC was enhanced in the transgenic lines, and the transcript of MT1 gene was accumulated in the transgenic lines. Similarly, the level of stress-responsive amino acid cysteine was higher in transgenic plants as compared to WT plants. Among all the heavy metals, MT1 over-expressing lines showed a highly increased accumulation of Cd, whereas a non-significant effect was observed with As(III) treatment. Overall, the results demonstrate that Arabidopsis thaliana transformed with the MT1 gene mitigates heavy metal stress by regulating the defense mechanisms in plants.

Anaerobic ammonium oxidation (anammox) is a bacterial process in which ammonium and nitrite are combined into dinitrogen gas and water, yielding energy for the cell. This process relies on a series of redox reactions catalyzed by a set of enzymes, with electrons being shuttled to and from these enzymes, likely by small cytochrome c proteins. For this system to work productively, these electron carriers require a degree of specificity toward the various possible redox partners they encounter in the cell. Here, we compare two cytochrome c proteins from the anammox model organism Kuenenia stuttgartiensis. We show that they are highly homologous, are expressed at comparable levels, share the same fold, and display highly similar redox potentials, yet one of them accepts electrons from the metabolic enzyme hydroxylamine oxidase (HAO) efficiently, whereas the other does not. An analysis of the crystal structures supplemented by Monte Carlo simulations of the transient redox interactions suggests that this difference is at least partly due to the electrostatic field surrounding the proteins, illustrating one way in which the electron carriers in anammox could attain the required specificity. Moreover, the simulations suggest a different “outlet” for electrons on HAO than has traditionally been assumed.

Glutaredoxins (Grxs) are short, cysteine-rich glutathione (GSH)-mediated oxidoreductases. In this study, a chickpea (Cicer arietinum L.) glutaredoxin [LOC101493651 (CaGrx)] gene has been selected based on screening experiments with two contrasting varieties of chickpea, PUSA-362 (drought-tolerant) and ICC-1882 (drought-sensitive) under drought and salinity. The tolerant variety showed higher CaGrx gene expression, as compared to less in the sensitive variety, under both the stresses. The CaGrx gene was then over-expressed in Arabidopsis thaliana and were exposed to drought and salinity. The over-expression of CaGrx elevated the activity of glutaredoxin, which induced antioxidant enzymes (glutathione reductase; GR, glutathione peroxidase; GPX, catalase; CAT, ascorbate peroxidase; APX, glutathione-S-transferase; GST, superoxide dismutase; SOD, monodehydroascorbate reductase; MDHAR, and dehydroascorbate reductase; DHAR), antioxidants (GSH and ascorbate) and stress-responsive amino acids (cysteine and proline). Enhancement in the antioxidant defense system possibly administered tolerance in transgenics against both stresses. CaGrx reduced stress markers (H(2)O(2), TBARS, and electrolyte leakage) and enhanced root growth, seed germination, and survival against both stresses. The physiological parameters (net photosynthesis; P(N), water use efficiency; WUE, stomatal conductance; g(s), transpiration; E, electron transport rate; ETR, and photochemical quenching; qP), chlorophylls and carotenoids, were improved in the transgenics during both stresses, that maintained the photosynthetic apparatus and protected the plants from damage. The enhanced activity of the cysteine biosynthesis enzyme, o-acetylserine (thiol) lyase (OAS-TL), increased the cysteine level in the transgenics, which elevated glutathione biosynthesis to maintain the ascorbate–glutathione cycle under both stresses. This investigation verified that the CaGrx gene provides tolerance against salinity and drought, maintaining physiological and morphological performances, and could be exploited for genetic engineering approaches to overcome both the stresses in various crops. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-00999-z.

Rhizobacterial diversity is an indicator of soil health and in turn it is influenced by the host crops, edaphic factors and weather. The present investigation reports the diversity of endospore forming, gram positive rhizobacteria (Bacillus megaterium, B. thuringiensis, B. cereus and Lysinibacillus spp.) inhabiting 11 different agricultural crops in the IGP of India. Twelve Bacillus thuringensis (Bt) isolates were identified by screening 62 g positive bacterial isolates from 63 rhizosperic soil samples collected from 6 districts of IGP. The highest Bt index was recorded from Bhopal (0.25) followed by Fatehpur (0.17), Kanpur dehat (0.10), Jalaun (0.08) and Hamirpur (0.06). The bacterial isolates were characterized based on the 16SrRNA gene and phylogenetically grouped into 3 clades. The spore crystal mixture of 12 Bt isolates were subjected to insect bioassay and it revealed, F8.IIPR has highest toxicity against Spilosoma obliqua Walker (100%), Olepa ricini Fabricius (91.67%) and Helicoverpa armigera Hubner (100%) larva. Survival analysis (ST50) showed that the S. obliqua is highly susceptible than O. ricini and H. armigera to F8.IIPR. Crystal staining and protein profiling showed the presence of cry 1 (135 kDa) and cry 2 (65 kDa) genes in 5 Bt isolates. PCR amplification of vip3A gene confirmed its presence in five Bt isolates. To conclude F8.IIPR and Ak2.IIPR has the potential as a promising biopesticide for controlling the three lepidopteran insects tested.

Cytoplasmic male sterility(CMS), a maternally inherited trait, provides a promising means to harness yield gains associated with hybrid vigor. In pigeonpea [Cajanus cajan (L.) Huth], nine types of sterility‐inducing cytoplasm have been reported, of which A2 and A4 have been successfully deployed in hybrid breeding. Unfortunately, molecular mechanism of the CMS trait is poorly understood because of limited research invested. More recently, an association between a mitochondrial gene (nad7) and A4‐CMS has been demonstrated in pigeonpea; however, the mechanism underlying A2‐CMS still remains obscure. The current investigation aimed to analyze the differences in A2‐CMS line (ICPL 88039A) and its isogenic maintainer line (ICPL 88039B) at transcriptome level using next‐generation sequencing. Gene expression profiling uncovered a set of 505 genes that showed altered expression in response to CMS, of which, 412 genes were upregulated while 93 were downregulated in the fertile maintainer line vs. the CMS line. Further, gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein–protein interaction (PPI) network analyses revealed association of CMS in pigeonpea with four major pathways: glucose and lipid metabolism, ATP production, pollen development and pollen tube growth, and reactive oxygen species (ROS) scavenging. Patterns of digital gene expression were confirmed by quantitative real‐time polymerase chain reaction (qRT‐PCR) of six candidate genes. This study elucidates candidate genes and metabolic pathways having potential associations with pollen development and male sterility in pigeonpea A2‐CMS. New insights on molecular mechanism of CMS trait in pigeonpea will be helpful to accelerate heterosis utilization for enhancing productivity gains in pigeonpea.

Contains fulltext : 209065.pdf (Publisher’s version ) (Open Access) Anaerobic ammonium oxidation (anammox) is a microbial process responsible for significant nitrogen loss from the oceans and other ecosystems. The redox reactions at the heart of anammox are catalyzed by large multiheme enzyme complexes that rely on small cytochrome c proteins for electron shuttling. Among the most highly abundant of these cytochromes is a unique heterodimeric complex composed of class I and class II c-type cytochrome called NaxLS, which has distinctive biochemical and spectroscopic properties. Here, we present the 1.7 Å resolution crystal structure of this complex from the anammox organism Kuenenia stuttgartiensis (KsNaxLS). The structure reveals that the heme irons in each subunit exhibit a rare His/Cys ligation, which, as we show by substitution, causes the observed unusual spectral properties. Unlike its individual subunits, the KsNaxLS complex binds nitric oxide (NO) only at the distal heme side, forming 6cNO adducts. This is likely due to steric immobilization of the proximal heme binding motifs upon complex formation, a finding that may be of functional relevance, since NO is an intermediate in the central anammox metabolism. Pulldown experiments with K. stuttgartiensis cell-free extract showed that the KsNaxLS complex binds specifically to one of the central anammox enzyme complexes, hydrazine synthase, which uses NO as one of its substrates. It is therefore possible that the KsNaxLS complex plays a role in binding the volatile NO to retain it in the cell for transfer to hydrazine synthase. Alternatively, we propose that KsNaxLS may shuttle electrons to this enzyme complex. 22 september 2019

Purpose The purpose of this paper is to extract, characterise and quantify curcumin from selected Zingiberaceae of “kunyit” or turmeric (Curcuma longa), “temu lawak” or Javanese turmeric (Curcuma xanthorrhiza), “temu pauh” (Curcuma mangga), “lempoyang” (Zingiber zerumbet) and “bonglai” (Zingiber cassumunar) using alkaline and chemical-based extraction method for antimicrobial and antioxidant activities. Design/methodology/approach Through the alkaline-based extraction method, all parts of rhizome samples were freeze-dried for 72 h before grounded into a fine powder and kept at −20°C. The powdered sample (0.1 g) was weighed and placed in a 50 mL tube. About 20 mL of 2 M NaOH solution was added into the tube. The solution was allowed to stand for 30 min. Then, 20 mL of ethyl acetate was added into the tube. The solution was mixed well then centrifuged at 13,500 rpm for 3 min. The upper layer was collected using a pipette. The process was repeated until the upper layer became almost colourless. The collected ethyl acetate solution was concentrated using a rotary evaporator to remove the ethyl acetate from the extracted compound. The concentrated curcumin was placed in a universal bottle, which was then dried from the remaining ethyl acetate using nitrogen drying process. The dried curcumin was then stored inside the freezer at −20ºC. The antimicrobial activities were using agar diffusion method against bacterial and fungi, while the antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay. Findings All the samples successfully showed a single peak (curcumin) that gained from the high-performance liquid chromatography (HPLC) chromatogram analysis (at 425 nm) using the alkaline-based extraction method and the highest curcumin content was in turmeric (12.95 ± 1.07mg/g DW). At 10.0 mg/mL curcumin concentration, the best antibacterial activity was against on methicillin-resistant staphylococcus aureus (MRSA) with 7.50 ± 0.71 mm inhibition zone, while the best antifungal activity was against on Aspergillus niger with 8.00 ± 0.41 mm inhibition zone. The DPPH antioxidant test resulted in the highest inhibition (110.41 per cent) was at 0.25 mg/mL curcumin concentration. Originality/value Through HPLC analysis, all samples successfully showed a single peak of curcumin at 425 nm. The total carotenoid determination from turmeric revealed that the samples content was substantially higher using alkaline-based extraction (18.40 ± 0.07 mg/g DW) compared to chemical-based extraction (9.42 ± 0.20 mg/g ± SD).

A multi-technique approach has been used to probe the interaction of model protein bovine serum albumin (BSA) with a series of substantially biodegradable and less cytotoxic ethylene-oxide linked ester-functionalized gemini surfactants (designated as Cm-E2O-Cm, m = 12, 14 and 16). Binding of BSA with the gemini surfactants has been deduced on the basis of steady state fluorescence measurements. Synchronous fluorescence spectra indicate an exposure of the tyrosine residues and entrapment of the tryptophan residues into the hydrophobic cloud of the gemini surfactants. Post surfactant addition, pyrene fluorescence studies depict mild fluctuations in the micropolarity experienced by the probe molecules encapsulated in the protein. Temperature variation fluorescence and UV absorption studies confirm ground state complexation between BSA and the concerned gemini surfactants. Far-UV CD spectra are indicative of slight uncoiling of the alpha helices. These experimental findings are corroborated by the molecular docking results which predict that the probable binding sites for Cm-E2O-Cm gemini surfactants are in the vicinity of aromatic residues. Thus, the investigations presented herein elucidate the binding of serum albumin with surfactants of optimized architecture. Therefore, with further exhaustive screening tests, the Cm-E2O-Cm gemini surfactants may be potentially utilized in pharmaceuticals, cosmetics and other broad application fields.

Anaerobic Ammonium Oxidation ("anammox") is a bacterial process in which nitrite and ammonium are converted into nitrogen gas and water, yielding energy for the cell. Anammox is an important branch of the global biological nitrogen cycle, being responsible for up to 50% of the yearly nitrogen removal from the oceans. Strikingly, the anammox process uniquely relies on the extremely reactive and toxic compound hydrazine as a free intermediate. Given its global importance and biochemical novelty, there is considerable interest in the enzymes at the heart of the anammox pathway. Unfortunately, obtaining these enzymes in sufficiently large amounts for biochemical and structural studies is problematic, given the slow growth of pure cultures of anammox bacteria when high cell densities are required. However, the anammox process is being applied in wastewater treatment to remove nitrogenous waste in processes like DEamMONification (DEMON). In plants using such processes, which rely on a combination of aerobic ammonia-oxidizers and anammox organisms, kilogram amounts of anammox bacteria-containing sludge are readily available. Here, we report a protein isolation protocol starting from anammox cells present in DEMON sludge from a wastewater treatment plan that readily yields pure preparations of key anammox proteins in the tens of milligrams, including hydrazine synthase HZS and hydrazine dehydrogenase (HDH), as well as hydroxylamine oxidoreductase (HAO). HDH and HAO were active and of sufficient quality for biochemical studies and for HAO, the crystal structure could be determined. The method presented here provides a viable way to obtain materials for the study of proteins not only from the central anammox metabolism but also for the study of other exciting aspects of anammox bacteria, such as for example, their unusual ladderane lipids.

The fluoroquinolone antibiotic drug namely ciprofloxacin hydrochloride (CFH) is widely prescribed for the treatment of different bacterial infections. The interaction of CFH with a synthetic polymer, polyvinyl pyrrolidone (PVP), and biopolymer, bovine serum albumin (BSA) was studied by UVvisible and fluorescence spectroscopic methods at different temperatures. The binding constant (K b ) for the CFH-PVP complex was determined from the Benesi-Hildebrand plot. PVP of different molecular weights (MW) (such as 24,000, 40,000, 360,000, and 700,000 g. mole-1) were used for the interaction between CFH and PVP. The gradual increase in K b value and the complexation reaction was found to be much enhanced with the augmentation of the MW of PVP. The values of K b were also found to be increased with increasing temperatures as well as with the increase of electrolyte/acetic acid concentration. The Gibbs free energy of binding (∆G 0) values of the interaction process was negative which indicates the complex formation is thermodynamically spontaneous. The positive values of enthalpy (∆H 0) and entropy (∆S 0) of binding connote that the binding force for CFH-PVP complexation is hydrophobic in nature and the complexation is entropy controlled. The negative intrinsic enthalpy (∆H *,0) values indicate the high stability of CFH-PVP complexes. Molecular docking calculation discloses the existence of similar binding forces between CFH and PVP obtained by the analysis of experimental data from UV-visible spectroscopic method. The binding constant between CFH and BSA (K b ), quenching constant (K sv ), the number of binding sites (n), and the quenching rate constant (K q ) for the CFH-BSA system were also calculated. The values of K sv , K q , and n for the CFH-BSA system are lower in 0.05 mol L-1 urea solution and higher in PVP solutions compared to those of aqueous medium.

Nitrate is an abundant nutrient and electron acceptor throughout Earth’s biosphere. Virtually all nitrate in nature is produced by the oxidation of nitrite by the nitrite oxidoreductase (NXR) multiprotein complex. NXR is a crucial enzyme in the global biological nitrogen cycle, and is found in nitrite-oxidizing bacteria (including comammox organisms), which generate the bulk of the nitrate in the environment, and in anaerobic ammonium-oxidizing (anammox) bacteria which produce half of the dinitrogen gas in our atmosphere. However, despite its central role in biology and decades of intense study, no structural information on NXR is available. Here, we present a structural and biochemical analysis of the NXR from the anammox bacterium Kuenenia stuttgartiensis, integrating X-ray crystallography, cryo-electron tomography, helical reconstruction cryo-electron microscopy, interaction and reconstitution studies and enzyme kinetics. We find that NXR catalyses both nitrite oxidation and nitrate reduction, and show that in the cell, NXR is arranged in tubules several hundred nanometres long. We reveal the tubule architecture and show that tubule formation is induced by a previously unidentified, haem-containing subunit, NXR-T. The results also reveal unexpected features in the active site of the enzyme, an unusual cofactor coordination in the protein’s electron transport chain, and elucidate the electron transfer pathways within the complex., The oxidoreductase (NXR) multiprotein complex is a key enzyme in the nitrogen cycle. A detailed structural and biochemical characterization of NXR from the anammox bacterium Kuenenia stuttgartiensis shows that this complex is a filament-forming protein that catalysers both nitrite oxidation and nitrate reduction, and elucidates the mechanisms governing complex assembly and function.

Several bruchid species attack stored pulses including those belonging to Vigna species.Wild Vigna accessions offer a host of useful traits for improvement of their cultivated types and hence 42 endemic wild Vigna accessions from 13 species were screened against three cosmopolitan bruchid species viz., Callosobruchus maculatus (F.), C. chinensis L. and C. analis (F.). The accessions exhibited varied reactions to three bruchid species and were categorized based on two criteria-seed damage (SD) and susceptibility index (SI). Based on SD, IC251442 accession of V. umbellata was found highly resistant, whereas accessions of V. umbellata (IC251439, PRR 2007-2 and IC251440), V. vexillata (IC248326 and IC248343), V. trilobata (JAP10-5) and V. trineriva var. bournei (IC247407) were found resistant to C. maculatus. Likewise, the accessions of V. mungo var. mungo (IC251390, IC251385 and IC251387), V. vexillata (IC248326) and V. khandalensis (Kumur local) were found highly resistant, whereas the accessions V. umbellata (IC251440), V. mungo var. mungo (IC251397), V. vexillata (IC248343) and V. dalzelliana (IC247408) were found resistant to C. chinensis. Interestingly, none of the accessions was found either highly resistant/resistant to C. analis. Based on SI, several accessions (including those susceptible on the basis of SD) recorded lower SI (

UV–visible and florescence spectroscopic methods have been applied to analyze the binding capabilities of Ag+ ions and AgNPs with bovine serum albumin (BSA). UV–visible studies revealed that BSA acted as a ligand towards both Ag+ and AgNPs, and provides potential site for coordination. BSA fluorescence was quenched significantly by the quenchers (Ag+ and AgNPs). The binding constant, quenching constant, quantum yield, and coordination sites were evaluated. The Ag+ ions has strong binding and quenching efficiency towards BSA than that of AgNPs. The distance between the tryptophan and acceptor decreases from 127 A to 72 A with Ag+ ions from 0.6 to 20.0 × 10-5 mol/L. The energy transfer from BSA to ligand (Ag+ ions and AgNPs) follows a surface energy transfer process with a 1/d4 distance dependence. This is found to be according to strong coordinating affinity of metal ions with structural unit of protein, which helps to understand the mechanism of bacterial cell death in presence of metal ions and/or metal NPs.

To study the interaction of the series of pyridinium based gemini surfactants (GS) (referred to as m-Py-m, m = 14, 16); 4,4′-(propane-1,3-diyl)bis(1-(2-(tetradecyloxy)-2-oxoethyl) dipyridinium chloride (14-Py-14), and 4,4′-(propane-1,3-diyl) bis(1-(2-(hexadecyloxy)-2-oxoethyl)dipyridinium chloride (16-Py-16) with bovine serum albumin (BSA), various physicochemical and spectroscopic tools such as tensiometry, steady-state fluorescence, synchronous fluorescence, pyrene fluorescence, UV–visible, far-UV circular dichroism (CD) were utilized at physiological pH (7.4) and 298 K in combination with computational molecular modeling analysis. The tensiometric results show significant modifications in interfacial and thermodynamic parameters for m-Py-m GS upon BSA combination, deciphering the gemini surfactant-BSA interaction. Steady-state fluorescence analysis evaluates the structural alterations of BSA with the addition of m-Py-m GS. The plots of Stern-Volmer, modified Stern-Volmer, and thermodynamic parameters were used to determine the binding type of m-Py-m GS to BSA. The synchronous fluorescence spectra state a mild effect of gemini surfactants on the emission intensity of tyrosine (Tyr) residues, on the other hand, tryptophan (Trp) residues showed a significant effect. Post addition of GS, the plot of pyrene fluorescence reveals the mild micropolarity fluctuations via the probe (pyrene) molecules encapsulated in BSA. UV–visible experiments support the complex formation between the BSA and m-Py-m GS. Far-UV CD measurements revealed the modifications in the secondary structure of protein produced by m-Py-m GS. Furthermore, we also used the computational molecular modeling for attaining deep insight into BSA and m-Py-m GS binding and the results are supported with our experimental results.

Glutaredoxins (Grxs) are small (10–15 kDa) glutathione (GSH) - dependent redox proteins. The role of Grxs are well documented in tolerance to heavy metal stress in prokaryotic and mammalian systems and a few plant genera, but is poorly understood in plants against drought. In the present study, two rice glutaredoxin (Osgrx) genes (LOC_Os02g40500 and LOC_Os01g27140) responsible for tolerance against heavy metal stress have been studied for investigating their role against drought. Each glutaredoxin gene was over-expressed in Arabidopsis thaliana to reveal their role in drought stress. The relative expression of both Osgrx genes was higher in the transgenic lines. Transgenic lines of both Osgrxs showed longer roots, higher seed germination, and survival efficiency during drought stress. The physiological parameters (PN, gs, E, WUE, qP, NPQ and ETR), antioxidant enzymes (GRX, GR, GPX, GST, APX, POD, SOD, CAT, DHAR, and MDHAR), antioxidant molecules (ascorbate and GSH) and stress-responsive amino acids (cysteine and proline) levels were additionally increased in transgenic lines of both Osgrxs to provide drought tolerance. The outcomes from this study strongly determined that each Osgrx gene participated in the moderation of drought and might be utilized in biological engineering strategies to overcome drought conditions in different crops.

Glutaredoxins (Grxs) are small multifunctional redox proteins. Grxs have glutathione-dependent oxidoreductase activity in the presence of glutathione reductase and NADPH. The role of Grxs is well studied in heavy metal tolerance in prokaryotic and mammalian systems but not in plant genera. In the present study, a chickpea glutaredoxin (CaGrx) gene (LOC101493651) has been investigated against metal stress based on its primary screening in chickpea which revealed higher up-regulation of CaGrx gene under various heavy metals (AsIII-25 μM, AsV-250 μM, Cr(VI)-300 μM, and Cd-500 μM) stress. This CaGrx gene was overexpressed in Arabidopsis thaliana and investigated various biochemical and physiological performances under each metal stress. Transgenic plants showed significant up-regulation of the CaGrx gene during qRT-PCR analysis as well as longer roots, higher seed germination, and survival efficiency during each metal stress. The levels of stress markers, TBARS, H2O2, and electrolyte leakage were found to be less in transgenic lines as compared to WT revealed less toxicity in transgenics. The total accumulation of AsIII, AsV, and Cr(VI) were significantly reduced in all transgenic lines except Cd, which was slightly reduced. The physiological parameters such as net photosynthetic rate (PN), stomatal conductance (gs), transpiration (E), water use efficiency (WUE), photochemical quenching (qP), and electron transport rate (ETR), were maintained in transgenic lines during metal stress. Various antioxidant enzymes such as glutaredoxin (GRX), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione-S-transferase (GST), ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), antioxidant molecules (ascorbate, GSH) and stress-responsive amino acids (proline and cysteine) levels were significantly increased in transgenic lines which provide metal tolerance. The outcome of this study strongly indicates that the CaGrx gene participates in the moderation of metal stress in Arabidopsis, which can be utilized in biotechnological interventions to overcome heavy metal stress conditions in different crops.

Drought is one of the major abiotic stresses which negatively affect plant growth and crop yield. Metallothionein (MTs) is a low molecular weight protein, mainly involved in metal homeostasis, while, its role in drought stress is still to be largely explored. The present study was aimed to investigate the role of MT gene against drought stress. The chickpea MT based on its up-regulation under drought stress was overexpressed in Arabidopsis thaliana to explore its role in mitigation of drought stress. The total transcript of MT gene was up to 30 fold higher in transgenic lines. Arabidopsis plants transformed with MT gene showed longer roots, better efficiency of survival and germination, larger siliques and higher biomass compared to WT. The physiological variables (A, WUE, G, E, qP and ETR) of WT plants were reduced during drought stress which recovered in transgenic Arabidopsis lines. The enzymatic and non-enzymatic antioxidant (APX, GPX, POD, GR, GRX, GST, CAT, MDHAR, ASc and GSH) levels were also enhanced in transgenic lines to provide tolerance. Simultaneously, drought responsive amino acids, i.e. proline and cysteine contents were higher in transgenic lines. Overall, the results suggest that MT gene is actively involved in the mitigation of drought stress and could be the choice for genetic engineering strategy to overcome drought stress.

Pelodera scrofulata sp. nov. is characterized by cuticle with longitudinal lines and punctations; offset lip region with separate lips, wider in females, moderately developed cheilostom, presence of glandular bodies associated with uterus, tail conoid to spicate; males with spicules fused up to 60–68% from distal end; peloderan bursa with two precloacal papillae 9–12 μm apart and seven postcloacal pairs in the configuration of 1 + 1/4 + P + 2 + 1. Pelodera aligarhensis sp. nov. is characterized by sexual dimorphism in anterior body region; cuticle often with faint longitudinal lines; slightly offset lip region; tail cupola-shaped; males with spicules fused up to 20-25% from distal end; peloderan bursa; one precloacal pair at anterior level of cloaca; two precloacals shifted posteriad to the cloacal level; seven postcloacal pairs of genital papillae in 1/2 + (2 + P) + (3 + 1) configuration. Some new information is added to the description of Pelodera teres (Schneider, 1866) with special reference to labial...

Interaction of surfactants with biomacromolecules is an essential subject of biophysical chemistry to address their diverse applications in industry, biomedical, and cosmetic domains. In this context, we have examined the binding interactions of three ester-functionalized surfactants (m-E2-m) with bovine liver catalase (BLC, 10 μM) by employing a multi-technique approach. The m-E2-m geminis quench fluorescence intensity of BLC through static procedure. The binding ability of concerned gemini surfactants was found to be in the order 12-E2-12 (K b = 2.3 y 10 2 ) > 16-E2-16 (K b = 1.1 y 10 2 ) > 14-E2-14 (K b = 1.0 y 10 2 ). Quenching efficacy, as determined by K sv values, were observed as 12-E2-12 (3.0 y 10 2 ) > 16-E2-16 (1.4 y 10 2 ) > 14-E2-14 (1.0 y 10 2 ). The negative οG° b values (12-E2-12 (⿿13.48 kJ/mol) > 16-E2-16 (⿿11.65 kJ/mol) > 14-E2-14 (⿿11.41 kJ/mol)) indicate spontaneous nature of m-E2-m⿿BLC interactions. UV⿿vis spectroscopy, circular dichroism (CD) and micropolarity (F 1 /F 3 ) assessments indicate conformational changes in BLC upon m-E2-m combination. ITC confirms the stability of BLC upon gemini combination. Docking provides support to fluorescence results by presenting the localization site of m-E2-m surfactants near to aromatic residues (mainly Tyr, Trp and Phe). Moreover, since surfactant-protein interactions have essential miscellaneous implications, therefore, this study can be significant for industrial and biomedical realms.

Mungbean yellow mosaic India virus (MYMIV) is recognized as one of the most economically important viruses that affects several leguminous crops like mungbean, urdbean, cowpea, pigeonpea etc. and occurs in Indian subcontinent. In the present study, coat protein of MYMIV is being used to fi nd out highly suitable MHC binding peptides and epitopes. Thirty nine peptide regions were found to have high af fi nity to TAP binding peptides using cascade support vector machine (SVM). Few of these coat protein TAP transporters are 201- NRFFKVNNY with score 9.208, 108- KRFCIKSVY with score 8.817, 44-RWTNRPMWR with score 8.790, 134- NTVMFKLCR with score 8.672 and 41- KRRRWTNRP with score 8.498 where the scores are based on the average af fi nity of an amino acid at particular position. The SVM based method for prediction of promiscuous MHC Class II binders reported MHCII-IAb peptide regions, 30- PASAGGVPT, 127-IKSKNHTNT, 44-RWTNRPMWR, 6-YDTAFSTPI, (optimal score 1.220); MHCII-IAd peptide regions, 226-NALLLYMAC, 30-PASAGGVPT, 32-SAGGVPTNM, 236- HASNPVYAT, (optimal score 0.620); MHCII-IAg7 peptide regions, 13-PISNARRRL, 212- YNHQEAAKY, 221-ENHTENALL, 209-YVVYNHQEA, (optimal score 1.569) and MHCII- RT1.B peptide regions, 223- HTENALLLY, 188- TGGQYACKE, 168- TVKNDLRDR, 4-RTYDTAFST, (optimal score 0.932) as possible predicted binders from coat protein. The most suitable predicted segments in coat protein of MYMI virus for developing speci fi c antibodies found in this study are 56-FYRLYRSPDVPRGCEGPCKVQSF–78, 206-VNNYVVYNHQ-215 and 108-KRFCIKSVYITG-119. Fragments identi fi ed through this approach tend to be high- ef fi ciency binders, in which larger percentage of their atoms are directly involved in binding as compared to larger molecules. These fragments may, therefore, be used in cross protection and to develop begomovirus speci fi c antibodies that can be exploited in sero-diagnostics.

This study was made to find the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae in neonatal intensive care unit (NICU) and to identify the risk factors associated with the acquisition of these organisms. Risk factors associated with ESBL-producing E. coli and/or K. pneumoniae acquisition status of neonates were assessed. Of 253 neonates admitted, 238 entered the active surveillance system. ESBL-producing K. pneumoniae was responsible for 7 infections and 51 colonizations while ESBL-producing E. coli was responsible for 9 infections and 88 colonizations. Concurrent isolation of both the organisms occurred in 30 neonates. The logistic regression model identified 'length of stay in the NICU' as the single independent risk factor. Imipenem, cefepime and amikacin can be suggested as the drugs of choice in our study.

Antimicrobial activities of the crude ethanolic extracts of five plants were screened against multidrug resistant (MDR) strains of Escherichia coli, Klebsiella pneumoniae and Candida albicans. ATCC strains of Streptococcus mutans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus bovis, Pseudimonas aeruginosa, Salmonella typhimurium, Escherichia coli, Klebsiella pneumoniae and Candida albicans were also tested. The strains that showed resistance against the maximum number of antibiotics tested were selected for an antibacterial assay. The MDR strains were sensitive to the antimicrobial activity of Acacia nilotica, Syzygium aromaticum and Cinnamum zeylanicum, whereas they exhibited strong resistance to the extracts of Terminalia arjuna and Eucalyptus globulus. Community-acquired infections showed higher sensitivity than the nosocomial infections against these extracts. The most potent antimicrobial plant was A. nilotica (MIC range 9.75-313 microg/ml), whereas other crude plant extracts studied in this report were found to exhibit higher MIC values than A. nilotica against community acquired as well as nosocomial infection. This study concludes that A. nilotica, C. zeylanicum and S. aromaticum can be used against multidrug resistant microbes causing nosocomial and community acquired infections.

Summary Background The β-lactam antibiotics, in combination with aminoglycosides, are among the most widely prescribed antibiotics. However, because of extensive and unnecessary use, resistance to these drugs continues to increase. In recent years, resistance in the Indian bacterial population has increased markedly, the majority showing complex mechanisms. Due to increased transcontinental movement of the human population, it would be wise to know the prevalence and resistance complexity of these strains, well in advance, in order to formulate a policy for empirical therapy. Methods One hundred and eighty-one isolates of Escherichia coli and 61 isolates of Klebsiella pneumoniae obtained from 2655 non-repeat samples of pus (912) and urine (1743) were studied, and their resistance rates and patterns were noted. The isolates were analyzed for prevalent aminoglycoside and cephalosporin resistance phenotypes and for the presence of extended spectrum β-lactamase (ESBL) and AmpC enzymes by spot-inoculation and modified three-dimensional tests developed in our laboratory. Fourteen isolates of E. coli and six of K. pneumoniae , resistant to all of the antibiotics tested, were selected for plasmid screening, curing, and transconjugation experiments, and for comparative evaluation of the double disk synergy test (DDST) and modified three-dimensional test (TDT) for detection of β-lactamases. Results Urinary E. coli isolates showed maximum susceptibility to amikacin (57.1%), followed by tobramycin (38.5%) and gentamicin (31.9%). Eighteen (19.8%) isolates were susceptible to cefotaxime, whereas 11 (12.1%) were susceptible to ceftriaxone. The K. pneumoniae isolates from urine samples showed maximum susceptibility to tobramycin (63.6%) followed by amikacin (54.5%). Of the K. pneumoniae isolates, 31.8% were susceptible to cefotaxime and 13.6% were susceptible to ceftriaxone. A more or less similar trend of antibiotic susceptibility was noted in E. coli and K. pneumoniae isolates from pus samples. Twenty-six (14.4%) E. coli and 15 (24.6%) K. pneumoniae isolates were found to be ESBL-producers by NCCLS-ESBL phenotypic confirmatory test. Eighteen (9.9%) E. coli and 19 (31.1%) K. pneumoniae isolates were found to be AmpC enzyme-producers by our modified TDT. The simultaneous occurrence of ESBL and AmpC enzymes was noted in 7.7% and 9.8% isolates of E. coli and K. pneumoniae , respectively. Conclusions The prevalence of multidrug-resistant bacterial isolates is quite high in our bacterial population. On comparative evaluation of DDST and TDT in resistant isolates, TDT was found to be the better method, detecting ESBLs in 80% of isolates compared to 15% with DDST. A 19.9-kb plasmid was consistently present in all the screened isolates of E. coli and K. pneumoniae , and was inferred to encode cefoxitin and tetracycline resistance based on curing and transconjugation experiments.

Summary The nucleotide and amino acid sequences of the movement protein (NSm) genes of five isolates of Groundnut bud necrosis virus (GBNV) originating from different hosts and parts of India such as cowpea and tomato from Kerala, groundnut from Tamil Nadu, and potato from Madhya Pradesh and Rajasthan were determined and compared to the known NSm sequences. Sequence analysis revealed that the NSm genes of GBNV isolates were identical in length (924 bp encoding 307 amino acids). GBNV isolates shared maximum identity (98–100%) at amino acid levels with GBNV-Type isolate, while 82–83% and 34–65% amino acid sequence identities were observed with Watermelon silver mottle virus and other Tospoviruses respectively. The NSm genes among GBNV isolates originating from different hosts and locations appeared highly conserved (93–100%), suggesting their common origin.

Integrons have been observed to be frequently associated with uropathogenic bacteria. This study aimed at 1) determining the prevalence of class 1 integrons among ESBLl-producing uropathogenic Escherichia coli, and 2) analyzing resistance genes associated with different phylogenetic groups of the integron-positive isolates with special reference to bla(CTX-M) and bla(TEM). Twenty-three ESBL-producing E. coli were studied. Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) displayed 14 major patterns. Pulse field Gel electrophoresis-typing of 8 randomly selected integron-positive strains ruled out any correlation between genotype and antibiotype. Genomic DNA from 14 strains was PCR-positive for class 1 integrons, bla(CTX-M-15) and bla(TEM-1)-like genes. Integron-sequencing revealed "aadA5-dfrA17-dfrA7" as the most prevalent gene cassette. Our findings unveil the increasing role of the bla(CTX-M) genes in antibiotic resistance and emphasize on the significance of appropriate empirical treatment for Urinary tract infections. Moreover, this is the first study which reports bla(CTX-M) located on genomic DNA of bacteria from India.

Groundnut bud necrosis virus (GBNV) is recognized as one of the most economically important viruses and is known to affect several crops including peanut, potato, tomato and soybean. For managing plant virus diseases, determination of their causal agents' identity at an early stage of crop is a pre-requisite. In the present study, NSm protein of GBNV has been used to predict out MHC binding peptides and epitopes that are highly suitable for antigenicity. Eighteen peptide regions were found to have high affinity. Few of these NSm protein TAP transporters are 126- RRYMHISRL with score 11.638, 125- NRRYMHISR with score 10.280, 46- AIMNKAKTL with score 7.762, 120- PTWNSNRRY with score 7.632 and 171- ASLKDPMCF with score 7.277. The support vector machine (SVM) based approach predicted MHCII-IAb peptide regions, 45- SAIMNKAKT, 151- ASLIDPNKM, 23- PAVKKENNR, 229- PIAAENNTC, (optimal score 0.938); MHCII-IAd peptide regions, 208- YAKGVGFAS, 101- NDSLVGNGN, 55- NGKQYVSSG, 63-GDSSVLGTY, (optimal score 0.852); MHCII-IAg7 peptide regions, 277- LQKAAERLA, 145- SKNNVKASL, 228- TPIAAENNT, 276- SLQKAAERL, (optimal score 1.640); and MHCII- RT1.B peptide regions, 193- TPKQCMQLN, 195- KQCMQLNLT, 246- KVIQSAALI, 166- IISRQASLK, (optimal score 0.800) as binders from NSm protein. The most suitable predicted segments in NSm protein of GBNV virus found in the study are 164- KIIISRQASLKDPMCFIFHLNWS- 186 and 237-CDVVPINRAKVIQSAALIEACKLMIP-262. These two fragments, obtained from non-structural movement protein with average propensity 1.016, are high-efficiency binders and may, therefore be used in cross protection to provide resistance against GBNV and develop GBNV specific antibodies that can be exploited in sero-diagnostics.

Background: Antibiotic resistance is common among bacterial pathogens associated with both community acquired and nosocomial infections. In view of the present problem of drug resistance we investigated the prevalence of methicillin resistant Staphylococcus aureus (MRSA) and amplified the mecA gene in the isolates from the hand swabs of the hospital personnel. Methodology: The nuc gene was amplified to characterize these isolates at species level. The S. aureus isolates were analyzed for their susceptibility to different classes of antibiotics using the disk diffusion method. The spot inoculation test was performed to detect methicillinase production in these isolates. Results: In the screened isolates of S. aureus, 14.2 and 15 kb of plasmids were present. These isolates showed pronounced resistance against β-lactam antibiotics including second- and third-generation cephalosporins, aminoglycosides, macrolides and fluoroquinolone. Some of the isolates included in this study were resistant to three or more antibiotics. Expression of methicillinase was detected by spot inoculation test, and a few of the isolates were found to produce methicillinase. Moreover, mecA gene was also amplified. Of 17 isolates only 7 showed presence of mecA gene. Conclusion: This study highlights the emerging trend of multiple drug resistance in S. aureus strains isolated from hospital personnel working in a premier hospital in North India.

A systematic investigation of the effect of polyethylene glycols, salts, and alcohols on the trichloroacetic acid (TCA)-induced state of ferricytochrome c was made using various spectroscopic techniques. Native cytochrome c (Cyt c) has a fluorescence maximum at 335 nm, where as the TCA-induced state of Cyt c has a red shift of 7 nm with enhanced fluorescence intensity. The near-and far-UVCD spectra showed a significant loss of tertiary and secondary structure, although the protein is relatively less unfolded as compared with a conformation at pH 2.0. Addition of 70% (v/v) polyols to TCA (3.3 mM)-induced state of Cyt c resulted in increased 1-anilino-8-naphthalene sulfonate binding and increased mean residue ellipticity at 222 nm, indicating increase in compactness with enhanced exposure of hydrophobic surface area. Also, the stabilizing effect of salts and alcohols on the TCA-induced state was studied and compared with their effect on trifluoroacetic acid-unfolded state of Cyt c. Among all the polyols, salts, and alcohols studied, PEG-400, K3[Fe(CN)6], and butanol were the most efficient in inducing secondary structure in TCA-induced state as examined by the above-mentioned spectroscopic techniques. For salts, the efficiency in inducing the secondary structure followed the order K3[Fe(CN)6] > KClO4 > K2SO4 > KCl. For alcohols, this order was found to be as follows: butanol > propanol > ethanol > methanol.

A number of wild relatives of the two cultivated species of Vigna ( V. mungo and V. radiata ) are known to occur in different parts of India and are exploited for their useful genes, including resistance to yellow mosaic disease (YMD). There…

Pisum sativum is an important cool season legume crop cultivated under irrigated conditions in many parts of India particularly in the northern plains. It is used as a food grain or as vegetable. In India, there are few reports of viruses…