The rhizosphere offers a quintessential habitat for the microbial communities and facilitates a variety of plant-microbe interactions. Members of the genus Bacillus constitute an important group of ...plant growth promoting rhizobacteria (PGPR), which improve growth and yield of crops. In a total of 60 bacterial isolates from the tomato rhizosphere, 7 isolates were selected based on distinct morphological characteristics and designated as tomato rhizosphere (TRS) isolates with a number suffixed viz., TRS-1, 2, 3, 4, 5, 7, and TRS-8. All the seven isolates were Gram positive, with in vitro plant growth promoting (PGP) traits like phosphate and zinc solubilization, and also produced indoleacetic acid (IAA), phytase, siderophore, hydrogen cyanide (HCN), and 1-aminocyclopropane-1-carboxylate (ACC) deaminase, besides being antagonistic to other microbes and formed biofilm. The seven isolates belonged to the genus Bacillus as per the 16S rDNA sequence analysis. Phylogenetic tree grouped the isolates into four groups, while BOX-PCR fingerprinting allowed further differentiation of the seven isolates. The PGP activity of the isolates was measured on tomato seedlings in plant tissue culture and greenhouse assays. A significant increase in root colonization was observed over 15 days with all the isolates. Greenhouse experiments with these isolates indicated an overall increase in the growth of tomato plants, over 60 days. Isolates TRS-7 and TRS-8 were best plant growth promoters among the seven isolates, with a potential as inoculants to increase tomato productivity.
Microbiology; Agricultural Soil Science; Rhizosphere; Plant Growth; Bacteria; Microorganism; PGPR; tomato; Bacillus spp.; BOX-PCR
Neuroinflammation associated with Japanese encephalitis (JE) is mainly due to the activation of glial cells with subsequent release of proinflammatory mediators from them. The recognition of viral ...RNA, in part, by the pattern recognition receptor retinoic acid-inducible gene I (RIG-I) has been indicated to have a role in such processes. Even though neurons are also known to express this receptor, its role after JE virus (JEV) infections is yet to be elucidated.
Upon infecting murine neuroblastoma cells and primary cortical neurons with JEV the expression profile of key proinflammatory cyto/chemokines were analyzed by qRT-PCR and bead array, both before and after ablation of RIG-I. Immunoblotting was performed to evaluate the levels of key molecules downstream to RIG-I leading to production of proinflammatory mediators. Changes in the intracellular viral antigen expression were confirmed by intracellular staining and immunoblotting. JEV infection induced neuronal expression of IL-6, IL-12p70, MCP-1, IP-10 and TNF-α in a time-dependent manner, which showed significant reduction upon RIG-I ablation. Molecules downstream to RIG-I showed significant changes upon JEV-infection, that were modulated following RIG-I ablation. Ablation of RIG-I in neurons also increased their susceptibility to JEV.
In this study we propose that neurons are one of the potential sources of proinflammatory cyto/chemokines in JEV-infected brain that are produced via RIG-I dependent pathways. Ablation of RIG-I in neurons leads to increased viral load and reduced release of the cyto/chemokines.
Microorganisms represent a substantial portion of the earth's biodiversity and biomass, and the plant rhizosphere is an innate reservoir teeming with heterogeneous microbes predominated by bacterial ...communities. Rhizospheric microbial diversity (genetic, phenotypic, and metabolic) has been extensively studied to understand the key ecological roles played by the microbial members, including plant growth promotion. The application of 16S rRNA gene sequencing and next-generation sequencing (NGS) technologies has revolutionized the discovery of novel bacterial groups that have remained undetected by traditional cultivation-based approaches. Such technological advancements have opened new vistas in our current understanding of predominant but concealed and missed bacterial diversity referred to as difficult-to-culture bacterial lineages, especially the predominant phyla Acidobacteria, Verrucomicrobia, Planctomycetes, and Gemmatimonadetes. Regardless of their ubiquity and prevalence, little is known about their ecophysiology because of the non-availability of culturable members. More recently, there has been increased interest in understanding the cosmopolitan distribution and diversity of the difficult-to-culture bacteria, focusing on their role in driving complex plant-microbial interactions and mobilizing nutrients in soil and their potential as sources of novel bioactive metabolites. As an initial step, we review the distribution and significance of such bacterial phyla in soil, their ecophysiological roles, and their hidden plant growth promoting potential. The ability to select and deploy plant probiotic bacteria from the difficult-to-culture fraction of the bacterial community might open new avenues for improving crop health.
Japanese Encephalitis virus (JEV) is a common cause of acute and epidemic viral encephalitis. JEV infection is associated with microglial activation resulting in the production of pro-inflammatory ...cytokines including Interleukin-1 β (IL-1β) and Interleukin-18 (IL-18). The Pattern Recognition Receptors (PRRs) and the underlying mechanism by which microglia identify the viral particle leading to the production of these cytokines is unknown.
For our studies, we have used murine model of JEV infection as well as BV-2 mouse microglia cell line. In this study, we have identified a signalling pathway which leads to the activation of caspase-1 as the key enzyme responsible for the maturation of both IL-1β and IL-18 in NACHT, LRR and PYD domains-containing protein-3 (NLRP3) dependent manner. Depletion of NLRP3 results in the reduction of caspase-1 activity and subsequent production of these cytokines.
Our results identify a mechanism mediated by Reactive Oxygen Species (ROS) production and potassium efflux as the two danger signals that link JEV infection to caspase-1 activation resulting in subsequent IL-1β and IL-18 maturation.
Soils in the Lower Swansea Valley, (United Kingdom) contain elevated level of metals, enough to cause direct or indirect effects on human health. This study assesses the severity of soil pollution ...and bioavailability of Cu and other metals (Ni, Zn, Co, Pb and Cr) in soils with various distances from a Ni refinery. We compare Cu concentrations in operationally defined soil fractions (bioavailable, bound to Fe/Mn oxide and incorporated in organic matter) with other metals (Ni, Zn, Pb, Co, Cr) usually occurring in ores used in metallurgic processes and report their pollution and geoaccumulation indices (PI and
I
geo
). Further, we use Cu stable isotope ratios (
δ
65
Cu) to trace the fate and mobility of Cu in soils. Our data suggest a point source of contamination for some of the heavy metals including Ni (
I
geo
= 1.9), Zn (
I
geo
= 0.28) and Cu (
I
geo
= 3.6) near the Ni refinery. However, Co (
I
geo
= 0.15) and Pb (
I
geo
= 3.3) contaminations are likely to be linked to different sources. No elevated Cr levels (
I
geo
= -0.07) occur in any of the studied soils. All soil metals are predominantly associated with organic matter (>50%) which reduces their bioavailibility and thus their risk for ecological and human health. The Cu isotope data show that Cu in soil organic matter is enriched in
65
Cu, while the lighter isotopes (
63
Cu) remain in the dissolved bioavailable Cu fraction (Δ
65
Cu
organic-bioavailable
is +0.12 ± 0.13‰). This suggests the preferential complexation of
65
Cu with soil organic matter after dissolution of Cu deposited to the soil. Thus, Cu isotope data can effectively indicate pathways of metal migration in polluted soils.
•1-butyl-3-methylimidazolium bromide inhibits amyloid fibrillation in lysozyme.•Thioflavin T assay showed that fibrillation was suppressed.•Changes in the surface hydrophobicity of lysozyme were ...arrested in the presence of the ionic liquid.•AFM imaging showed that quantity of fibrils formed was reduced by the ionic liquid.
Many proteins can abnormally fold to form pathological amyloid deposits/aggregates that are responsible for various degenerative disorders called amyloidosis. Here we have examined the anti-amyloidogenic potency of an ionic liquid, 1-butyl-3-methylimidazolium bromide, using lysozyme as a model system. Thioflavin T fluorescence assay demonstrated that the ionic liquid suppressed the formation of lysozyme fibrils significantly. This observation was further confirmed by the Congo red assay. Fluorescence microscopy, intrinsic fluorescence studies, nile red fluorescence assay, ANS binding assay and circular dichroism studies also testified diminishing of the fibrillogenesis in the presence of ionic liquid. Formation of amyloid fibrils was also characterized by α to β conformational transition. From far-UV circular dichroism studies it was observed that the β-sheet content of the lysozyme samples decreased in the presence of the ionic liquid which in turn implied that fibrillogenesis was supressed by the ionic liquid. Atomic force microscopy imaging unequivocally established that the ionic liquid attenuated fibrillogenesis in lysozyme. These results may be useful for the development of more effective therapeutics for amyloidosis.
Cross-border data sharing for knowledge generation is a challenging research direction since an application may access personal data stored in countries different from the one where the application ...is accessed from. In this article, we propose a cross-border data sharing platform where a global cloud is built atop multiple security gateways that are set up in different countries. Once an application requests access to data from a particular country or region, the global cloud collects the data stored in local data hubs through that region's security gateway. While transferring the data to the global cloud, the security gateway records this transfer information on a blockchain maintained by the global cloud. When an application reports any misbehavior (e.g., providing wrong data type or incorrect data) against a security gateway, the global cloud verifies the claim by auditing the blockchain and punishes the misbehaving security gateway if the claim is true. In the case of false misbehavior report, the application itself will be punished by the global cloud. Thus, our platform provides an accountable data sharing function using blockchain that relies on a relaxed trust assumption on the data providers. We include five algorithms to handle data access request, data sharing, blockchain transaction, detecting, and punishing misbehaving entities. In the algorithms, we also introduce how the transaction takes place in the platform. Thus, the proposed platform is able to handle misbehaving data sender, data receiver, or any entity participating in the platform. We analyze our platform empirically by showing different graphs, which have been generated by a number of experiments on blockchain environment. We also delineate how the multilayer of signature (Elliptic Curve Digital Signature Algorithm) acts in our platform.
Host factors provide critical support for every aspect of the virus life cycle. We recently identified the valosin-containing protein (VCP)/p97, an abundant cellular ATPase with diverse cellular ...functions, as a host factor important for Japanese encephalitis virus (JEV) replication. In cultured cells, using siRNA-mediated protein depletion and pharmacological inhibitors, we show that VCP is crucial for replication of three flaviviruses: JEV, Dengue, and West Nile viruses. An FDA-approved VCP inhibitor, CB-5083, extended survival of mice in the animal model of JEV infection. While VCP depletion did not inhibit JEV attachment on cells, it delayed capsid degradation, potentially through the entrapment of the endocytosed virus in clathrin-coated vesicles (CCVs). Early during infection, VCP-depleted cells showed an increased colocalization of JEV capsid with clathrin, and also higher viral RNA levels in purified CCVs. We show that VCP interacts with the JEV nonstructural protein NS5 and is an essential component of the virus replication complex. The depletion of the major VCP cofactor UFD-1 also significantly inhibited JEV replication. Mechanistically, thus, VCP affected two crucial steps of the JEV life cycle - nucleocapsid release and RNA replication. Our study establishes VCP as a common host factor with a broad antiviral potential against flaviviruses.
JEV is the leading cause of viral encephalitis epidemics in South-east Asia, affecting majorly children with high morbidity and mortality. Identification of host factors is thus essential for the rational design of anti-virals that are urgently need as therapeutics. Here we have identified the VCP protein as one such host-factor. This protein is highly abundant in cells and engages in diverse functions and cellular pathways by its ability to interact with different co-factors. Using siRNA mediated protein knockdown, we show that this protein is essential for release of the viral RNA into the cell so that it can initiate replication. The protein plays a second crucial role for the formation of the JEV replication complex. FDA-approved drugs targeting VCP show enhanced mouse survival in JE model of disease, suggesting that this could be a druggable target for flavivirus infections.
•PRRs signaling are important in initiating innate antiviral and proinflammatory response following flavivirus-infection.•Flaviviruses have devised various strategies to evade PRR signaling.•Mainly ...TLR3 and TLR7 are involved in sensing of flaviviruses.•NLRs and CLRs have also been implicated in innate immune response against flaviviruses.
The flaviviral encephalitis has now become a major health concern in global scale. The efficient detection of viral infection and induction of the innate antiviral response by host's innate immune system are crucial to determine the outcome of infection. The intracellular pattern recognition receptors TLRs, RLRs, NLRs and CLRs play a central role in detection and initiation of robust antiviral response against flaviviral infection. Both cytoplasmic RLRs, RIG-I and MDA5 have been shown to be implicated in sensing flaviviral genomic RNA. Similarly among TLRs mainly TLR3 and TLR7 are known to respond in flaviviral infections as they are known to sense dsRNA and ssRNA moiety as their natural cognate ligand. Several studies have also shown the roles of NLRs and CLRs in mounting an innate antiviral response against flavivirus but, it is yet to be completely understood. Until now only few reports have implicated NLRs and CLRs in induction of antiviral and proinflammatory state following flaviviral infection. The current review therefore aims to comprehensively analyze past as well as current understanding on the role of PRRs in flaviviral infections.
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