scholarly journals Engineering Rhizobial Bioinoculants: A Strategy to Improve Iron Nutrition

2013 ◽  
Vol 2013 ◽  
pp. 1-15 ◽  
Author(s):  
S. J. Geetha ◽  
Sanket J. Joshi
Keyword(s):  
Genetic Engineering ◽  
Plant Growth ◽  
Outer Membrane ◽  
Iron Transport ◽  
Iron Acquisition ◽  
Membrane Receptors ◽  
Iron Nutrition ◽  
Nodule Occupancy ◽  
Rhizobial Strain ◽  
Hydroxamate Siderophores

Under field conditions, inoculated rhizobial strains are at a survival disadvantage as compared to indigenous strains. In order to out-compete native rhizobia it is not only important to develop strong nodulation efficiency but also increase their competence in the soil and rhizosphere. Competitive survival of the inoculated strain may be improved by employing strain selection and by genetic engineering of superior nitrogen fixing strains. Iron sufficiency is an important factor determining the survival and nodulation by rhizobia in soil. Siderophores, a class of ferric specific ligands that are involved in receptor specific iron transport into bacteria, constitute an important part of iron acquisition systems in rhizobia and have been shown to play a role in symbiosis as well as in saprophytic survival. Soils predominantly have iron bound to hydroxamate siderophores, a pool that is largely unavailable to catecholate-utilizing rhizobia. Outer membrane receptors for uptake of ferric hydroxamates include FhuA and FegA which are specific for ferrichrome siderophore. Increase in nodule occupancy and enhanced plant growth of thefegAandfhuAexpressing engineered bioinoculants rhizobial strain have been reported. Engineering rhizobia for developing effective bioinoculants with improved ability to utilize heterologous siderophores could provide them with better iron acquisition ability and consequently, rhizospheric stability.

scholarly journals High-Throughput Screening Assay for Inhibitors of TonB-Dependent Iron Transport

2015 ◽  
Vol 21 (3) ◽  
pp. 316-322 ◽  
Author(s):  
Mathew Hanson ◽  
Lorne D. Jordan ◽  
Yan Shipelskiy ◽  
Salete M. Newton ◽  
Phillip E. Klebba
Keyword(s):  
Outer Membrane ◽  
High Throughput ◽  
High Throughput Screening ◽  
Iron Transport ◽  
Iron Acquisition ◽  
Metabolic Inhibitors ◽  
Screening Assay ◽  
Gram Negative ◽  
Microtiter Plates ◽  
High Throughput Screening Assay

The TonB-dependent Gram-negative bacterial outer membrane protein FepA actively transports the siderophore ferric enterobactin (FeEnt) into the periplasm. We developed a high-throughput screening (HTS) assay that observes FeEnt uptake through FepA in living Escherichia coli, by monitoring fluorescence quenching that occurs upon binding of FeEnt, and then unquenching as the bacteria deplete it from solution by transport. We optimized the labeling and spectroscopic methods to screen for inhibitors of TonB-dependent iron uptake through the outer membrane. The assay works like a molecular switch that is on in the presence of TonB activity and off in its absence. It functions in 96-well microtiter plates, in a variety of conditions, with Z factors of 0.8–1.0. TonB-dependent iron transport is energy dependent, and the inhibitory effects of the metabolic inhibitors carbonyl cyanide m-chlorophenylhydrazone, 2,4-dinitrophenol, azide, cyanide, and arsenate on FeEnt uptake were readily detected by the assay. Because iron acquisition is a determinant of bacterial pathogenesis, HTS with this method may identify inhibitors that block TonB function and constitute novel therapeutics against infectious disease caused by Gram-negative bacteria.

scholarly journals Study on RNA Regulating Iron Transport in Outer Membrane Vesicles of Hypervirulent Klebsiella Pneumoniae

2020 ◽  
Author(s):  
Mao Zhou ◽  
Siyi Wang ◽  
You Lan ◽  
Xin Li ◽  
Xuan Liu ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Outer Membrane ◽  
Iron Transport ◽  
High Throughput Sequencing ◽  
Iron Acquisition ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Transporter Family ◽  
Iron Deficient ◽  
Deficient Medium

Abstract Background: The iron acquisition ability of hypervirulent Klebsiella pneumoniae (hvKP) is an important part of its super virulence mechanism, increasing studies have proved that outer membrane vesicles (OMVs) are involved in the iron acquisition process of bacteria. Thus, we compared the difference in RNA expression in OMVs of hvKP in iron-rich and iron-deficient medium, and explore the possible mechanism of RNA in OMVs involved in hvKP iron acquisition. Results: The results of high-throughput sequencing showed that in iron-deficient medium, there were 239 up-regulated and 89 down-regulated mRNAs in OMVs of hvKP, of which 20 mRNAs related to iron transport was up-regulated, mainly including siderophore synthesis and receptor genes, ATP binding cassette transporter family and iron sulfur cluster. Only two of the differential ncRNAs that regulate these mRNAs are up-regulated, which are lncRNAs.Conclusion: We demonstrated that mRNA and lncRNA in OMVs were directly or indirectly involved in the iron acquisition mechanism of hvKP under iron deficiency environment, which enhanced the adaptive survival ability of hvKP. It provided a basis for further exploring the iron acquisition mechanism of OMVs involved in hvKP.

scholarly journals Yersinia pestis TonB: Role in Iron, Heme, and Hemoprotein Utilization

2003 ◽  
Vol 71 (7) ◽  
pp. 4159-4162 ◽  
Author(s):  
Robert D. Perry ◽  
Jessica Shah ◽  
Scott W. Bearden ◽  
Jan M. Thompson ◽  
Jacqueline D. Fetherston
Keyword(s):  
Yersinia Pestis ◽  
Outer Membrane ◽  
Transport System ◽  
Iron Transport ◽  
Membrane Receptors ◽  
Heme Transport ◽  
Iron Heme ◽  
Manganese Transport ◽  
Iron Utilization ◽  
Iron And Manganese

ABSTRACT In Yersinia pestis, the siderophore-dependent yersiniabactin (Ybt) iron transport system and heme transport system (Hmu) have putative TonB-dependent outer membrane receptors. Here we demonstrate that hemin uptake and iron utilization from Ybt are TonB dependent. However, the Yfe iron and manganese transport system does not require TonB.

scholarly journals Genes Essential to Iron Transport in the Cyanobacterium Synechocystis sp. Strain PCC 6803

2001 ◽  
Vol 183 (9) ◽  
pp. 2779-2784 ◽  
Author(s):  
Hirokazu Katoh ◽  
Natsu Hagino ◽  
Arthur R. Grossman ◽  
Teruo Ogawa
Keyword(s):  
Iron Uptake ◽  
Iron Transport ◽  
Ferric Iron ◽  
Iron Acquisition ◽  
Complete Medium ◽  
Iron Starvation ◽  
Transporter Family ◽  
Genes Encoding ◽  
In Cells ◽  
Pcc 6803

ABSTRACT Genes encoding polypeptides of an ATP binding cassette (ABC)-type ferric iron transporter that plays a major role in iron acquisition inSynechocystis sp. strain PCC 6803 were identified. These genes are slr1295, slr0513, slr0327, and recently reportedsll1878 (Katoh et al., J. Bacteriol. 182:6523–6524, 2000) and were designated futA1, futA2, futB, andfutC, respectively, for their involvement in ferric iron uptake. Inactivation of these genes individually or futA1and futA2 together greatly reduced the activity of ferric iron uptake in cells grown in complete medium or iron-deprived medium. All the fut genes are expressed in cells grown in complete medium, and expression was enhanced by iron starvation. ThefutA1 and futA2 genes appear to encode periplasmic proteins that play a redundant role in iron binding. The deduced products of futB and futC genes contain nucleotide-binding motifs and belong to the ABC transporter family of inner-membrane-bound and membrane-associated proteins, respectively. These results and sequence similarities among the four genes suggest that the Fut system is related to the Sfu/Fbp family of iron transporters. Inactivation of slr1392, a homologue offeoB in Escherichia coli, greatly reduced the activity of ferrous iron transport. This system is induced by intracellular low iron concentrations that are achieved in cells exposed to iron-free medium or in the fut-less mutants grown in complete medium.

scholarly journals Scope and future perspectives of phytoremediation

2021 ◽  
Vol 16 (AAEBSSD) ◽  
pp. 77-85
Author(s):  
Sridevi Tallapragada ◽  
Rajesh Lather ◽  
Vandana ◽  
Gurnam Singh
Keyword(s):  
Heavy Metals ◽  
Heavy Metal ◽  
Genetic Engineering ◽  
Plant Growth ◽  
Mycorrhizal Fungi ◽  
Large Scale ◽  
Negative Impact ◽  
Heavy Metal Accumulation ◽  
Plant Growth Promoting Bacteria ◽  
Runoff Water

Phytoremediation is the plant-based technology that has emerged as a novel cost effective and ecofriendly technology in which green plants are used for extraction, sequestration and/or detoxification of the pollutants. Plants possess the natural ability to degrade heavy metals and this property of plants to detoxify contaminants can be used by genetic engineering approach. Currently, the quality of soil and water has degraded considerably due heavy metal accumulation through discharge of industrial, agricultural and domestic waste. Heavy metal pollution is a global concern and a major health threat worldwide. They are toxic, and can damage living organisms even at low concentrations and tend to accumulate in the food chain. The most common heavy metal contaminants are: As, Cd, Cr, Cu, Hg, Pb and Zn. High levels of metals in soil can be phytotoxic, leading to poor plant growth and soil cover due to metal toxicity and can lead to metal mobilization in runoff water and thus have a negative impact on the whole ecosystem. Phytoremediation is a green strategy that uses hyperaccumulator plants and their rhizospheric micro-organisms to stabilize, transfer or degrade pollutants in soil, water and environment. Mechanisms used to remediate contaminated soil includes phytoextraction, phytostabilization, phytotransformation, phytostimulation, phytovolatilization and rhizofiltration. Traditional phytoremediation method presents some limitations regarding their applications at large scale, so the application of genetic engineering approaches such as transgenic transformation, nanoparticles addition and phytoremediation assisted with phytohormones, plant growth-promoting bacteria and Arbuscular mycorrhizal fungi (AMF) inoculation has been applied to ameliorate the efficacy of plants for heavy metals decontamination. In this review, some recent innovative technologies for improving phytoremediation and heavy metals toxicity and their depollution procedures are highlighted.

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