scholarly journals Rapid Colorimetric Quantification of Lipo-chitooligosaccharides from Mesorhizobium loti and Sinorhizobium meliloti

2002 ◽  
Vol 15 (9) ◽  
pp. 859-865
Author(s):  
Joachim Goedhart ◽  
Jean-Jacques Bono ◽  
Theodorus W. J. Gadella
Keyword(s):  
Methylene Blue ◽  
Organic Phase ◽  
Sinorhizobium Meliloti ◽  
Sodium Dodecyl ◽  
Ion Pair ◽  
Nod Factors ◽  
Two Phase ◽  
Nod Factor ◽  
Mesorhizobium Loti ◽  
Hydrolysis Of

Nod factors are lipids with a chitinlike headgroup produced by gram-negative Rhizobium bacteria. These lipo-chitooligosaccharides (LCOs) are essential signaling molecules for accomplishing symbiosis between the bacteria and roots of legume plants. Despite their important role in the Rhizobium-legume interaction, no fast and sensitive Nod factor quantification methods exist. Here, we report two different quantification methods. The first is based on the enzymatic hydrolysis of Nod factors to release N-acetylglucosamine (GlcNAc), which can subsequently be quantified. It is shown that the degrading enzyme, glusulase, releases exactly two GlcNAc units per pentameric nodulation factor from Mesorhizobium loti factor, allowing quantification of LCOs from Mesorhizobium loti. The second method is based on a specific type of Nod factors that are sulfated on the reducing GlcNAc, allowing quantification analogous to the quantification of sulfolipids. Here, a two-phase extraction method is used in the presence of methylene blue, which specifically forms an ion pair with sulfated lipids. The blue ion pair partitions into the organic phase, after which the methylene blue signal can be quantified. To enable Nod factor quantification with this method, the organic phase was modified and the partitioning was evaluated using fluorescent and radiolabeled sulfated Nod factors. It is shown that sulfated LCOs can be quantified with this method, using sodium dodecyl sulfate for calibration. Both methods allow Nod factor quantification in parallel enabling a fast and easy detection of nanomole quantities of Nod factors. Accurate Nod factor quantification will be crucial for characterization and cross-comparison of the affinity for Nod factors of newly identified Nod factor binding proteins or putative Nod factor receptors.

scholarly journals Strain-Ecotype Specificity in Sinorhizobium meliloti-Medicago truncatula Symbiosis Is Correlated to Succinoglycan Oligosaccharide Structure

2007 ◽  
Vol 189 (21) ◽  
pp. 7733-7740 ◽  
Author(s):  
Senay Simsek ◽  
Tuula Ojanen-Reuhs ◽  
Samuel B. Stephens ◽  
Bradley L. Reuhs
Keyword(s):  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Donor Strain ◽  
Factor Activity ◽  
Nod Factors ◽  
Strain Specificity ◽  
Oligosaccharide Structure ◽  
Nod Factor ◽  
Structural Nature ◽  
Molecular Signals

ABSTRACT Molecular signals, including Nod factors and succinoglycan, are necessary for the establishment of nitrogen-fixing nodules (Fix+) in Medicago truncatula-Sinorhizobium meliloti symbiosis. This report shows that M. truncatula-S. meliloti interactions involve ecotype-strain specificity, as S. meliloti Rm41 and NRG247 are Fix+ (compatible) on M. truncatula A20 and Fix− (incompatible) on M. truncatula A17, the Fix phenotypes are reversed with S. meliloti NRG185 and NRG34, and there is a correlation between the host specificity and succinoglycan oligosaccharide structure. S. meliloti NRG185 produces oligosaccharides that are almost fully succinylated, with two succinate groups per subunit, whereas the oligosaccharides produced by S. meliloti Rm41 include many nonsuccinylated subunits, as well as subunits with a single succinate group and others with malate. The results of this study demonstrated the following: (i) incompatibility is not a consequence of an avirulence factor or lack of Nod factor activity; (ii) the Fix+ phenotypes are succinoglycan dependent; (iii) there is structural variability in the succinoglycan oligosaccharide populations between S. meliloti strains; (iv) the structural nature of the succinoglycan oligosaccharides is correlated to compatibility; most importantly, (v) an S. meliloti Rm41 derivative, carrying exo genes from an M. truncatula A17-compatible strain, produced a modified population of succinoglycan oligosaccharides (similar to the donor strain) and was Fix+ on A17.

scholarly journals Activity of Sinorhizobium meliloti NodAB and NodH Enzymes on Thiochitooligosaccharides

2002 ◽  
Vol 184 (14) ◽  
pp. 4039-4043 ◽  
Author(s):  
Audrey M. Southwick ◽  
Lai-Xi Wang ◽  
Sharon R. Long ◽  
Yuan C. Lee
Keyword(s):  
Sinorhizobium Meliloti ◽  
Signal Molecules ◽  
Gene Products ◽  
Nod Factors ◽  
Nod Factor ◽  
Glycosidic Bonds

ABSTRACT Rhizobium bacteria synthesize signal molecules called Nod factors that elicit responses in the legume root during nodulation. Nod factors, modified N-acylated β-(1,4)-N-acetylglucosamine, are synthesized by the nodulation (nod) gene products. We tested the ability of three Sinorhizobium meliloti nod gene products to modify Nod factor analogs with thio linkages instead of O-glycosidic bonds in the oligosaccharide backbone.

scholarly journals Sinorhizobium fredii HH103 Invades Lotus burttii by Crack Entry in a Nod Factor–and Surface Polysaccharide–Dependent Manner

2016 ◽  
Vol 29 (12) ◽  
pp. 925-937 ◽  
Author(s):  
Sebastián Acosta-Jurado ◽  
Dulce-Nombre Rodríguez-Navarro ◽  
Yasuyuki Kawaharada ◽  
Juan Fernández Perea ◽  
Antonio Gil-Serrano ◽  
...  
Keyword(s):  
Root Hairs ◽  
Broad Host Range ◽  
Capsular Polysaccharides ◽  
Dependent Manner ◽  
Nod Factors ◽  
Sinorhizobium Fredii ◽  
Nod Factor ◽  
Mesorhizobium Loti ◽  
Infection Threads ◽  
Surface Polysaccharide

Sinorhizobium fredii HH103-Rifr, a broad host range rhizobial strain, induces nitrogen-fixing nodules in Lotus burttii but ineffective nodules in L. japonicus. Confocal microscopy studies showed that Mesorhizobium loti MAFF303099 and S. fredii HH103-Rifr invade L. burttii roots through infection threads or epidermal cracks, respectively. Infection threads in root hairs were not observed in L. burttii plants inoculated with S. fredii HH103-Rifr. A S. fredii HH103-Rifr nodA mutant failed to nodulate L. burttii, demonstrating that Nod factors are strictly necessary for this crack-entry mode, and a noeL mutant was also severely impaired in L. burttii nodulation, indicating that the presence of fucosyl residues in the Nod factor is symbiotically relevant. However, significant symbiotic impacts due to the absence of methylation or to acetylation of the fucosyl residue were not detected. In contrast S. fredii HH103-Rifr mutants showing lipopolysaccharide alterations had reduced symbiotic capacity, while mutants affected in production of either exopolysaccharides, capsular polysaccharides, or both were not impaired in nodulation. Mutants unable to produce cyclic glucans and purine or pyrimidine auxotrophic mutants formed ineffective nodules with L. burttii. Flagellin-dependent bacterial mobility was not required for crack infection, since HH103-Rifr fla mutants nodulated L. burttii. None of the S. fredii HH103-Rifr surface-polysaccharide mutants gained effective nodulation with L. japonicus.

scholarly journals Improved Characterization of Nod Factors and Genetically Based Variation in LysM Receptor Domains Identify Amino Acids Expendable for Nod Factor Recognition in Lotus spp.

2010 ◽  
Vol 23 (1) ◽  
pp. 58-66 ◽  
Author(s):  
Anita S. Bek ◽  
Jørgen Sauer ◽  
Mikkel B. Thygesen ◽  
Jens Ø. Duus ◽  
Bent O. Petersen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Developmental Stages ◽  
Lotus Japonicus ◽  
Resonance Spectroscopy ◽  
Nod Factors ◽  
Nod Factor ◽  
Mesorhizobium Loti ◽  
Flight Mass Spectrometry ◽  
Bradyrhizobium Sp ◽  
Ultrahigh Performance Liquid Chromatography

Formation of functional nodules is a complex process depending on host–microsymbiont compatibility in all developmental stages. This report uses the contrasting symbiotic phenotypes of Lotus japonicus and L. pedunculatus, inoculated with Mesorhizobium loti or the Bradyrhizobium sp. (Lotus), to investigate the role of Nod factor structure and Nod factor receptors (NFR) for rhizobial recognition, infection thread progression, and bacterial persistence within nodule cells. A key contribution was the use of 800 MHz nuclear magnetic resonance spectroscopy and ultrahigh-performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry for Nod factor analysis. The Nod factor decorations at the nonreducing end differ between Bradyrhizobium sp. (Lotus) and M. loti, and the NFR1/NFR5 extracellular regions of L. pedunculatus and L. japonicus were found to vary in amino acid composition. Genetic transformation experiments using chimeric and wild-type receptors showed that both receptor variants recognize the structurally different Nod factors but the later symbiotic phenotype remained unchanged. These results highlight the importance of additional checkpoints during nitrogen-fixing symbiosis and define several amino acids in the LysM domains as expendable for perception of the two differentially carbamoylated Nod factors.

scholarly journals N-Deacetylation of Sinorhizobium meliloti Nod Factors Increases Their Stability in the Medicago sativa Rhizosphere and Decreases Their Biological Activity

2000 ◽  
Vol 13 (1) ◽  
pp. 72-79 ◽  
Author(s):  
Christian Staehelin ◽  
Michael Schultze ◽  
Ken Tokuyasu ◽  
Véréna Poinsot ◽  
Jean-Claude Promé ◽  
...  
Keyword(s):  
Biological Activity ◽  
Medicago Sativa ◽  
Sinorhizobium Meliloti ◽  
Colletotrichum Lindemuthianum ◽  
Signal Molecules ◽  
Glycosyl Hydrolases ◽  
Nod Factors ◽  
Nod Factor ◽  
Chemical Structures

Nod factors excreted by rhizobia are signal molecules that consist of a chitin oligomer backbone linked with a fatty acid at the nonreducing end. Modifications of the Nod factor structures influence their stability in the rhizosphere and their biological activity. To test the function of N-acetyl groups in Nod factors, NodSm-IV(C16:2,S) from Sinorhizobium meliloti was enzymatically N-deacetylated in vitro with purified chitin deacetylase from Colletotrichum lindemuthianum. A family of partially and completely deacetylated derivatives was produced and purified. The most abundant chemical structures identified by mass spectrometry were GlcN(C16:2)-GlcNAc-GlcNH2-GlcNAc(OH)(S), GlcN(C16:2)-GlcNAc-GlcNH2-GlcNH2(OH)(S), and GlcN(C16:2)-GlcNH2-GlcNH2-GlcNH2(OH)(S). In contrast to NodSm-IV(C16:2,S), the purified N-deacetylated derivatives were stable in the rhizosphere of Medicago sativa, indicating that the N-acetyl groups make the carbohydrate moiety of Nod factors accessible for glycosyl hydrolases of the host plant. The N-deacetylated derivatives displayed only a low level of activity in inducing root hair deformation. Furthermore, the N-deacetylated molecules were not able to stimulate Nod factor degradation by M. sativa roots, a response elicited by active Nod factors. These data show that N-acetyl groups of Nod factors are required for biological activity.

scholarly journals Nodulation Gene Mutants of Mesorhizobium loti R7A—nodZ and nolL Mutants Have Host-Specific Phenotypes on Lotus spp.

2009 ◽  
Vol 22 (12) ◽  
pp. 1546-1554 ◽  
Author(s):  
Patsarin Rodpothong ◽  
John T. Sullivan ◽  
Kriangsak Songsrirote ◽  
David Sumpton ◽  
Kenneth W. J.-T. Cheung ◽  
...  
Keyword(s):  
Factor Structure ◽  
Infection Thread ◽  
Plant Responses ◽  
Nod Factors ◽  
Nod Factor ◽  
Mesorhizobium Loti ◽  
Terminal Residue ◽  
Nodulation Genes ◽  
Thread Formation ◽  
Infection Thread Formation

Rhizobial Nod factors induce plant responses and facilitate bacterial infection, leading to the development of nitrogen-fixing root nodules on host legumes. Nodule initiation is highly dependent on Nod-factor structure and, hence, on at least some of the nodulation genes that encode Nod-factor production. Here, we report the effects of mutations in Mesorhizobium loti R7A nodulation genes on nodulation of four Lotus spp. and on Nod-factor structure. Most mutants, including a ΔnodSΔnolO double mutant that produced Nod factors lacking the carbamoyl and possibly N-methyl groups on the nonreducing terminal residue, were unaffected for nodulation. R7AΔnodZ and R7AΔnolL mutants that produced Nod factors without the (acetyl)fucose on the reducing terminal residue had a host-specific phenotype, forming mainly uninfected nodule primordia on Lotus filicaulis and L. corniculatus and effective nodules with a delay on L. japonicus. The mutants also showed significantly reduced infection thread formation and Nin gene induction. In planta complementation experiments further suggested that the acetylfucose was important for balanced signaling in response to Nod factor by the L. japonicus NFR1/NFR5 receptors. Overall the results reveal differences in the sensitivity of plant perception with respect to signaling leading to root hair deformation and nodule primordium development versus infection thread formation and rhizobial entry.

scholarly journals MtENOD20, a Nod Factor-Inducible Molecular Marker for Root Cortical Cell Activation

1999 ◽  
Vol 12 (7) ◽  
pp. 604-614 ◽  
Author(s):  
Vanessa Vernoud ◽  
Etienne-Pascal Journet ◽  
David G. Barker
Keyword(s):  
Transcriptional Activation ◽  
Sinorhizobium Meliloti ◽  
Cell Activation ◽  
Cortical Cell ◽  
Structural Parameter ◽  
Nod Factors ◽  
Root Cortex ◽  
Nod Factor ◽  
Transgenic Alfalfa ◽  
Infection Threads

The spatio-temporal expression pattern of the Medicago truncatula ENOD20 gene during early stages of nodulation has been analyzed with transgenic alfalfa (M. varia) expressing a pMtENOD20-GUS chimeric fusion. Our results show that transcriptional activation of this gene occurs initially in dividing inner cortical cells corresponding to sites of nodule primordium formation and subsequently in root hairs containing infection threads. Use of Sinorhizobium meliloti nod gene mutants that uncouple nodule organogenesis from infection has confirmed that early MtENOD20 transcription is tightly linked to cortical cell activation (CCA). Furthermore, these experiments have revealed that an S. meliloti nodH mutant, defective in Nod factor sulfation and epidermal cell activation, is nevertheless able to elicit low-level CCA. Purified S. meliloti Nod factors trigger MtENOD20 transcription very rapidly (within 12 to 24 h) in the root cortex, and studies with transgenic alfalfa show that Nod factors are able to elicit gene expression coupled to CCA at concentrations as low as 10-11 M. Finally, we have made use of a range of synthetic lipo-chitooligosaccharides to show that fatty acid chain length is an important structural parameter in the capacity of Nod factors to elicit CCA. Taken together, our results suggest that pMtENOD20-GUS transgenic lines should prove valuable tools in future studies of Rhizobium and Nod factor-elicited CCA.

scholarly journals Potentiometric Surfactant Sensor Based on 1,3-Dihexadecyl-1H-benzo[d]imidazol-3-ium for Anionic Surfactants in Detergents and Household Care Products

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3627
Author(s):  
Nikola Sakač ◽  
Dubravka Madunić-Čačić ◽  
Dean Marković ◽  
Lucija Hok ◽  
Robert Vianello ◽  
...  
Keyword(s):  
Anionic Surfactants ◽  
Negative Impact ◽  
Ion Pair ◽  
Two Phase ◽  
Potentiometric Response ◽  
Titration Curves ◽  
Signal Change ◽  
Nernstian Slope ◽  
Good Agreement

A 1,3-dihexadecyl-1H-benzo[d]imidazol-3-ium-tetraphenylborate (DHBI-TPB) ion-pair implemented in DHBI-TPB surfactant sensor was used for the potentiometric quantification of anionic surfactants in detergents and commercial household care products. The DHBI-TPB ion-pair was characterized by FTIR spectroscopy and computational analysis which revealed a crucial contribution of the C–H∙∙∙π contacts for the optimal complex formation. The DHBI-TPB sensor potentiometric response showed excellent analytical properties and Nernstian slope for SDS (60.1 mV/decade) with LOD 3.2 × 10−7 M; and DBS (58.4 mV/decade) with LOD 6.1 × 10−7 M was obtained. The sensor possesses exceptional resistance to different organic and inorganic interferences in broad pH (2–10) range. DMIC used as a titrant demonstrated superior analytical performances for potentiometric titrations of SDS, compared to other tested cationic surfactants (DMIC > CTAB > CPC > Hyamine 1622). The combination of DHBI-TPB sensor and DMIC was successfully employed to perform titrations of the highly soluble alkane sulfonate homologues. Nonionic surfactants (increased concentration and number of EO groups) had a negative impact on anionic surfactant titration curves and a signal change. The DHBI-TPB sensor was effectively employed for the determination of technical grade anionic surfactants presenting the recoveries from 99.5 to 101.3%. The sensor was applied on twelve powered samples as well as liquid-gel and handwashing home care detergents containing anionic surfactants. The obtained results showed good agreement compared to the outcomes measured by ISE surfactant sensor and a two-phase titration method. The developed DHBI-TPB surfactant sensor could be used for quality control in industry and has great potential in environmental monitoring.

scholarly journals Crystal structure of thermostable alkylsulfatase SdsAP from Pseudomonas sp. S9

2017 ◽  
Vol 37 (3) ◽  
Author(s):  
Lifang Sun ◽  
Pu Chen ◽  
Yintao Su ◽  
Zhixiong Cai ◽  
Lingwei Ruan ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Sodium Dodecyl ◽  
Titration Calorimetry ◽  
Pseudomonas Sp ◽  
Sterol Carrier Protein ◽  
Dimerization Domain ◽  
Alkyl Chains ◽  
Hydrolysis Of

A novel alkylsulfatase from bacterium Pseudomonas sp. S9 (SdsAP) was identified as a thermostable alkylsulfatases (type III), which could hydrolyze the primary alkyl sulfate such as sodium dodecyl sulfate (SDS). Thus, it has a potential application of SDS biodegradation. The crystal structure of SdsAP has been solved to a resolution of 1.76 Å and reveals that SdsAP contains the characteristic metallo-β-lactamase-like fold domain, dimerization domain, and C-terminal sterol carrier protein type 2 (SCP-2)-like fold domain. Kinetic characterization of SdsAP to SDS by isothermal titration calorimetry (ITC) and enzymatic activity assays of constructed mutants demonstrate that Y246 and G263 are important residues for its preference for the hydrolysis of ‘primary alkyl’ chains, confirming that SdsAP is a primary alkylsulfatase.

scholarly journals The Removal of Methylene Blue Dye from Aqueous Solutions Using Activated and Non-Activated Bentonites

2003 ◽  
Vol 21 (5) ◽  
pp. 451-462 ◽  
Author(s):  
Sameer Al-Asheh ◽  
Fawzi Banat ◽  
Leena Abu-Aitah
Keyword(s):  
Methylene Blue ◽  
Ph Value ◽  
Sodium Dodecyl ◽  
Freundlich Isotherm ◽  
Blue Dye ◽  
Batch Adsorption ◽  
Ionic Surfactant ◽  
Methylene Blue Dye ◽  
X Ray Diffraction ◽  
Initial Ph

An improvement in the adsorption capacity of naturally available bentonite towards water pollutants such as Methylene Blue dye (MBD) is certainly needed. For this purpose, sodium bentonite was activated by two methods: (1) treatment with sodium dodecyl sulphate (SDS) as an ionic surfactant and (2) thermal treatment in an oven operated at 850°C. Batch adsorption tests were carried out on removing MBD from aqueous solution using the above-mentioned bentonites. It was found that the effectiveness of bentonites towards MBD removal was in the following order: thermal-bentonite > SDS-bentonite > natural bentonite. X-Ray diffraction analysis showed that an increase in the microscopic bentonite platelets on treatment with SDS was the reason behind the higher uptake of MBD. An increase in sorbent concentration or initial pH value of the solutions resulted in a greater removal of MBD from the solution. An increase in temperature led to an increase in MBD uptake by the bentonites studied in this work. The Freundlich isotherm model was employed and found to represent the experimental data well.

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