Use of ELISA with Antiexopolysaccharide Antibodies to Evaluate Wheat-Root Colonization by the Rhizobacterium Paenibacillus polymyxa

Yegorenkova, Irina V.; Tregubova, Kristina V.; Matora, L. Yu.; Burygin, Gennady L.; Игнатов, В. В.

doi:10.1007/s00284-010-9622-5

Cited by 19 publications

(9 citation statements)

References 23 publications

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“…EPS are combinations of monomers and polymers consisting of sugar compounds connected via glyosidic linkages 14,24,25 . EPS length, composition and formation vary considerably 26–28 . The main factors that influence EPS interactions include the charges, polymer functional groups, backbone and chain fitness, and the relative concentrations of the constituents 14,24,25 .…”

Section: Discussionmentioning

confidence: 99%

Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum

Timmusk

Copolovici

et al. 2019

Sci Rep

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Fusarium Head Blight (FHB) caused by Fusarium graminearum pathogens constitutes a major threat to agricultural production because it frequently reduces the yield and quality of the crop. The disease severity is predicted to increase in various regions owing to climate change. Integrated management where biocontrol plays an important role has been suggested in order to fight FHB. P. polymyxa A26 is known to be an effective antagonist against F. graminearum. Deeper understanding of the mode of action of P. polymyxa A26 is needed to develop strategies for its application under natural settings in order to effectively overcome the pathogenic effects. This study aims to re-evaluate a former study and reveal whether compounds other than non-ribosomal antibiotic lipopeptides could be responsible for the antagonistic effect, despite what is often reported. Wheat seedlings were grown to maturity and the spikes infected with the pathogen under greenhouse conditions. The development of FHB infection, quantified via the disease incidence severity and 100-kernel weight, was strongly correlated (r > 0.78, p < 0.01) with the content of the polysaccharide component D-glucuronic acid in the biofilm. Furthermore, while increased inoculum density from 106 to 108 cells/ml did not affect wild type performance, a significant increase was observed with the P. polymyxa mutant deficient in nonribosomal lipopeptide synthesis. Our results show that P. polymyxa A26 biofilm extracellular polysaccharides are capable of antagonizing F. graminearum and that the uronate content of the polysaccharides is of critical importance in the antagonism.

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Section: Discussionmentioning

confidence: 99%

Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum

Timmusk

Copolovici

et al. 2019

Sci Rep

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“…Additionally, in the few cases where inoculated strains are tracked, experimental protocols often require high-priced equipment and/or present technical limitations when studying non-model bacteria. These include immunological techniques such as ELISA and immunofluorescence (Hansen et al 1997; Yegorenkova et al 2010) the use of reporter genes including the β‒galactosidase, β‒glucoranidase, bacterial luciferase and green fluorescent protein genes (Compant et al 2005; Kragelund, Hosbond, and Nybroe 1997; Solanki and Garg 2014), and nucleic acid-based methods like FISH (Watt et al 2006). More recently, microscopy (Hulse 2018; Liu et al 2021), microfluidic devices (Aufrecht et al 2018; Massalha et al 2017; O’Neal, Vo, and Alexandre 2020) and genomic approaches (Kröber et al 2014) have been used.…”

Section: Discussionmentioning

confidence: 99%

Biofilm formation and maize root-colonization of seed-endophytic Bacilli isolated from native maize landraces

Gastélum,

Ángeles,

Arellano-Wattenbarger

et al. 2024

Preprint

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Agricultural microbiology seeks to replace the use of agrochemicals with microbe-based products. Plant growth-promoting bacteria (PGPB) are often selected based on their functionsin vitro, and then, their effect on plant development is tested. However, this approach neglects the study of their survival in soil, root-colonization, and the monitoring of beneficial functions in the rhizosphere. This could explain the recurrent lack of success in the transition from lab tests to field applications of natural isolates from novel habitats. In our recent studies, we found that native maize seeds from traditional agroecosystems carry endophytic bacterial communities that are dominated by members of the class Bacilli. As an approach to grasp their PGP potential, we developed protocols to test maize root-colonization of these natural isolates in 1) a short-term hydroponics assayin vitroand 2) a long-term assay in non-sterile soil pots. Our results show thatin vitrobiofilm formation was only partially associated to short-term colonizationin vitro; furthermore, long-term root-colonization in soil pots was not correlated to thein vitroassays. This work highlights the necessity to incorporate root-colonization assays as part of the research strategies in the search for PGPB natural isolates from unexplored habitats, towards the generation of inoculants with increased success in the field.

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“…Lyophilized chicken, mouse, and rat IgG, were purchased from Jackson ImmunoResearch, USA. Polyclonal rabbit antibodies to these IgG molecules were obtained from blood serum of immunized rabbits as described previously [32]. For preventing a crossreaction between the rabbit anti-mouse antibodies and the closely related rat IgG, the anti-mouse antibodies were purified by immunoaffinity chromatography with rat IgG covalently bound to Sepharose 4B (Amersham Biosciences).…”

Section: Methodsmentioning

confidence: 99%

Multiplexed dot immunoassay using Ag nanocubes, Au/Ag alloy nanoparticles, and Au/Ag nanocages

et al. 2012

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We report the first application of Ag nanocubes, Au/Ag alloy nanoparticles, and Au/Ag nanocages in a multiplexed dot immunoassay. The assay principle is based on the staining of analyte drops on a nitrocellulose membrane strip by using multicolor nanoparticles conjugated with biospecific probing molecules. Nanoparticles were prepared by a galvanic replacement reaction between the Ag atoms of silver nanocubes and Au ions of tetrachloroauric acid. Depending on the Ag/Au conversion ratio, the particle plasmon resonance was tuned from 450 to 700 nm and the suspension color changed from yellow to blue. The particles of yellow, red, and blue suspensions were functionalized with chicken, rat, and mouse immuno gamma globulin (IgG) molecular probes, respectively. The multiplex capability of the assay was illustrated by a proof-of-concept experiment involving simultaneous one-step determination of target molecules (rabbit anti-chicken, anti-rat, and anti-mouse antibodies) with a mixture of fabricated conjugates. Under naked eye examination, no cross-colored spots or nonspecific bioconjugate adsorption were observed, and the low detection limit was about 20 fmol.

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Use of ELISA with Antiexopolysaccharide Antibodies to Evaluate Wheat-Root Colonization by the Rhizobacterium Paenibacillus polymyxa

Cited by 19 publications

References 23 publications

Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum

Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum

Biofilm formation and maize root-colonization of seed-endophytic Bacilli isolated from native maize landraces

Multiplexed dot immunoassay using Ag nanocubes, Au/Ag alloy nanoparticles, and Au/Ag nanocages

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