An ELISA for the detection of Bacillus subtilis L-form bacteria confirms their symbiosis in strawberry

Ferguson, C.; Booth, Nuala A.; Allan, Eunice J.

doi:10.1046/j.1472-765x.2000.00834.x

Cited by 31 publications

(39 citation statements)

References 12 publications

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“…B. subtilis had been reported to be able to enter the L-form state in earlier laboratory work [16], as well as in environmental studies of plant–microbe interactions [17]. Richard Daniel, then a postdoc in my laboratory, acquired an environmental L-form isolate of B. subtilis from a laboratory in Aberdeen (that of Eunice Allan; [18]) and began investigating its properties.…”

Section: Applying Modern Molecular Cell Biology Methods To the L-formmentioning

confidence: 99%

Cell wall-deficient, L-form bacteria in the 21st century: a personal perspective

Errington

2017

Biochemical Society Transactions

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The peptidoglycan (PG) cell wall is a defining feature of the bacteria. It emerged very early in evolution and must have contributed significantly to the success of these organisms. The wall features prominently in our thinking about bacterial cell function, and its synthesis involves the action of several dozen proteins that are normally essential for viability. Surprisingly, it turns out to be relatively simple to generate bacterial genetic variants called L-forms that completely lack PG. They grow robustly provided that lack of the cell wall is compensated for by an osmoprotective growth medium. Although their existence has been noted and studied on and off for many decades, it is only recently that modern molecular and cellular methods have been applied to L-forms. We used Bacillus subtilis as an experimental model to understand the molecular basis for the L-form switch. Key findings included the discovery that L-forms use an unusual blebbing, or tubulation and scission mechanism to proliferate. This mechanism is completely independent of the normal FtsZ-based division machinery and seems to require only an increased rate of membrane synthesis, leading to an increased surface area-to-volume ratio. Antibiotics that block cell wall precursor synthesis, such as phosphomycin, efficiently induce the L-form switch without the need for genetic change. The same antibiotics turned out to induce a similar L-form switch in a wide range of bacteria, including Escherichia coli, in which we showed that proliferation was again FtsZ-independent. Aside from further basic science, future work on L-forms is likely to focus on their possible role in chronic or recurrent infections, their use as a model in studies of the origins of life, and possibly, biotechnological applications.

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Section: Applying Modern Molecular Cell Biology Methods To the L-formmentioning

confidence: 99%

Cell wall-deficient, L-form bacteria in the 21st century: a personal perspective

Errington

2017

Biochemical Society Transactions

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“…Currently, comparable results to quantify bacteria within environmental samples can only be achieved by time consuming and complex methods such as fluorescence in situ hybridization (FISH) combined with digital imaging analysis (Daims et al 2001) and ELISA (enzyme linked immunosorbent assay) applications using targetspecific antibodies (Ferguson et al 2000). However, FISH with subsequently microscopic quantification were used only for sludge (Daims et al 2001;Snaidr et al 1997), soil (Chatzinotas et al 1998), sediments (Liobet-Brossa et al 1998) or ectomycorrhizospheres (Mogge et al 2000) and have not yet shown to give quantitative results in plant analysis.…”

Section: E Radicincitans Strain and Bacterial Quantificationmentioning

confidence: 99%

Quantification and Localization of Bacteria in Plant Tissues Using Quantitative Real-Time PCR and Online Emission Fingerprinting

2006

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In order to quantify and localize specific bacterial target genes in plant tissue, this project has generated relevant new insights in the combined application of quantitative real-time PCR in parallel with the in situ PCR + probehybridization and online emission fingerprinting using LSM 510 META. After designing an Enterobacter radicincitans species-specific probe, introduced bacterial cells were monitored in growing plant parts and their colonization behaviour was examined in relation to the native bacterial community. For this purpose, the plant growth-promoting rhizobacterial (PGPR) strain Enterobacter radicincitans was applied to Brassica oleracea plants in increasing inoculum concentrations 10 7 , 10 8 and 10 9 cells per plant. Inoculation of 10 9 E. radicincitans cells per plant to Brassica oleracea leaves and roots resulted in significant increases of root, leaf and tuber growth. Total bacterial cell numbers were estimated using quantitative real-time PCR to be between 10 7 and 10 9 cells g -1 fresh leaf weight and about 10 8 cells g -1 fresh root weight of Brassica oleracea plants. Using quantitative real-time PCR, a significant colonization of Brassica oleracea leaves and roots with E. radicincitans cells was measured. Roots were colonized with a density of 10 7 cells g -1 fresh root weight up to at least 14 days after inoculation. That is equivalent to a proportion of E. radicincitans 16S rDNA-gene copy numbers compared to the total bacterial communities of about 10-16%. Online emission fingerprinting established that the introduced bacteria proliferated on and inside the root and that they colonized the intercellular spaces of the root cortex layer. Hence, E. radicincitans was able to successfully compete with the native bacterial population.

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“…In the last years different technique have been developed such as DNA microarray (Ahn and Walt, 2005; Kim et al , 2005; Cho and Tiedje, 2001), PT-PCR (Malorny et al , 2008; Kramer et al , 2009; Mafu et al , 2009), enzyme-linked immunosorbent assay (ELISA) (Ferguson et al , 2000; Tamminen et al , 2004), surface plasmon resonance (SPR) (Maalouf et al , 2007; Oh et al , 2005; Dudak and Boyaci, 2009), magnetic nanoparticle based immunoassay (Malorny et al , 2008; Gu et al , 2003), various formats of biosensors (Gehring et al , 1998; Liao and Ho, 2009; Lu et al , 2009; Wong et al , 2002) and flow cytometry (Busam et al , 2007; Hahn et al , 2008; Hibi et al , 2007; Qin et al , 2008; Moragues et al , 2004; Ferrari et al , 2004). …”

Section: Introductionmentioning

confidence: 99%

Dual channel detection of ultra low concentration of bacteria in real time by scanning fluorescence correlation spectroscopy

Altamore

Lanzanò

Gratton

2013

Meas. Sci. Technol.

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We describe a novel method to detect very low concentrations of bacteria in water. Our device consists of a portable horizontal geometry small confocal microscope with large pinhole and a holder for cylindrical cuvettes containing the sample. Two motors provide a fast rotational and slow vertical motion of the cuvette so the device looks like a simplified flow cytometer without flow. To achieve high sensitivity the design has two detection channels. Bacteria are stained by two different nucleic acid dyes and excited with two different lasers. Data are analyzed with a correlation filter based on particle passage pattern recognition. The passage of a particle through the illumination volume is compared with a Gaussian pattern in both channels. The width of the Gaussian correlates with the time of passage of the particle so one particle is counted when the algorithm finds a match with a Gaussian in both channels. The concentration of particles in the sample is deduced from the total number of coincident hits and the total volume scanned. This portable setup provides higher sensitivity, low cost and it could have a wide use ranging from clinical applications to pollution monitors and water and air quality control.

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An ELISA for the detection of Bacillus subtilis L-form bacteria confirms their symbiosis in strawberry

Cited by 31 publications

References 12 publications

Cell wall-deficient, L-form bacteria in the 21st century: a personal perspective

Cell wall-deficient, L-form bacteria in the 21st century: a personal perspective

Quantification and Localization of Bacteria in Plant Tissues Using Quantitative Real-Time PCR and Online Emission Fingerprinting

Dual channel detection of ultra low concentration of bacteria in real time by scanning fluorescence correlation spectroscopy

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