Induction and cultivation of a stable L‐form of <i>Bacillus subtilis</i>

Allan, Eunice J.

doi:10.1111/j.1365-2672.1991.tb02946.x

Cited by 31 publications

(33 citation statements)

References 10 publications

(8 reference statements)

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“…Identifying such conditions and their link with eisosome regulation represents a key future challenge. We note that many unexpected aspects of bacterial cell biology have emerged from so-called L-form growth, which entails removal of the cell wall and osmotic stabilization (Allan, 1991;Errington, 2013;Mercier et al, 2014). Similar insights into yeast cell biology might be uncovered from continued work on protoplasts and conditions that weaken the cell wall.…”

Section: Resultsmentioning

confidence: 96%

Eisosomes provide membrane reservoirs for rapid expansion of the yeast plasma membrane

Kabeche

Howard

Moseley

2015

Journal of Cell Science

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Cell surface area rapidly increases during mechanical and hypoosmotic stresses. Such expansion of the plasma membrane requires 'membrane reservoirs' that provide surface area and buffer membrane tension, but the sources of this membrane remain poorly understood. In principle, the flattening of invaginations and buds within the plasma membrane could provide this additional surface area, as recently shown for caveolae in animal cells. Here, we used microfluidics to study the rapid expansion of the yeast plasma membrane in protoplasts, which lack the rigid cell wall. To survive hypoosmotic stress, yeast cell protoplasts required eisosomes, protein-based structures that generate long invaginations at the plasma membrane. Both budding yeast and fission yeast protoplasts lacking eisosomes were unable to expand like wild-type protoplasts during hypoosmotic stress, and subsequently lysed. By performing quantitative fluorescence microscopy on single protoplasts, we also found that eisosomes disassembled as surface area increased. During this process, invaginations generated by eisosomes at the plasma membrane became flattened, as visualized by scanning electron microscopy. We propose that eisosomes serve as tensiondependent membrane reservoirs for expansion of yeast cells in an analogous manner to caveolae in animal cells.

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Section: Resultsmentioning

confidence: 96%

Eisosomes provide membrane reservoirs for rapid expansion of the yeast plasma membrane

Kabeche

Howard

Moseley

2015

Journal of Cell Science

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“…It is known that B. subtilis can grow and proliferate after a permanent loss of the cell wall as stable nonreverting L-forms (1,72). Recently, a defined B. subtilis strain was constructed that can easily be converted from rodshaped to L-form cells (19,34).…”

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confidence: 99%

Cell Envelope Stress Response in Cell Wall-Deficient L-Forms of Bacillus subtilis

Wolf

Domı́nguez-Cuevas

Daniel

et al. 2012

Antimicrob Agents Chemother

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b L-forms are cell wall-deficient bacteria that can grow and proliferate in osmotically stabilizing media. Recently, a strain of the Gram-positive model bacterium Bacillus subtilis was constructed that allowed controlled switching between rod-shaped wildtype cells and corresponding L-forms. Both states can be stably maintained under suitable culture conditions. Because of the absence of a cell wall, L-forms are known to be insensitive to ␤-lactam antibiotics, but reports on the susceptibility of L-forms to other antibiotics that interfere with membrane-anchored steps of cell wall biosynthesis are sparse, conflicting, and strongly influenced by strain background and method of L-form generation. Here we investigated the response of B. subtilis to the presence of cell envelope antibiotics, with regard to both antibiotic resistance and the induction of the known LiaRS-and BceRS-dependent cell envelope stress biosensors. Our results show that B. subtilis L-forms are resistant to antibiotics that interfere with the bactoprenol cycle, such as bacitracin, vancomycin, and mersacidin, but are hypersensitive to nisin and daptomycin, which both affect membrane integrity. Moreover, we established a lacZ-based reporter gene assay for L-forms and provide evidence that LiaRS senses its inducers indirectly (damage sensing), while the Bce module detects its inducers directly (drug sensing).

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“…Stable L‐forms were cultivated for 24 h at 25°C and 60 r.p.m. in L‐Phase Broth (LPB) (Allan 1991). L‐forms were harvested from 20 ml aliquots (2500 g for 10 min, MSE Chilspin, Sanyo Gallenkamp, Leicester, UK) and re‐suspended in 5% (w/v) mannitol.…”

Section: Methodsmentioning

confidence: 99%

The symbiosis of Bacillus subtilis L-forms with Chinese cabbage seedlings inhibits conidial germination of Botrytis cinerea

Walker

Ferguson

Booth

et al. 2002

Lett Appl Microbiol

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Aims: To establish whether germination of Botrytis cinerea was affected by the symbiosis of Bacillus subtilis L-form bacteria with Chinese cabbage. Methods and Results: Germinating seeds of Chinese cabbage were co-cultivated with either L-forms of Bacillus subtilis or 5% (w/v) mannitol by soaking for 3 h. Seeds were then washed in sterile water, sown on a minimal medium and incubated in controlled conditions. L-form symbiosis was detected over a time course by ELISA. Conidial germination of Botrytis cinerea was signi®cantly reduced on cotyledonous leaves of L-form-treated plants compared with controls. Conclusions: Symbiosis of B. subtilis L-form bacteria during seed germination of Chinese cabbage inhibits conidial germination in plants on subsequent exposure to Botrytis cinerea. Signi®cance and Impact of the Study: This is the ®rst account of plant symbiosis with L-form bacteria showing antagonism to a fungal plant pathogen. This has promising implications for the use of this L-form as a biocontrol agent.

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Induction and cultivation of a stable L‐form of Bacillus subtilis

Cited by 31 publications

References 10 publications

Eisosomes provide membrane reservoirs for rapid expansion of the yeast plasma membrane

Eisosomes provide membrane reservoirs for rapid expansion of the yeast plasma membrane

Cell Envelope Stress Response in Cell Wall-Deficient L-Forms of Bacillus subtilis

The symbiosis of Bacillus subtilis L-forms with Chinese cabbage seedlings inhibits conidial germination of Botrytis cinerea

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