Activity of antibiotics produced by Bacillus subtilis and Bacillus pumilus against common fungal contaminants of plant tissue cultures

Danby, S; Hampson, S. P.; Joshi, S.; Sigee, David C.; Epton, H. A. S.; Leifert, Carlo

doi:10.1007/978-94-011-0790-7_45

Cited by 6 publications

(4 citation statements)

References 4 publications

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“…Fungal seeded medium was poured into Micro-detection dishes (Proteus Molecular Design Ltd, Marplc, Chcshire, UK). Antibiotic activity assays were performed in Microdetection plates because they allow the detection of lower concentrations of antibiotics than assays carried out with conventional Petri dishes (Danby et al 1994). After the agar had set 4 mm wells were cut into the agar with a cork borer and filled with 30 pl of the cell-free culture filtrates from batch cultures of Bacillus spp.…”

Section: In Vltro Assay For Sntlfungal Sctivltymentioning

confidence: 99%

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Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45

Leifert

Chidburee

et al. 1995

Journal of Applied Bacteriology

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248

129

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Bacillus subtilis CL27 and B. pumilus CL45 showed similar activity against Botrytis cinerea in in vitro plate assays. In a seedling bioassay, however, B. subtilis CL27 had activity similar to a commercial fungicide while B. pumilus CL45 failed completely to prevent seedling damping-off caused by Bot. cinerea. Antibiotic production by the two Bacillus strains was found to depend on the growth substrate and highest antibiotic production was found on media based on homogenized cabbage tissue. Antibiotic activity was found to depend on the pH and nutrient concentration in the assay medium. Antifungal antibiotics produced by B. subtilis CL27 and B. pumilus CL45 in different fermentation media were separated by thin layer chromatography. As suspected from the activity spectrum, three antibiotics (one with activity against Alternaria brassicicola, one with activity against Botrytis cinerea and one with activity against both fungi) could be detected in the fermentation broth of CL27, but only one in the fermentation broth of CL45. The two antibiotics produced by strain CL27 with activity against A. brassicicola were identified as peptides since their bands on the TLC plates developed a green to blue/green colour after treatment with 4,4'-tetramethyldiamino-diphenylmethane (TDM) reagent. The third antibiotics produced by strain CL27 and antibiotic produced by CL45 had a similar Rf-value and appeared not to be peptides based on the reaction with TDM. However, they showed a slightly different activity spectrum when tested against a range of different fungi. Antibiotic production was clearly indicated as the mode of action of in vivo biocontrol by strain CL27 against damping off caused by Bot. cinerea of Astilbe micro-plants, because a u.v.-induced antibiotic negative mutant strain CL27b showed no activity in seedling bioassays in vivo. Also the mutant strain CL27a which produced the two peptide antibiotics but had lost the ability to produce the non-peptide antibiotic, showed greatly reduced in vivo activity.

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Section: In Vltro Assay For Sntlfungal Sctivltymentioning

confidence: 99%

“…pumilus CL45 (two strains with similar in vitro activity against Botrytis cinerea) and two mutant strains of CL27 with either no or reduced antibiotic production. Strains CL27 and CI,45 have previously been shown to have a different range of activity when tested against various plant pathogenic fungi (Danby et al 1994).…”

mentioning

confidence: 99%

Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45

Leifert

Chidburee

et al. 1995

Journal of Applied Bacteriology

Self Cite

248

129

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show abstract

“…Microdetection plates were designed to allow a two-way diffusion of antimicrobial substances applied to the plates in comparison to conventional Petri dishes (which allow a three-way diffusion) and it was anticipated that this \\auld increase the sensitivity of detection (Danby et al 1994). Once the relative sensitivities of these bioassay s !…”

Section: Ps Antimicrohic[~ Against the Growth Of The Pathogen On Potatomentioning

confidence: 99%

Bioassay methods for the detection of antifungal activity by Pseudomonas antimicrobica against the grey mould pathogen Botrytis cinerea

Walker

Emslie

Allan

1996

Journal of Applied Bacteriology

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Antagonism against the grey mould pathogen Botrytis cinerea by Pseudomonas antimicrobica was demonstrated in vitro and in vivo. Cell-free filtrates showed activity against B. cinerea growing on Potato Dextrose Agar (PDA) in a media-dependent manner with the most distinct antagonism being produced in Czapek Dox Broth (CDB). Cell-free filtrates of CDB-grown cultures also significantly reduced conidial germination of B. cinerea. An assay based on the inhibition of conidial germination was compared with two assays measuring the antagonism of mycelial growth on PDA. The conidial germination bioassay was more sensitive in the detection of this antifungal activity than the Petri dish bioassay while a bioassay using Microdetection plates did not detect antagonism due to the small loading capacity of the latter. The conidial germination bioassay was modified for detection of antibiosis on the surface of strawberry leaves. Significant reductions in percentage conidial germination were recorded on the surface of leaves of both micropropagated and glasshouse grown strawberry plants when the antifungal compounds of Ps. antimicrobica were applied to the leaf tissue with the conidia. In addition, antifungal compounds were also detectable when conidia were applied to leaf tissue which had previously been sprayed with cells of Ps. antimicrobica. These tests indicate that Ps. antimicrobica would be a suitable biocontrol agent for the control of B. cinerea.

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“…Dextrose Agar in Petri dishes and Microdetection plates and against conidial germination on glass slides as described by Edwards (1993). Microdetection plates were designed to allow a two-way diffusion of antimicrobial substances applied to the plates in comparison to conventional Petri dishes (which allow a three-way diffusion) and it was anticipated that this \\auld increase the sensitivity of detection (Danby et al 1994). Once the relative sensitivities of these bioassay s !…”

Section: Ps Antimicrohic[~ Against the Growth Of The Pathogen On Potatomentioning

confidence: 99%

Bioassay methods for the detection of antifungal activity by Pseudomonas antimicrobica against the grey mould pathogen Botrytis cinerea

Walker¹,

Emslie²,

Allan³

1996

J Appl Microbiol

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Antagonism against the grey mould pathogen Botrytis cinerea by Pseudomonas antimicrobica was demonstrated in vitro and in z$z.o. Cell-free filtrates showed activity against B. cinerea growing on Potato Dextrose Agar (PDA) in a media-dependent manner with the most distinct antagonism being produced in Czapek Dox Broth (CDB). Cell-free filtrates of CDB-grown cultures also significantly reduced conidial germination of B. cinerea. An assay based o n the inhibition of conidial germination was compared with two assays measuring the antagonism of mycelial growth o n PDA. T h e conidial germination bioassay was more sensitive in the detection of this antifungal activity than the Petri dish bioassay while a bioassay using Microdetection plates did not detect antagonism d u e to the small loading capacity of the latter. T h e conidial germination bioassay was modified for detection of antibiosis on the surface of strawberry leaves. Significant reductions in percentage conidial germination were recorded on the surface of leaves of both micropropagated and glasshouse grown strawberry plants when the antifungal compounds of Ps. anrimicrubicri were applied to the leaf tissue with the conidia. In addition, antifungal compounds were also detectable when conidia were applied to leaf tissue which had previously been sprayed with cells of Ps. antimicrobica. These tests indicate that Ps. nntimzcrobica would be a suitable biocontrol agent for the control of B. cinerea.

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Activity of antibiotics produced by Bacillus subtilis and Bacillus pumilus against common fungal contaminants of plant tissue cultures

Cited by 6 publications

References 4 publications

Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45

Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45

Bioassay methods for the detection of antifungal activity by Pseudomonas antimicrobica against the grey mould pathogen Botrytis cinerea

Bioassay methods for the detection of antifungal activity by Pseudomonas antimicrobica against the grey mould pathogen Botrytis cinerea

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