Evaluation of Toxicity of Cercospora piaropi in a Mycoherbicide Formulation by Using Bacterial Bioluminescence and the Ames Mutagenicity Tests

AbstractUsing the lux operon (luxCDABE) of bacterial bioluminescence system as an autonomous luminous reporter has been demonstrated in bacteria, plant and mammalian cells. However, applications of bacterial bioluminescence-based imaging have been limited because of its low brightness. Here, we engineered the bacterial luciferase (heterodimer of luxA and luxB) by fusion with Venus, a bright variant of yellow fluorescent protein, to induce bioluminescence resonance energy transfer (BRET). By using decanal as an externally added substrate, color change and ten-times enhancement of brightness was achieved in Escherichia coli when circularly permuted Venus was fused to the C-terminus of luxB. Expression of the Venus-fused luciferase in human embryonic kidney cell lines (HEK293T) or in Nicotiana benthamiana leaves together with the substrate biosynthesis-related genes (luxC, luxD and luxE) enhanced the autonomous bioluminescence. We believe the improved luciferase will forge the way towards the potential development of autobioluminescent reporter system allowing spatiotemporal imaging in live cells.

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Induced polypeptide synthesis during the development of bacterial bioluminescence.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)33522-6 ◽

1976 ◽

Vol 251 (9) ◽

pp. 2541-2549 ◽

Cited By ~ 5

Author(s):

G A Michaliszyn ◽

E A Meighen

Keyword(s):

Bacterial Bioluminescence ◽

Polypeptide Synthesis

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A sensitive assay for flavin mononucleotide using the bacterial bioluminescence reaction

Biochemical Medicine ◽

10.1016/0006-2944(67)90011-7 ◽

1967 ◽

Vol 1 (3) ◽

pp. 252-260 ◽

Cited By ~ 14

Author(s):

Emmett W. Chappelle ◽

Grace Lee Picciolo ◽

Robert H. Altland

Keyword(s):

Flavin Mononucleotide ◽

Sensitive Assay ◽

Bacterial Bioluminescence

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Bacterial bioluminescence: organization, regulation, and application of the lux genes

The FASEB Journal ◽

10.1096/fasebj.7.11.8370470 ◽

1993 ◽

Vol 7 (11) ◽

pp. 1016-1022 ◽

Cited By ~ 179

Author(s):

Edward A. Meighen

Keyword(s):

Bacterial Bioluminescence ◽

Lux Genes

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The use of bioluminescence as a reporter to study the adherence of the plant growth promoting rhizopseudomonads 7NSK2 and ANP15 to canola roots

Canadian Journal of Microbiology ◽

10.1139/m93-046 ◽

1993 ◽

Vol 39 (3) ◽

pp. 329-334 ◽

Cited By ~ 15

Author(s):

J. Boelens ◽

D. Zoutman ◽

J. Campbell ◽

W. Verstraete ◽

W. Paranchych

Keyword(s):

Plant Growth ◽

Freundlich Isotherm ◽

Root Surface ◽

Broad Host Range ◽

Bacterial Bioluminescence ◽

Range Vector ◽

Plant Growth Promoting ◽

Specific Receptors ◽

Growth Promoting ◽

Kinetics Of

The adherence of the plant growth promoting rhizopseudomonads Pseudomonas aeruginosa 7NSK2 and Pseudomonas fluorescens ANP15 to canola roots (Brassica campestris L. c.v. Tobin) was examined by means of a bacterial bioluminescence system. The bioluminescence broad host range vector pDLUX-I was constructed from pLAFR-I and the lux A–E genes of Vibrio fischerii. This vector was conjugally transferred into the plant growth promoting rhizopseudomonads 7NSK2 and ANP15. The transformed strains were constitutively bioluminescent at an optimal temperature of 21 °C. The measured bioluminescence was directly proportional to the density of the bacteria in suspension and was the same for both planktonic and sessile bacteria adhering to the root surface. The adherence of the plant growth promoting rhizopseudomonads was proportional to the density of the bacterial inoculum, approached saturation at 60 min, and was reversible. The kinetics of the microbial adhesion was described by a Freundlich isotherm suggesting that the adherence of the bacteria to the canola root surface does not involve specific receptors. We conclude that the pDLUX-I vector is an easy and accurate way to study the kinetics of microbial adherence to the rhizoplane.Key words: rhizopseudomonads, bioluminescence, adhesion, plant growth promotion.not available

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