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Modified Organism
Pseudomonas fluorescens strain SBW25 modified for biocontrol of fungal pathogens
Record information and status
Record ID
100725
Status
Published
Date of creation
2009-09-03 20:36 UTC (manoela.miranda@cbd.int)
Date of last update
2012-08-10 20:14 UTC (dina.abdelhakim@cbd.int)
Date of publication
2012-08-10 20:14 UTC (dina.abdelhakim@cbd.int)

Living Modified Organism identity
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.

LMO name
Pseudomonas fluorescens strain SBW25 modified for biocontrol of fungal pathogens
Transformation event
SBW25:tgl
Developer(s)
Swedish University of Agricultural Sciences (SLU), Department of Microbiology
Box 7025
750 07 Uppsala, Sweden
Description
The bacterium, Pseudomonas fluorescens, was chromosomally tagged with genes kilA, telAB conferring resistance to potassium tellurite, the constitutive promoter P-psbA, the gfp gene producing green fluorescent protein and luxAB genes encoding bioluminescence production. All these genes were inserted in order to monitor and trace the bacterium in the wheat plant-soil environment.
Recipient Organism or Parental Organisms
The term Recipient organism refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas Parental organisms refers to those that were involved in cross breeding or cell fusion.
Pseudomonas fluorescens - PSEFL
Characteristics of the transformation process
Vector
pUTtgl
Techniques used for the modification
  • Electroporation
Introduced or modified genetic elements
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
Green Fluorescent Protein gene - Aequorea victoria - Crystal Jellyfish, Water Jellyfish, AEQVI
Selectable marker genes and reporter genes
Thylakoid membrane protein gene promoter - Amaranthus hybridus - Slim Amaranth, Green Amaranth, Pigweed
Luciferase alpha and beta subunit fusion gene - Vibrio harveyi - Vibrio, V. harveyi
Selectable marker genes and reporter genes
kilA gene - Enterobacter aerogenes - Enterobacter
Selectable marker genes and reporter genes
telAB gene - Enterobacter aerogenes - Enterobacter
Selectable marker genes and reporter genes
Notes regarding the genetic elements introduced or modified in this LMO
Intended function of each constituent part of the insert in the GMO:

kilA, telAB: these genes confer resistance to the chemical compound potassium tellurite (K2TeO3). The use of these genes as marker genes was developed to suit strains that were aimed to be released in field trials, where it is not desirable to use antibiotic resistance genes as marker tools for tracking the bacteria. Makes selective plating on potassium tellurite possible.

P-psbA: a constitutive promoter.

GFP: GFP emits green light at 508 nm upon ultraviolet light illumination at 396 nm, no additional substrates needed. GFP is a measurement of the total number of cells since it is expressed regardless of metabolic activity in the cells.

LUXAB: The phenotype, bioluminescence, is dependent on the energy reserves of the cells, FMNH2. Therefore, the luxAB genes can be used to determine the number of metabolically active cells. Bioluminescence requires n-decanal as a substrate.

Information on the inserted DNA sequences
kilA, telAB: The tellurite resistance is encoded by the cryptic telAB and kilA genes of plasmid RK2 from Klebsiella aerogenes. PpsbA/gfp/luxAB: The PpsbA/gfp/luxAB genes were excised from plasmid pUTgfplux as a NotI fragment and ligated into the unique NotI-site of pUTtel.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
Common use(s)
  • Biocontrol

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