Survival of a lacZY-marked strain of Pseudomonas corrugata following a field release.

Choi, HY; Ryder, MH; Gillings, MR; Stokes, HW; Ophel-Keller, KM; Veal, DA

Survival of a lacZY-marked strain of Pseudomonas corrugata following a field release.

Choi, HY Ryder, MH Gillings, MR Stokes, HW Ophel-Keller, KM Veal, DA

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Publication Type:: Journal Article
Citation:: FEMS Microbiol Ecol, 2003, 43 (3), pp. 367 - 374
Issue Date:: 2003-04-01

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Field	Value	Language
dc.contributor.author	Choi, HY	en_US
dc.contributor.author	Ryder, MH	en_US
dc.contributor.author	Gillings, MR	en_US
dc.contributor.author	Stokes, HW	en_US
dc.contributor.author	Ophel-Keller, KM	en_US
dc.contributor.author	Veal, DA	en_US
dc.date.issued	2003-04-01	en_US
dc.identifier.citation	FEMS Microbiol Ecol, 2003, 43 (3), pp. 367 - 374	en_US
dc.identifier.uri	http://hdl.handle.net/10453/8583
dc.description.abstract	Abstract Pseudomonas corrugata, strain 2140, a biological control agent of take-all disease of wheat, was originally isolated from an acidic red-brown earth soil in New South Wales, Australia. A spontaneous rifampicin-resistant mutant of this bacterium was marked with the disarmed transposon, Tn7::lacZY. This marked strain (2140RlacZY) was introduced into a calcareous sandy loam soil (pH 8) in South Australia. Up to 4 years after its release, P. corrugata 2140RlacZY cells were re-isolated, single colony purified and stored at -80 degrees C. Re-isolated bacteria, including re-isolates obtained 3 (22 re-isolates) and 4 (3 re-isolates) years after release, were examined for stability of the lacZY insert site and for gross chromosomal changes. Hybridization of a cloned lacZY fragment to DNA extracted from the soil re-isolates did not reveal any major changes to the lacZY insert site. Gross chromosomal changes were further examined by restriction endonuclease fingerprinting and PCR based on repetitive sequences (repetitive extragenic palindromic-, enterobacterial repetitive intergeneric consensus- and BOX-PCR). MspI digests distinguished the lacZY-marked strain from the parental strain. None of the genetic techniques used revealed any polymorphisms between the original 2140RlacZY-marked strain and the soil re-isolates. The results demonstrated that the chromosomal landscape within and around the insertion site of the lacZY construct had not altered in the re-isolated bacteria during the 4 years the organism had been in the field.	en_US
dc.language	eng	en_US
dc.relation.ispartof	FEMS Microbiol Ecol	en_US
dc.relation.isbasedon	10.1111/j.1574-6941.2003.tb01077.x	en_US
dc.subject.classification	Microbiology	en_US
dc.title	Survival of a lacZY-marked strain of Pseudomonas corrugata following a field release.	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	43	en_US
utslib.location.activity	England	en_US
utslib.for	060504 Microbial Ecology	en_US
utslib.for	05 Environmental Sciences	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
dc.location.activity	ISI:000181663600009	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	closed_access
pubs.issue	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	43	en_US

Abstract:

Abstract Pseudomonas corrugata, strain 2140, a biological control agent of take-all disease of wheat, was originally isolated from an acidic red-brown earth soil in New South Wales, Australia. A spontaneous rifampicin-resistant mutant of this bacterium was marked with the disarmed transposon, Tn7::lacZY. This marked strain (2140RlacZY) was introduced into a calcareous sandy loam soil (pH 8) in South Australia. Up to 4 years after its release, P. corrugata 2140RlacZY cells were re-isolated, single colony purified and stored at -80 degrees C. Re-isolated bacteria, including re-isolates obtained 3 (22 re-isolates) and 4 (3 re-isolates) years after release, were examined for stability of the lacZY insert site and for gross chromosomal changes. Hybridization of a cloned lacZY fragment to DNA extracted from the soil re-isolates did not reveal any major changes to the lacZY insert site. Gross chromosomal changes were further examined by restriction endonuclease fingerprinting and PCR based on repetitive sequences (repetitive extragenic palindromic-, enterobacterial repetitive intergeneric consensus- and BOX-PCR). MspI digests distinguished the lacZY-marked strain from the parental strain. None of the genetic techniques used revealed any polymorphisms between the original 2140RlacZY-marked strain and the soil re-isolates. The results demonstrated that the chromosomal landscape within and around the insertion site of the lacZY construct had not altered in the re-isolated bacteria during the 4 years the organism had been in the field.

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http://hdl.handle.net/10453/8583