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Appl Environ Microbiol, July 1998, p. 2634-2638, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Effect of Insertion Site and Metabolic Load on the Environmental Fitness of a Genetically Modified Pseudomonas fluorescens Isolate

Frans A. A. M. De Leij,1 Catherine E. Thomas,1 Mark J. Bailey,2 John M. Whipps,3 and James M. Lynch1,*

School of Biological Sciences, University of Surrey, Guildford, Surrey GU2 5XH,1 Natural Environment Research Council, Institute of Virology and Environmental Microbiology, Oxford OX 3SR,2 and Horticulture Research International, Wellesbourne, Warwick CV35 9EF,3 United Kingdom

Received 20 May 1997/Accepted 5 May 1998

An isolate of Pseudomonas fluorescens (SBW25) was modified with different marker genes (lacZY, aph-1, and xylE). These marker genes were inserted singly or in combination into two separate (1 Mbp apart) and presumably nonessential sites (-6- and Ee) on the chromosome of SBW25. This allowed the production of a range of genetically modified SBW25 variants that differed with respect to insertion site of the marker genes and metabolic burden. The environmental fitness of the different SBW25 variants was tested in soil, in the rhizosphere of wheat and pea, and on the phylloplane of wheat. Reduced environmental fitness of the different variants was mainly attributed to the extra metabolic burden of novel gene expression, whereas choice of insertion site was of little significance. Changes in environmental fitness were dependent on the environmental conditions; an environment, such as soil, with a low microbial carrying capacity had a negative effect on the environmental fitness of variants with a large metabolic load. In environments with a larger carrying capacity, such as the rhizosphere of pea, environmental fitness of variants with a large metabolic load was not significantly different from that of variants with a smaller metabolic burden.

* Corresponding author. Mailing address: School of Biological Sciences, University of Surrey, Guildford, Surrey GU2 5XH, United Kingdom. Phone: 00 44 1483 259721. Fax: 00 44 1483 259728. E-mail: j.lynch{at}surrey.ac.uk.

Appl Environ Microbiol, July 1998, p. 2634-2638, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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