Open AccessPersistence and stability of genetically manipulated derivatives of Enterobacter agglomerans in soil microcosms
Author(s) -
EvguenievaHackenberg Elena,
SelenskaPobell Sonja,
Klingmüller Walter
Publication year - 1994
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1111/j.1574-6941.1994.tb00242.x
Subject(s) - biology , persistence (discontinuity) , microcosm , enterobacter , pantoea agglomerans , microbiology and biotechnology , bacteria , ecology , genetics , escherichia coli , geotechnical engineering , gene , engineering
Molecular methods and conventional plating were applied to monitor Enterobacter agglomerans 339 derivatives carrying a Tn5‐Mob or an npt I‐cassette in unsterile soil microcosms. The plate counts of the introduced bacteria decreased continuously in time until undetectable on selective media. In contrast, hybridization of the total DNA directly isolated from inoculated soil samples showed that the target sequences detected corresponded to a much higher number of bacteria than indicated by plating. By PCR‐amplification and hybridization of the soil DNA we could show that asignificant number of target sequences still persisted in the soil microcosms, even when the inoculated bacteria were not able to make colonies on selective agar plates. The Tn 5 marker caused instabilities in the genome of the bacteria studied. Some of the clones that grew in the soil samples had rearrangements in their genome. The detection of E. agglomerans 339 derivatives carrying the immobile npt I‐cassette was also dependent on its location in the bacterial genome.