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Analysis of Plasmid pJP4 Horizontal Transfer and Its Impact on Bacterial Community Structure in Natural Soil

Journal of microbiology and biotechnology v.15 no.2 , 2005년, pp.376 - 383  
KIM TAE SUNG (School of Agricultural Biotechnology, Seoul National University, Biological Resources Division, Biodiversity Research Department, National Institute of Environmental Research ) ;  KIM MI SOON ( School of Agricultural Biotechnology, Seoul National University ) ;  JUNG MEE KUM ( School of Agricultural Biotechnology, Seoul National University ) ;  JOE MIN JEONG ( School of Agricultural Biotechnology, Seoul National University ) ;  AHN JAE HYUNG ( School of Agricultural Biotechnology, Seoul National University ) ;  OH KYOUNG HEE ( Biological Resources Division, Biodiversity Research Department, National Institute of Environmental Research ) ;  LEE MIN HYO ( Biological Resources Division, Biodiversity Research Department, National Institute of Environmental Research ) ;  KIM MIN KYUN ( School of Agricultural Biotechnology, Seoul National University ) ;  KA JONG OK ( School of Agricultural Biotechnology, Seoul National University)
Abstract

Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

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KIM MI SOON(5) JUNG MEE KUM(3) Ka, Jong-Ok(34)

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