Characterization in Pseudomonas putida Cg1 of nahR and its role in bacterial survival in soil

Woojun Park, E. L. Madsen

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Sequencing, RFLP analyses and experiments utilizing a lacZ transcriptional reporter fused to the promoter regions of nahR and nahG in Pseudomonas putida Cg1 confirmed that regulation of naphthalene degradation in both P. putida Cg1 and the type strain, P. putida NCIB 9816-4, is consistent with that of NAH7 from P. putida G7. Two nahR knockout strains (RK1 and Cg1-NAHR from P. putida NCIB 9816-4 and Cg1, respectively) showed a growth defect in the presence of naphthalene as sole carbon and energy source. We hypothesized that nahR influences ecological fitness of bacteria in naphthalene-contaminated soil and tested this hypothesis using both parent and nahR-knockout strains introduced to soil microcosms with and without added naphthalene. After 21 days, loss of cell viability was pronounced in the presence of added naphthalene crystals for nahR mutants of both test bacteria, relative to the wild types. Diminished viable counts were attributed to toxicity. Thus, our data indicated that NahR in P. putida Cg1 is virtually identical to its homologues in other pseudomonads and that nahR is required for resistance to naphthalene toxicity, hence the persistence of bacterial cells in soil with high concentrations of naphthalene.

Original languageEnglish
Pages (from-to)209-216
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume66
Issue number2
DOIs
Publication statusPublished - 2004 Dec 1
Externally publishedYes

Fingerprint

Pseudomonas putida
Naphthalene
Soil
Soils
Toxicity
Bacteria
naphthalene
Genetic Promoter Regions
Restriction Fragment Length Polymorphisms
Cell Survival
Carbon
Cells
Degradation
Defects
Crystals
Growth

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Microbiology
  • Microbiology (medical)

Cite this

Characterization in Pseudomonas putida Cg1 of nahR and its role in bacterial survival in soil. / Park, Woojun; Madsen, E. L.

In: Applied Microbiology and Biotechnology, Vol. 66, No. 2, 01.12.2004, p. 209-216.

Research output: Contribution to journalArticle

@article{3e7cf24c18894414960169d022ee1ce4,
title = "Characterization in Pseudomonas putida Cg1 of nahR and its role in bacterial survival in soil",
abstract = "Sequencing, RFLP analyses and experiments utilizing a lacZ transcriptional reporter fused to the promoter regions of nahR and nahG in Pseudomonas putida Cg1 confirmed that regulation of naphthalene degradation in both P. putida Cg1 and the type strain, P. putida NCIB 9816-4, is consistent with that of NAH7 from P. putida G7. Two nahR knockout strains (RK1 and Cg1-NAHR from P. putida NCIB 9816-4 and Cg1, respectively) showed a growth defect in the presence of naphthalene as sole carbon and energy source. We hypothesized that nahR influences ecological fitness of bacteria in naphthalene-contaminated soil and tested this hypothesis using both parent and nahR-knockout strains introduced to soil microcosms with and without added naphthalene. After 21 days, loss of cell viability was pronounced in the presence of added naphthalene crystals for nahR mutants of both test bacteria, relative to the wild types. Diminished viable counts were attributed to toxicity. Thus, our data indicated that NahR in P. putida Cg1 is virtually identical to its homologues in other pseudomonads and that nahR is required for resistance to naphthalene toxicity, hence the persistence of bacterial cells in soil with high concentrations of naphthalene.",
author = "Woojun Park and Madsen, {E. L.}",
year = "2004",
month = "12",
day = "1",
doi = "10.1007/s00253-004-1630-6",
language = "English",
volume = "66",
pages = "209--216",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "2",

}

TY - JOUR

T1 - Characterization in Pseudomonas putida Cg1 of nahR and its role in bacterial survival in soil

AU - Park, Woojun

AU - Madsen, E. L.

PY - 2004/12/1

Y1 - 2004/12/1

N2 - Sequencing, RFLP analyses and experiments utilizing a lacZ transcriptional reporter fused to the promoter regions of nahR and nahG in Pseudomonas putida Cg1 confirmed that regulation of naphthalene degradation in both P. putida Cg1 and the type strain, P. putida NCIB 9816-4, is consistent with that of NAH7 from P. putida G7. Two nahR knockout strains (RK1 and Cg1-NAHR from P. putida NCIB 9816-4 and Cg1, respectively) showed a growth defect in the presence of naphthalene as sole carbon and energy source. We hypothesized that nahR influences ecological fitness of bacteria in naphthalene-contaminated soil and tested this hypothesis using both parent and nahR-knockout strains introduced to soil microcosms with and without added naphthalene. After 21 days, loss of cell viability was pronounced in the presence of added naphthalene crystals for nahR mutants of both test bacteria, relative to the wild types. Diminished viable counts were attributed to toxicity. Thus, our data indicated that NahR in P. putida Cg1 is virtually identical to its homologues in other pseudomonads and that nahR is required for resistance to naphthalene toxicity, hence the persistence of bacterial cells in soil with high concentrations of naphthalene.

AB - Sequencing, RFLP analyses and experiments utilizing a lacZ transcriptional reporter fused to the promoter regions of nahR and nahG in Pseudomonas putida Cg1 confirmed that regulation of naphthalene degradation in both P. putida Cg1 and the type strain, P. putida NCIB 9816-4, is consistent with that of NAH7 from P. putida G7. Two nahR knockout strains (RK1 and Cg1-NAHR from P. putida NCIB 9816-4 and Cg1, respectively) showed a growth defect in the presence of naphthalene as sole carbon and energy source. We hypothesized that nahR influences ecological fitness of bacteria in naphthalene-contaminated soil and tested this hypothesis using both parent and nahR-knockout strains introduced to soil microcosms with and without added naphthalene. After 21 days, loss of cell viability was pronounced in the presence of added naphthalene crystals for nahR mutants of both test bacteria, relative to the wild types. Diminished viable counts were attributed to toxicity. Thus, our data indicated that NahR in P. putida Cg1 is virtually identical to its homologues in other pseudomonads and that nahR is required for resistance to naphthalene toxicity, hence the persistence of bacterial cells in soil with high concentrations of naphthalene.

UR - http://www.scopus.com/inward/record.url?scp=11244337594&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=11244337594&partnerID=8YFLogxK

U2 - 10.1007/s00253-004-1630-6

DO - 10.1007/s00253-004-1630-6

M3 - Article

C2 - 15278309

AN - SCOPUS:11244337594

VL - 66

SP - 209

EP - 216

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 2

ER -