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Genetic basis of evolutionary adaptation by Escherichia coli to stressful cycles of freezing, thawing and growth.

ABSTRACT: Microbial evolution experiments offer a powerful approach for coupling changes in complex phenotypes, including fitness and its components, with specific mutations. Here we investigate mutations substituted in 15 lines of Escherichia coli that evolved for 1000 generations under freeze-thaw-growth (FTG) conditions. To investigate the genetic basis of their improvements, we screened many of the lines for mutations involving insertion sequence (IS) elements and identified two genes where multiple lines had similar mutations. Three lines had IS150 insertions in cls, which encodes cardiolipin synthase, and 8 lines had IS150 insertions in the uspA-uspB intergenic region, encoding two universal stress proteins. Another line had an 11-bp deletion mutation in the cls gene. Strain reconstructions and competitions demonstrated that this deletion is beneficial under the FTG regime in its evolved genetic background. Further experiments showed that this cls mutation helps maintain membrane fluidity after freezing and thawing and improves freeze-thaw (FT) survival. Reconstruction of isogenic strains also showed that the IS150 insertions in uspA/B are beneficial under the FTG regime. The evolved insertions reduce uspB transcription and increase both FT survival and recovery, but the physiological mechanism for this fitness improvement remains unknown.

SUBMITTER: Sleight SC

PROVIDER: S-EPMC2535694 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature

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Genetic basis of evolutionary adaptation by Escherichia coli to stressful cycles of freezing, thawing and growth.

Sleight Sean C SC Orlic Christian C Schneider Dominique D Lenski Richard E RE

Genetics 20080830 1

Microbial evolution experiments offer a powerful approach for coupling changes in complex phenotypes, including fitness and its components, with specific mutations. Here we investigate mutations substituted in 15 lines of Escherichia coli that evolved for 1000 generations under freeze-thaw-growth (FTG) conditions. To investigate the genetic basis of their improvements, we screened many of the lines for mutations involving insertion sequence (IS) elements and identified two genes where multiple l ...[more]

PMID: 18757947

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Project description:Lucerne (Medicago sativa L.) is a major component of the crops used in dry-land farming systems in China and its management is associated with notable nitrous oxide (N2O) emissions. A high proportion of these emissions is more likely to occur during periods when the soil undergoes freezing and thawing cycles. In this study, the effects of freeze/thaw cycles on N2O emissions and related factors were investigated in lucerne grasslands. The hypothesis was tested whether increased emissions resulted from a disruption of nitrification or denitrification caused by variations in soil temperatures and water contents. Three days (3 × 24 h) were chosen, where conditions represented freezing and thawing cycles. N2O emissions were measured for a fallow control (F) and two grasslands where lucerne had been cultivated for 4 and 11 years. Soil temperature, soil water content, soil microbial biomass carbon (MBC), soil microbial biomass nitrogen (MBN), soil ammonium nitrogen (NH4 +-N), and soil nitrate nitrogen (NO3 --N) contents were measured. Moreover, the quantities of soil nitrification and denitrification microbes were assessed. Variations in N2O emissions were strongly affected by freeze/thaw cycles, and emissions of 0.0287 ± 0.0009, 0.0230 ± 0.0019, and 0.3522 ± 0.0029 mg m-2 h-1 were found for fallow, 4-year-old, and 11-year-old grasslands, respectively. Pearson correlation analyses indicated that N2O emissions were significantly correlated with the soil water content, temperature, NH4 +-N content, and the number of nitrosobacteria and denitrifying bacteria at a soil depth of 0-100 mm. The numbers of nitrosobacteria and denitrifying bacteria correlated significantly with soil temperature at this soil depth. MBN and soil NH4 +-N contents correlated significantly with soil water content at this depth. Principal component analysis highlighted the positive effects of the number of denitrifying bacteria on N2O emissions during the freeze/thaw period. Furthermore, soil temperature and the number of nitrosobacteria at the tested soil depth (0-100 mm) also played a significant role. This shows that soil freeze/thaw cycles strongly impacted both N2O emissions and the diurnal range, and the number of denitrifying bacteria was mainly influenced by soil temperature and soil NH4 +-N content. The number of denitrifying bacteria was the dominant variable affecting N2O emissions from lucerne grasslands during the assessed soil freeze/thaw period on the Loess Plateau, China.

Dataset Information

Genetic basis of evolutionary adaptation by Escherichia coli to stressful cycles of freezing, thawing and growth.

Publications

Genetic basis of evolutionary adaptation by Escherichia coli to stressful cycles of freezing, thawing and growth.

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