Abstract
Heavy water (H 2 18 O) has been used to label DNA of soil microorganisms in stable isotope probing experiments, yet no measurements have been reported for the 18O content of DNA from soil incubated with heavy water. Here we present the first measurements of atom% 18O for DNA extracted from soil incubated with the addition of H 2 18 O. Four experiments were conducted to test how the atom% 18O of DNA, extracted from Ponderosa Pine forest soil incubated with heavy water, was affected by the following variables: (1) time, (2) nutrients, (3) soil moisture, and (4) atom% 18O of added H2O. In the time series experiment, the atom% 18O of DNA increased linearly (R 2 = 0.994, p < 0.01) over the first 72 h of incubation. In the nutrient addition experiment, there was a positive correlation (R 2 = 0.991, p = 0.006) between the log10 of the amount of tryptic soy broth, a complex nutrient broth, added to soil and the log10 of the atom% 18O of DNA. For the experiment where soil moisture was manipulated, the atom% 18O of DNA increased with higher soil moisture until soil moisture reached 30%, above which 18O enrichment of DNA declined as soils became more saturated. When the atom% 18O for H2O added was varied, there was a positive linear relationship between the atom% 18O of the added water and the atom% 18O of the DNA. Results indicate that quantification of 18O incorporated into DNA from H 2 18 O has potential to be used as a proxy for microbial growth in soil.
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Acknowledgments
This work was supported by the Office of Science (BER), US Department of Energy, Grant No. DE-FG02-04ER63883 and the National Science Foundation, Grant No. EF-0747397. We gratefully acknowledge Dave Harris’s IRMS expertise at the UC Davis Stable Isotope Facility, and we thank Rebecca Daly for valuable comments on the manuscript.
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Blazewicz, S.J., Schwartz, E. Dynamics of 18O Incorporation from H 2 18O into Soil Microbial DNA. Microb Ecol 61, 911–916 (2011). https://doi.org/10.1007/s00248-011-9826-7
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DOI: https://doi.org/10.1007/s00248-011-9826-7