Microbial Biomass, Growth, and CUE Estimation

Soil microbial total DNA-C concentration was used as a proxy for biomass C; DNA extraction was carried out on a soil aliquot of 0.25 g using PowerSoil-htp 96-well soil DNA isolation kit following the manufacturer's instructions (MO BIO Laboratories, UK). Another set of identical DNA extraction was performed following addition of 25 µL of DO¹³C solution and overnight (16 h) incubation in the dark at field moisture capacity. Both extracts were analyzed in the size-exclusion chromatographic (SEC) mode on a liquid chromatography isotope ratio mass spectrometer LC-IRMS (HPLC system coupled to a Delta + XP IRMS through an LC IsoLink interface; Thermo Fisher Scientific, Germany)^{46 (link)}. This allowed us to obtain the DNA-C content and the proportion of DO¹³C in microbial DNA from soils with and without substrate addition. Microbial CUE was estimated as DNA-¹³C/(DNA-¹³C + ∑CO₂-¹³C), where ∑CO₂-¹³C is the cumulative DO¹³C lost during respiration. Microbial turnover rate (synonymously referred to as growth rate) was calculated as DNA-¹³C/DNA-C.

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Malik A.A., Puissant J., Buckeridge K.M., Goodall T., Jehmlich N., Chowdhury S., Gweon H.S., Peyton J.M., Mason K.E., van Agtmaal M., Blaud A., Clark I.M., Whitaker J., Pywell R.F., Ostle N., Gleixner G, & Griffiths R.I. (2018). Land use driven change in soil pH affects microbial carbon cycling processes. Nature Communications, 9, 3591.

Publication 2018

PowerSoil-htp 96-well soil DNA isolation kit Delta + XP IRMS LC IsoLink interface

Hplc Isolation Isotope Liquid chromatography Respiration Size exclusion chromatographic

Corresponding Organization : UK Centre for Ecology & Hydrology

Other organizations : Lancaster University, Helmholtz Centre for Environmental Research, Max Planck Institute for Biogeochemistry, Imperial College London, Rothamsted Research

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Variable analysis

independent variables

Addition of 25 µL of DO^13C solution and overnight (16 h) incubation in the dark at field moisture capacity

dependent variables

Soil microbial total DNA-C concentration (used as a proxy for biomass C)
Proportion of DO^13C in microbial DNA
Microbial carbon use efficiency (CUE) estimated as DNA-^13C/(DNA-^13C + ∑CO2-^13C)
Microbial turnover rate (growth rate) calculated as DNA-^13C/DNA-C

control variables

Soil aliquot of 0.25 g
DNA extraction using PowerSoil-htp 96-well soil DNA isolation kit following the manufacturer's instructions

controls

Positive control: Identical DNA extraction performed with addition of 25 µL of DO^13C solution and overnight (16 h) incubation in the dark at field moisture capacity
Negative control: Identical DNA extraction performed without addition of DO^13C solution

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