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Gas concentrations and isotopic signatures can unveil microbial metabolisms and their responses to environmental changes in soil. Currently, few methods measure in situ soil trace gases such as the products of nitrogen and carbon cycling or volatile organic compounds (VOCs) that constrain microbial biochemical processes like nitrification, methanogenesis, respiration, and microbial communication. Versatile trace gas sampling systems that integrate soil probes with sensitive trace gas analyzers could fill this gap with in situ soil gas measurements that resolve spatial (centimeters) and temporal (minutes) patterns. We developed a system that integrates new porous and hydrophobic sintered polytetrafluoroethylene (sPTFE) diffusive soil gas probes that non-disruptively collect soil gas samples with a transfer system to direct gas from multiple probes to one or more central gas analyzer(s) such as laser and mass spectrometers. Here, we demonstrate the feasibility and versatility of this automated multiprobe system for soil gas measurements of isotopic ratios of nitrous oxide (δ18 O, δ15 N, and the 15 N site preference of N 2 O), methane, carbon dioxide (δ13 C), and VOCs. First, we used an inert silica matrix to challenge probe measurements under controlled gas conditions. By changing and controlling system flow parameters, including the probe flow rate, we optimized recovery of representative soil gas samples while reducing sampling artifacts on subsurface concentrations. Second, we used this system to provide a real-time window into the impact of environmental manipulation of irrigation and soil redox conditions on in situ N 2 O and VOC concentrations. Moreover, to reveal the dynamics in the stable isotope ratios of N 2 O (i.e., 14 N 14 N 16 O, 14 N 15 N 16 O, 15 N 14 N 16 O, and 14 N 14 N 18 O), we developed a new high-precision laser spectrometer with a reduced sample volume demand. Our integrated system – a tunable infrared laser direct absorption spectrometry (TILDAS) in parallel with Vocus proton transfer reaction mass spectrometry (PTR-MS), in line with sPTFE soil gas probes – successfully quantified isotopic signatures for N 2 O, CO 2 , and VOCs in real time as responses to changes in the dry–wetting cycle and redox conditions. Broadening the collection of trace gases that can be monitored in the subsurface is critical for monitoring biogeochemical cycles, ecosystem health, and management practices at scales relevant to the soil system. [ABSTRACT FROM AUTHOR]

Soil microbes vigorously produce and consume gases that reflect active soil biogeochemical processes. Soil gas measurements are therefore a powerful tool to monitor microbial activity. Yet, the majority of soil gases lack non-disruptive subsurface measurement methods at spatiotemporal scales relevant to microbial processes and soil structure. To address this need, we developed a soil gas sampling system that uses novel diffusive soil probes and sample transfer approaches for high-resolution sampling from discrete subsurface regions. Probe sampling requires transferring soil gas samples to above-ground gas analyzers where concentrations and isotopologues are measured. Obtaining representative soil gas samples has historically required balancing disruption to soil gas composition with measurement frequency and analyzer volume demand. These considerations have limited attempts to quantify trace gas spatial concentration gradients and heterogeneity at scales relevant to the soil microbiome. Here, we describe our new flexible diffusive probe sampling system integrated with a modified, reduced volume trace gas analyzer and demonstrate its application for subsurface monitoring of biogeochemical cycling of nitrous oxide (N2O) and its site-specific isotopologues, methane, carbon dioxide, and nitric oxide in controlled soil columns. The sampling system observed reproducible responses of soil gas concentrations to manipulations of soil nutrients and redox state, providing a new window into the microbial response to these key environmental forcings. Using site-specific N2O isotopologues as indicators of microbial processes, we constrain the dynamics of in situ microbial activity. Unlocking trace gas messengers of microbial activity will complement -omics approaches, challenge subsurface models, and improve understanding of soil heterogeneity to disentangle interactive processes in the subsurface biome. [ABSTRACT FROM AUTHOR]

Gas concentrations and isotopic signatures can unveil microbial metabolisms and their responses to environmental changes in soil. Currently, few methods measure soil trace gases such as the products of nitrogen and carbon cycling, or volatile organic compounds (VOCs), that could constrain microbial biochemical processes like nitrification, methanogenesis, respiration, and microbial communication. Versatile trace gas sampling systems that integrate soil probes with sensitive trace gas analyzers could fill this gap with measurements resolving spatial (centimeter scale) and temporal (minutes) variations in concentrations and isotopic signatures of in situ soil gases. We developed a system that integrates new 15 cm long sintered PTFE diffusive soil gas probes with various infrared spectrometers and a VOC mass spectrometer. The system is based on porous and hydrophobic soil probes that non-disruptively collect and transport gas from multiple probes to one or more central gas analyzers. Here, we demonstrate the feasibility and versatility of an automated multi-probe system for soil gas measurements of isotopic ratios of nitrous oxide (d18O, d15N, and the 15N site-preference of N2O), methane, carbon dioxide (d13C), and VOCs. First, we used an inert silica matrix to challenge probe measurements under controlled gas conditions. By changing and controlling system flow parameters, including probe flow rate, we optimized recovery of representative soil gas samples while reducing sampling artifacts on subsurface concentrations. Second, we forced environmental manipulations in soil-filled columns to demonstrate real time detection of subsurface gas dynamics in response to irrigation and soil redox conditions. In addition, we developed a new laser spectrometer to recover isotope ratios for 14N14N16O ("d446"), 14N15N16O ("d456"), 15N14N16O ("d546"), and 14N14N18O ("d448") with high precision and low concentration dependence. We captured temporal subsurface gas pulses in CO2, N2O, and VOCs. This demonstrated the potential for diffusive-based probes to couple to trace gas sensors for soil health and fertility studies, and to inform high-throughput meta-omics, leading to the development of a suite of powerful new tools for soil analysis. [ABSTRACT FROM AUTHOR]