Branched glycerol dialkyl glycerol tetraethers (brGDGTs) are ubiquitous and well preserved sedimentary biomarkers. These compounds serve as important palaeoenvironmental indicators due to strong empirical correlations between brGDGT distributions and temperature and pH in modern environments. However, the mechanistic link between temperature, pH, and brGDGT production has been impossible to ascertain thus far due to the absence of a clear biological source for brGDGTs. Here, we report that oxygen limitation triggers brGDGT production in at least one cultured species of Acidobacteria and confirm for the first time the biosynthesis of three structural varieties of brGDGTs, including an uncharacterised isomer of brGDGT Ic. This discovery helps explain why brGDGT producers have been so difficult to identify and provides a pathway towards uncovering the genetic basis and biological function of brGDGTs, which will lead to a more comprehensive understanding of their palaeoenvironmental significance. If the oxygen effects observed here apply more broadly, the empirical calibrations for brGDGT-based temperature and pH reconstructions may currently be missing the effects of oxygen as a relevant and possibly dominant control in the environmental distributions of brGDGTs.

Branched glycerol dialkyl glycerol tetraethers (brGDGTs) are bacterial membrane lipids that are frequently employed as paleoenvironmental proxies because of the strong empirical correlations between their relative abundances and environmental temperature and pH. Despite the ubiquity of brGDGTs in modern and paleoenvironments, the source organisms of these enigmatic compounds have remained elusive, requiring paleoenvironmental applications to rely solely on observed environmental correlations. Previous laboratory and environmental studies have suggested that the globally abundant bacterial phylum of the Acidobacteria may be an important brGDGT producer in nature. Here, we report on experiments with a cultured Acidobacterium, Solibacter usitatus, that makes a large portion of its cellular membrane (24 ± 9% across all experiments) out of a structurally diverse set of tetraethers including the common brGDGTs Ia, IIa, IIIa, Ib, and IIb. Solibacter usitatus was grown across a range of conditions including temperatures from 15 to 30°C, pH from 5.0 to 6.5, and O2 from 1% to 21%, and demonstrated pronounced shifts in the degree of brGDGT methylation across these growth conditions. The temperature response in culture was in close agreement with trends observed in published environmental datasets, supporting a physiological basis for the empirical relationship between brGDGT methylation number and temperature. However, brGDGT methylation at lower temperatures (15 and 20°C) was modulated by culture pH with higher pH systematically increasing the degree of methylation. In contrast, pH had little effect on brGDGT cyclization, supporting the hypothesis that changes in bacterial community composition may underlie the link between cyclization number and pH observed in environmental samples. Oxygen concentration likewise affected brGDGT methylation highlighting the potential for this environmental parameter to impact paleotemperature reconstruction. Low O2 culture conditions further resulted in the production of uncommon brGDGT isomers that could be indicators of O2 limitation. Finally, the production of brGTGTs (trialkyl tetraethers) in addition to the previously discovered iso‐C15‐based mono‐ and diethers in S. usitatus suggests a potential biosynthetic pathway for brGDGTs that uses homologs of the archaeal tetraether synthase (Tes) enzyme for tetraether synthesis from diethers.

As humans increasingly dominate the nitrogen cycle, deposition of reactive nitrogen (Nr) will continue to have adverse consequences for ecosystems. In the Rocky Mountains, Nr deposition remains elevated and has become increasingly dominated by ammonium, despite efforts to reduce emissions. Currently, spatial models of Nr deposition do not fully account for urban and agricultural emissions, sources that contribute to the observed high rates of ammonium deposition in adjacent ecosystems. To address this gap in the Colorado Front Range, we measured Nr deposition along a transect from urban and agricultural plains to subalpine forests. We found elevated values of wet Nr deposition at the urban and foothill sites (4.7 and 4.4 kg N ha−1 yr−1, respectively), and lower values at the montane and subalpine sites (2.5–2.8 kg N ha−1 yr−1). Ammonium dominated wet and bulk Nr deposition, accounting for approximately 69% of bulk Nr deposition. Seasonally, bulk Nr deposition was highest in the spring months, when air masses from the plains are transported west into the mountains. Previous work has demonstrated that high elevations of the Colorado Front Range are especially sensitive to Nr deposition due to thin soil and minimal vegetation. Our results indicate that despite lower precipitation, the fire‐prone forested foothills receive even greater Nr deposition than higher elevations, due to proximity to urban and agricultural Nr sources. The interaction between elevated Nr deposition and wildfire in this region may pose a risk to water supplies and ecosystems, and is an important topic for future research.